芪参健脾方对SHR大鼠动脉血压及心肌重塑影响的研究
|
摘要
目的：本实验通过优化芪参健脾方的提取工艺，以及观察芪参健脾方对自发性高血压大鼠（SHR）尾动脉收缩压以及心肌组织内血管紧张素Ⅱ（AngⅡ）、TGF-β1/Smad信号转导途径各因子、基质金属蛋白酶1、2、9（MMP-1，MMP-2，MMP-9）和I，Ⅲ型胶原蛋白（Collagen I，CollagenⅢ）表达的影响，评价其降压效果，并考察芪参健脾方对SHR心肌纤维化，高血压所致的心肌重塑的过程中各信号分子含量表达的影响，探索芪参健脾方调节血管紧张素Ⅱ、TGF-β1/Smad信号转导途径内各因子表达水平的机制；进一步研究芪参健脾方对SHR心肌组织质金属蛋白酶1、2、9和和I，Ⅲ型胶原蛋白表达的影响，探求高血压进程中心肌重塑的改善机制；为临床辨证治疗施以芪参健脾方提供科学的实验数据与理论基础，阐释其降压及心肌保护作用的分子机制。
材料与方法：
以水煎法提取补脾益气汤中的有效部位，以黄芪甲苷含量为指标，采用星点设计-效应面法考察提取次数、溶媒用量、提取时间3个因素对黄芪甲苷含量的影响，优选最佳提取工艺条件。
采用随机对照法将60只24周龄的SHR分为模型组（给予蒸馏水，SHR组）、培哚普利组（培哚普利，0.44mg/kg/d）、培哚普利联合中药组（培哚普利，0.4mg/kg/d+中药中剂量，中加西组）、中药高、中、低剂量组（用药量按照每100g大鼠体质量分别折算为4g，2g，1g），每组10只，以同龄同种系正常血压的京都种大鼠（WKY）10只作为正常组（WKY组），大鼠每日一次灌胃。智能无创血压计测量大鼠初次给药前，首次给药后2h，第2周，4周和6周给药后2h大鼠尾动脉收缩压变化情况，连续治疗6周。
大鼠禁食、自由饮水24h后，各组大鼠称重，10%水合氯醛腹腔麻醉（3mg/kg）。大鼠麻醉后固定于鼠台上，用0.5%碘伏消毒大鼠胸腹部皮肤，沿左侧胸骨旁斜形切口，上界为两前肢后缘线，下界为第五肋间，依次剪开皮肤、浅筋膜和深筋膜，在靠胸骨缘处用止血钳钝性分离第3、4肋间，用专用开胸器撑开3、4肋间暴露心脏，将棉纱放置在肺组织旁，防治损伤肺组织。活体取出心脏，迅速取材，取50mg心肌组织放入EP管中并加入1ml Trizol试剂，-70℃冰箱保存，PCR方法检测心肌AngⅡ、TGF-β1、MMP-1，MMP-2和MMP-9的mRNA表达；另取心肌组织50mg入EP管中，-70℃冰箱保存，western方法检测MMP-1，MMP-2和MMP-9蛋白的表达；剩余心肌组织用4%多聚甲醛固定，进行心肌病理形态学观察及免疫组化方法检测I，Ⅲ型胶原蛋白表达。
采用SPSS17.0统计学软件对数据进行处理，实验数据以均数±标准差(X±s)表示，组间均数比较采用单因素方差分析（ANOVA），P＜0.05为差异有统计学意义。
结果：
1.补脾益气汤的最佳提取工艺
采用星点设计-效应面法优选了补脾益气汤的最佳提取工艺，优选后的理论值和预测值相近，说明所建立的数学模型的预测性较好。
2.芪参健脾方对SHR尾动脉收缩压的影响
统计结果表明，用药前，各组大鼠血压均高于WKY组，P＜0.01；药后2h，各组大鼠收缩压无明显变化；用药2周后，中加西药组、西药组血压明显降低，与模型组相比有统计学意义，P＜0.01，用药4周及6周后，中加西药组、西药组血压持续下降，与模型组相比有统计学意义，P＜0.01；中药高、中、低剂量组血压用药2周后也有一定程度的下降，用药4周及6周后血压下降不明显，与模型组相比无统计学意义。
3.芪参健脾方对SHR心肌组织形态学及Collagen I和CollagenⅢ的表达的影响
WKY组大鼠心肌组织胶原纤维分布比较均匀、纤细，心肌间质无纤维化现象；与WKY组相比，SHR组心肌组织呈典型心肌肥厚改变，心肌细胞排列杂乱无章，被深入的纤维分隔紧裹，细胞间网络增粗变密，心肌纤维走向紊乱，可见溶解断裂，细胞间质纤维化增多。与SHR组相比，培哚普利组、培哚普利联合中药组和中药高剂量组心肌细胞直径明显减小，大小一致，排列整齐，心肌纤维无断裂，纤维增生减少。与SHR模型组比较，培哚普利组、培哚普利联合中药组和中药高剂量组中Collagen I和CollagenⅢ的表达显著下降（P<0.01，P<0.05），中剂量组中CollagenⅢ的表达也明显减少（P<0.05）。
4.芪参健脾方对SHR心肌组织AngⅡ和TGF-β1/Smad信号转导途径中各信号分子含量的影响
与模型组比较，培哚普利组、培哚普利联合中药组和中药高剂量组中TGF-β1、AngII mRNA的表达量显著下降（P<0.05）。与模型组比较，培哚普利组，培哚普利联合中药组，中药高、中、低各剂量组TGF-β1蛋白的表达显著降低（P<0.01，P<0.05），培哚普利组，培哚普利联合中药组，中药高剂量组AngⅡ蛋白和Smad3蛋白的表达量明显低于SHR组（P<0.01，P<0.05），培哚普利组，培哚普利联合中药组Smad2蛋白的表达量明显低于SHR组（P<0.01，P<0.05）。
5.芪参健脾方对SHR心肌组织MMP-1，MMP-2和MMP-9表达的影响
与SHR模型组比较，培哚普利组、培哚普利联合中药组和中药高剂量组中MMP-1，MMP-2和MMP-9的mRNA和蛋白的表达量都显著增加（P<0.01，P<0.05），中药中剂量组MMP-1蛋白的表达也量明显增加（P<0.05）；与正常组比较，SHR组心肌呈典型心肌纤维化改变；与SHR模型组比较，培哚普利联合中药组、培哚普利组和芪参健脾方高剂量组中Collagen I和CollagenⅢ的表达显著下降（P<0.01，P<0.05），中剂量组中CollagenⅢ的表达也明显减少（P<0.05）。
结论：
1.采用星点设计-效应面法可优化补脾益气汤的提取工艺。
2.芪参健脾方联合培哚普利治疗对SHR尾动脉收缩压表现出较好的降压效果，治疗4周达到稳定期。
3.芪参健脾方联合培哚普利及单方高剂量的应用能有效改善SHR心肌病理形态结构，同时显著下调了Collagen I，CollagenⅢ的表达，逆转高血压导致的心肌重塑。
4.芪参健脾方联合培哚普利通过调节SHR心肌组织AngⅡ和TGF-β1/Smad信号转导途径中各信号分子的表达，改善高血压进程中心肌细胞的损伤和功能障碍。
5.芪参健脾方联合培哚普利及单方高剂量的应用，明显促进了MMP-1，MMP-2和MMP-9的表达，从而抑制心肌纤维化，发挥降压及保护心肌组织的功能。
Purpose：This study is to optimize the extraction of Jianpi Yiqi decoctio byusing Central Composite Design/Response Surface Methodology and observe theeffects of Qishen Jianpi decoction on anti-hypertension in the rats withspontaneous hypertension and explore the impacts on protein and mRNA expressionof AngⅡ，TGF-β1，Smad2and Smad3, MMP-1,MMP-2,MMP-9collagen I and collagenⅢ,by which we can value its antihypertensive effect and also investigate thefunction of Qishen Jianpi decoction in the mechanisms prevent the developmentof arterial blood pressure in SHR and inhibit accumulation of cardiac collagenfibers. At the same time, we will also discuss how the Qishen Jianpi decoctionregulates the expression of AngⅡ and influences the content of signalingmolecules which are in the TGF-β1/Smad thoroughfare. Besides, we will futherto research about how the Qishen Jianpi decoction influence the expression ofMMP-1,MMP-2,MMP-9collagenI and collagenⅢof cardiac collagen fibers. Meanwhile,we also explore the mechanisms of inhibiting accumulation of cardiac collagenfibers during the process of hypertension. All the experiments have offered thescientific basis and theories for the use of Qishen Jianpi decoction in theclinical treatment and also explain its molecule process of protecting theMyocardial tissue.
Material and method：Theeffectsofthreeindependentvariables,Extractiontimes, ratio of material to solvent and extracting time were investigated withthe content of Astragaloside IV and Astragalus Polysaccharides. Responsesurfaces were delineated according to the mathematic models, and optimumformulations were selected.
Using the randomized controlled method to divide the6024-week-old SHR intothe model group (giving water,SHR group), perindopril group [perindopril,0.4mg/(Kg·d)], perindopril associated with herbgroup[perindopril0.4mg/(Kg·d)+medium gose of herb], and another3groupsindividually high level, middle level and low level dose of herb(the dose isdecided according to the100g weight which are4g,2g,1g) and the normal groupis10mormal arterial pressure WKY at the same age and the same kind (givingpure water, WKY group). The rats are fed once a day. We will detect the systolicpressure’s change of their caudal artery with BP-98A intelligentsphygmomanometer before and2hours,2weeks,4weeks, and6weeks after givingthem the medicine, and the measuring time is2hours after the medicine giving.It will last6weeks.
The rats will be fed only water for24hours, then these groups of rats willbe weighed after given10%chloral hydrate’s anesthetic. We will fix the ratson the desk after anesthetic and sterilize their belly skin with0.5%Iodophor,then we will cut off their belly skin and widen the thoracic cavity dlong theabdominal cavity to make the heart seeable, and take about50g tissue to putit in the EP canal combined with1ml Trizol in the refrigerator under thetemperature of-70℃, RT-PCR method will be used to detect the mRNA content ofAngⅡ, TGF-β1, MMP-1, MMP-2, MMP-9; Take another50g tissue to put it in theEP canal in the refrigerator under the temperature of-70℃, and the Westernblot method will be used to detect theexpression of AngⅡ, TGF-β1, Smad2, Smad3,MMP-1, MMP-2and MMP-9; The rest of tissue will be put into4%paraformal-dehydeto carry on the observation of pathology images and at the same time detect theexpression of CollagenI and CollagenⅢ.
The data will be checked with one factor analysis of variance using theSPSS17.0software, and LSD method will be used to make comparison in the wayof (X±s), if P＜0.05, it will have significant difference.
Results：
1. Central Composite Design／Response Surface Methodology is successfully usedto optimize the extraction of Jianpi Yiqi decoctio.
All the investigated response variables were found to be highly dependenton the formulation variables. The optimized values of extraction times, ratioof material to solvent and extracting time were2,9and1.5, respectively. Theobserved values agreed well with model predicted values.
2. The influence of Qishen Jianpi decoction on the SHR about the caudal artery’ssystolic pressure.
The statistical results explanation: before giving the medicine, thesystolic pressure of these groups of rats are all higher than the group WKY,P<0.01;4hours after giving them the medicine, there is no obvious change aboutthe systolic pressure; After2weeks, the pressure is obvious getting down forthe perindopril group and the perindopril associated with herb group, andcomparing with the model group, it has the statistical meaning, P<0.01; After4weeks and6weeks, the pressure is continuously getting down and it has thestatistical meaning comparing with the model group, P<0.01. while for the3leveldose of herb groups, the pressure is also getting down at some extent after2weeks of using the medicine, however, after4and6weeks, the pressure is notgetting down clearly, and comparing with the model group, it has no statisticalmeaning.
3. The influence of Qishen Jianpi decoction on the cardiac collagen fibersmorphology and the expression of CollagenI and CollagenⅢ.
The cardiac collagen fibers of WKY group are clear and arrange regularly,interstitial collagen fibers is little; The interstitial fibroblast and collagenfibers are clearly in SHR group. Compared with control group, the shape ofmitochondria is better, interstitial fibroblast and collagen fibers are lessin the perindopril group, the perindopril associated with herb group and thehigh dose of herb group. The protein expression of CollagenI and CollagenⅢ issignificantly decreased in perindopril group, perindopril associated with herbgroup and high level dose group compared with SHR group, P<0.01, P<0.05, andthe expression of CollagenⅢ was significantly decreased in middle level dosegroup,P<0.05.
4. The influence of Qishen Jianpi decoction on the dose of signaling moleculewhich are in the SHR TGF-β1/Smad thoyoughfare.
Comparing with the SHR group, the mRNA content of TGF-β1and AngII of theperindopril group, the perindopril associated with herb group and the high doseof herb group is obviously getting down, P<0.05. Comparing with the SHR group,TGF-β1protein’s expression quantity of the perindopril group, the perindoprilassociated with herb group and the3dose of herb group is clearly decrease,P<0.01，P<0.05; The content of AngII and Smad3protein of the perindopril group,the perindopril associated with herb group and the high dose of herb group ismuch lower than the SHR group, P<0.01，P<0.05; The protein expression of Smad2is decreased in perindopril group， perindopril associated with herb group,P<0.01，P<0.05
5. The influence of Qishen Jianpi decoction on the expression of MMP-1, MMP-2,MMP-9and CollagenI, Ⅲ.
The mRNA and the protein expression of MMP-1, MMP-2and MMP-9wassignificantly increased in perindopril group,perindopril associated with herbgroup and high dose group compared with SHR group, P<0.01，P<0.05, and theexpression of MMP-1was significantly increased in middle dose group, P<0.05;The protein expression of Collagen I and CollagenⅢ was significantly decreasedin perindopril group, perindopril associated with herb group and high dose groupcompared with SHR group, P<0.01, P<0.05, and the expression of CollagenⅢ wassignificantly decreased in middle dose group, P<0.05.
Conclusion：
1. Central Composite Design／Response Surface Methodology is successfully usedto optimize the extraction of Jianpi Yiqi decoctio.
2. The middle level dose of the Qishen Jianpi decoction combined with perindoprilshows the better lowing pressure effect on the caudal artery systolic pressure,after4weeks treatment, it will reach the stable level.
3. The Qishen Jianpi decoction combined with perindopril and the high level dose of Qishen Jianpi decoction can improve SHR cardiac pathology images conformation.By inhibiting the expression of Collagen I and CollagenⅢ so as to reverse theaccumulation of cardiac collagen fibers.
4. By regulating the factors from TGF-β1/Smad signaling pathway and AngII,Qishen Jianpi decoction combined with perindopril could modify the functionobstacle of hearts in the development of arterial blood pressure in SHR.
5. The Qishen Jianpi decoction combined with perindopril and the high level doseof Qishen Jianpi decoction can stimulate the expression of MMP-1, MMP-2and MMP-9to exert its function of lowing blood pressure and protect the hearts.
材料与方法：
以水煎法提取补脾益气汤中的有效部位，以黄芪甲苷含量为指标，采用星点设计-效应面法考察提取次数、溶媒用量、提取时间3个因素对黄芪甲苷含量的影响，优选最佳提取工艺条件。
采用随机对照法将60只24周龄的SHR分为模型组（给予蒸馏水，SHR组）、培哚普利组（培哚普利，0.44mg/kg/d）、培哚普利联合中药组（培哚普利，0.4mg/kg/d+中药中剂量，中加西组）、中药高、中、低剂量组（用药量按照每100g大鼠体质量分别折算为4g，2g，1g），每组10只，以同龄同种系正常血压的京都种大鼠（WKY）10只作为正常组（WKY组），大鼠每日一次灌胃。智能无创血压计测量大鼠初次给药前，首次给药后2h，第2周，4周和6周给药后2h大鼠尾动脉收缩压变化情况，连续治疗6周。
大鼠禁食、自由饮水24h后，各组大鼠称重，10%水合氯醛腹腔麻醉（3mg/kg）。大鼠麻醉后固定于鼠台上，用0.5%碘伏消毒大鼠胸腹部皮肤，沿左侧胸骨旁斜形切口，上界为两前肢后缘线，下界为第五肋间，依次剪开皮肤、浅筋膜和深筋膜，在靠胸骨缘处用止血钳钝性分离第3、4肋间，用专用开胸器撑开3、4肋间暴露心脏，将棉纱放置在肺组织旁，防治损伤肺组织。活体取出心脏，迅速取材，取50mg心肌组织放入EP管中并加入1ml Trizol试剂，-70℃冰箱保存，PCR方法检测心肌AngⅡ、TGF-β1、MMP-1，MMP-2和MMP-9的mRNA表达；另取心肌组织50mg入EP管中，-70℃冰箱保存，western方法检测MMP-1，MMP-2和MMP-9蛋白的表达；剩余心肌组织用4%多聚甲醛固定，进行心肌病理形态学观察及免疫组化方法检测I，Ⅲ型胶原蛋白表达。
采用SPSS17.0统计学软件对数据进行处理，实验数据以均数±标准差(X±s)表示，组间均数比较采用单因素方差分析（ANOVA），P＜0.05为差异有统计学意义。
结果：
1.补脾益气汤的最佳提取工艺
采用星点设计-效应面法优选了补脾益气汤的最佳提取工艺，优选后的理论值和预测值相近，说明所建立的数学模型的预测性较好。
2.芪参健脾方对SHR尾动脉收缩压的影响
统计结果表明，用药前，各组大鼠血压均高于WKY组，P＜0.01；药后2h，各组大鼠收缩压无明显变化；用药2周后，中加西药组、西药组血压明显降低，与模型组相比有统计学意义，P＜0.01，用药4周及6周后，中加西药组、西药组血压持续下降，与模型组相比有统计学意义，P＜0.01；中药高、中、低剂量组血压用药2周后也有一定程度的下降，用药4周及6周后血压下降不明显，与模型组相比无统计学意义。
3.芪参健脾方对SHR心肌组织形态学及Collagen I和CollagenⅢ的表达的影响
WKY组大鼠心肌组织胶原纤维分布比较均匀、纤细，心肌间质无纤维化现象；与WKY组相比，SHR组心肌组织呈典型心肌肥厚改变，心肌细胞排列杂乱无章，被深入的纤维分隔紧裹，细胞间网络增粗变密，心肌纤维走向紊乱，可见溶解断裂，细胞间质纤维化增多。与SHR组相比，培哚普利组、培哚普利联合中药组和中药高剂量组心肌细胞直径明显减小，大小一致，排列整齐，心肌纤维无断裂，纤维增生减少。与SHR模型组比较，培哚普利组、培哚普利联合中药组和中药高剂量组中Collagen I和CollagenⅢ的表达显著下降（P<0.01，P<0.05），中剂量组中CollagenⅢ的表达也明显减少（P<0.05）。
4.芪参健脾方对SHR心肌组织AngⅡ和TGF-β1/Smad信号转导途径中各信号分子含量的影响
与模型组比较，培哚普利组、培哚普利联合中药组和中药高剂量组中TGF-β1、AngII mRNA的表达量显著下降（P<0.05）。与模型组比较，培哚普利组，培哚普利联合中药组，中药高、中、低各剂量组TGF-β1蛋白的表达显著降低（P<0.01，P<0.05），培哚普利组，培哚普利联合中药组，中药高剂量组AngⅡ蛋白和Smad3蛋白的表达量明显低于SHR组（P<0.01，P<0.05），培哚普利组，培哚普利联合中药组Smad2蛋白的表达量明显低于SHR组（P<0.01，P<0.05）。
5.芪参健脾方对SHR心肌组织MMP-1，MMP-2和MMP-9表达的影响
与SHR模型组比较，培哚普利组、培哚普利联合中药组和中药高剂量组中MMP-1，MMP-2和MMP-9的mRNA和蛋白的表达量都显著增加（P<0.01，P<0.05），中药中剂量组MMP-1蛋白的表达也量明显增加（P<0.05）；与正常组比较，SHR组心肌呈典型心肌纤维化改变；与SHR模型组比较，培哚普利联合中药组、培哚普利组和芪参健脾方高剂量组中Collagen I和CollagenⅢ的表达显著下降（P<0.01，P<0.05），中剂量组中CollagenⅢ的表达也明显减少（P<0.05）。
结论：
1.采用星点设计-效应面法可优化补脾益气汤的提取工艺。
2.芪参健脾方联合培哚普利治疗对SHR尾动脉收缩压表现出较好的降压效果，治疗4周达到稳定期。
3.芪参健脾方联合培哚普利及单方高剂量的应用能有效改善SHR心肌病理形态结构，同时显著下调了Collagen I，CollagenⅢ的表达，逆转高血压导致的心肌重塑。
4.芪参健脾方联合培哚普利通过调节SHR心肌组织AngⅡ和TGF-β1/Smad信号转导途径中各信号分子的表达，改善高血压进程中心肌细胞的损伤和功能障碍。
5.芪参健脾方联合培哚普利及单方高剂量的应用，明显促进了MMP-1，MMP-2和MMP-9的表达，从而抑制心肌纤维化，发挥降压及保护心肌组织的功能。
Purpose：This study is to optimize the extraction of Jianpi Yiqi decoctio byusing Central Composite Design/Response Surface Methodology and observe theeffects of Qishen Jianpi decoction on anti-hypertension in the rats withspontaneous hypertension and explore the impacts on protein and mRNA expressionof AngⅡ，TGF-β1，Smad2and Smad3, MMP-1,MMP-2,MMP-9collagen I and collagenⅢ,by which we can value its antihypertensive effect and also investigate thefunction of Qishen Jianpi decoction in the mechanisms prevent the developmentof arterial blood pressure in SHR and inhibit accumulation of cardiac collagenfibers. At the same time, we will also discuss how the Qishen Jianpi decoctionregulates the expression of AngⅡ and influences the content of signalingmolecules which are in the TGF-β1/Smad thoroughfare. Besides, we will futherto research about how the Qishen Jianpi decoction influence the expression ofMMP-1,MMP-2,MMP-9collagenI and collagenⅢof cardiac collagen fibers. Meanwhile,we also explore the mechanisms of inhibiting accumulation of cardiac collagenfibers during the process of hypertension. All the experiments have offered thescientific basis and theories for the use of Qishen Jianpi decoction in theclinical treatment and also explain its molecule process of protecting theMyocardial tissue.
Material and method：Theeffectsofthreeindependentvariables,Extractiontimes, ratio of material to solvent and extracting time were investigated withthe content of Astragaloside IV and Astragalus Polysaccharides. Responsesurfaces were delineated according to the mathematic models, and optimumformulations were selected.
Using the randomized controlled method to divide the6024-week-old SHR intothe model group (giving water,SHR group), perindopril group [perindopril,0.4mg/(Kg·d)], perindopril associated with herbgroup[perindopril0.4mg/(Kg·d)+medium gose of herb], and another3groupsindividually high level, middle level and low level dose of herb(the dose isdecided according to the100g weight which are4g,2g,1g) and the normal groupis10mormal arterial pressure WKY at the same age and the same kind (givingpure water, WKY group). The rats are fed once a day. We will detect the systolicpressure’s change of their caudal artery with BP-98A intelligentsphygmomanometer before and2hours,2weeks,4weeks, and6weeks after givingthem the medicine, and the measuring time is2hours after the medicine giving.It will last6weeks.
The rats will be fed only water for24hours, then these groups of rats willbe weighed after given10%chloral hydrate’s anesthetic. We will fix the ratson the desk after anesthetic and sterilize their belly skin with0.5%Iodophor,then we will cut off their belly skin and widen the thoracic cavity dlong theabdominal cavity to make the heart seeable, and take about50g tissue to putit in the EP canal combined with1ml Trizol in the refrigerator under thetemperature of-70℃, RT-PCR method will be used to detect the mRNA content ofAngⅡ, TGF-β1, MMP-1, MMP-2, MMP-9; Take another50g tissue to put it in theEP canal in the refrigerator under the temperature of-70℃, and the Westernblot method will be used to detect theexpression of AngⅡ, TGF-β1, Smad2, Smad3,MMP-1, MMP-2and MMP-9; The rest of tissue will be put into4%paraformal-dehydeto carry on the observation of pathology images and at the same time detect theexpression of CollagenI and CollagenⅢ.
The data will be checked with one factor analysis of variance using theSPSS17.0software, and LSD method will be used to make comparison in the wayof (X±s), if P＜0.05, it will have significant difference.
Results：
1. Central Composite Design／Response Surface Methodology is successfully usedto optimize the extraction of Jianpi Yiqi decoctio.
All the investigated response variables were found to be highly dependenton the formulation variables. The optimized values of extraction times, ratioof material to solvent and extracting time were2,9and1.5, respectively. Theobserved values agreed well with model predicted values.
2. The influence of Qishen Jianpi decoction on the SHR about the caudal artery’ssystolic pressure.
The statistical results explanation: before giving the medicine, thesystolic pressure of these groups of rats are all higher than the group WKY,P<0.01;4hours after giving them the medicine, there is no obvious change aboutthe systolic pressure; After2weeks, the pressure is obvious getting down forthe perindopril group and the perindopril associated with herb group, andcomparing with the model group, it has the statistical meaning, P<0.01; After4weeks and6weeks, the pressure is continuously getting down and it has thestatistical meaning comparing with the model group, P<0.01. while for the3leveldose of herb groups, the pressure is also getting down at some extent after2weeks of using the medicine, however, after4and6weeks, the pressure is notgetting down clearly, and comparing with the model group, it has no statisticalmeaning.
3. The influence of Qishen Jianpi decoction on the cardiac collagen fibersmorphology and the expression of CollagenI and CollagenⅢ.
The cardiac collagen fibers of WKY group are clear and arrange regularly,interstitial collagen fibers is little; The interstitial fibroblast and collagenfibers are clearly in SHR group. Compared with control group, the shape ofmitochondria is better, interstitial fibroblast and collagen fibers are lessin the perindopril group, the perindopril associated with herb group and thehigh dose of herb group. The protein expression of CollagenI and CollagenⅢ issignificantly decreased in perindopril group, perindopril associated with herbgroup and high level dose group compared with SHR group, P<0.01, P<0.05, andthe expression of CollagenⅢ was significantly decreased in middle level dosegroup,P<0.05.
4. The influence of Qishen Jianpi decoction on the dose of signaling moleculewhich are in the SHR TGF-β1/Smad thoyoughfare.
Comparing with the SHR group, the mRNA content of TGF-β1and AngII of theperindopril group, the perindopril associated with herb group and the high doseof herb group is obviously getting down, P<0.05. Comparing with the SHR group,TGF-β1protein’s expression quantity of the perindopril group, the perindoprilassociated with herb group and the3dose of herb group is clearly decrease,P<0.01，P<0.05; The content of AngII and Smad3protein of the perindopril group,the perindopril associated with herb group and the high dose of herb group ismuch lower than the SHR group, P<0.01，P<0.05; The protein expression of Smad2is decreased in perindopril group， perindopril associated with herb group,P<0.01，P<0.05
5. The influence of Qishen Jianpi decoction on the expression of MMP-1, MMP-2,MMP-9and CollagenI, Ⅲ.
The mRNA and the protein expression of MMP-1, MMP-2and MMP-9wassignificantly increased in perindopril group,perindopril associated with herbgroup and high dose group compared with SHR group, P<0.01，P<0.05, and theexpression of MMP-1was significantly increased in middle dose group, P<0.05;The protein expression of Collagen I and CollagenⅢ was significantly decreasedin perindopril group, perindopril associated with herb group and high dose groupcompared with SHR group, P<0.01, P<0.05, and the expression of CollagenⅢ wassignificantly decreased in middle dose group, P<0.05.
Conclusion：
1. Central Composite Design／Response Surface Methodology is successfully usedto optimize the extraction of Jianpi Yiqi decoctio.
2. The middle level dose of the Qishen Jianpi decoction combined with perindoprilshows the better lowing pressure effect on the caudal artery systolic pressure,after4weeks treatment, it will reach the stable level.
3. The Qishen Jianpi decoction combined with perindopril and the high level dose of Qishen Jianpi decoction can improve SHR cardiac pathology images conformation.By inhibiting the expression of Collagen I and CollagenⅢ so as to reverse theaccumulation of cardiac collagen fibers.
4. By regulating the factors from TGF-β1/Smad signaling pathway and AngII,Qishen Jianpi decoction combined with perindopril could modify the functionobstacle of hearts in the development of arterial blood pressure in SHR.
5. The Qishen Jianpi decoction combined with perindopril and the high level doseof Qishen Jianpi decoction can stimulate the expression of MMP-1, MMP-2and MMP-9to exert its function of lowing blood pressure and protect the hearts.
引文
[1]郭志坤.现代心脏组织学[M].北京:人民卫生出版社,2007:183-200.
[2]邓斌.强心冲剂对扩张型心肌病患者心肌纤维化指标的影响[J].中华中医药学刊，2007,25(6):1149-1150.
[3]杜柏.心衰颗粒对慢性心力衰竭患者超声心动图及心肌纤维化的影响[J].上海中医药大学学报,2006,20(2):21-23.
[4]胡世云,冼绍祥,赵立诚.天麻钩藤饮干预高血压病患者心肌纤维化的临床研究[J].中华实用中西医杂志,2004,4(17):1958.
[5]赵志国.蜈蚣酸性蛋白对血管紧张素-II诱导心肌成纤维细胞增殖和胶原合成的影响[J].中华中医药杂志,2008,23(1):23-26.
[6]赵智明.川芎嗪对新生大鼠心肌成纤维细胞分泌内皮素和一氧化氮的影响[J].内蒙古医学院学报,2006,28(3):202-208.
[7]王健.益气活血法对冠心病患者心肌纤维化影响观察[J].深圳中西医结合杂志,2004,14(2):191.
[8]沈雁.温阳活血化痰法抗心肌纤维化作用及机制研究[J].辽宁中医杂志,2005,32(6):523-524.
[9]李运伦.清热解毒法治疗高血压左室肥厚心肌纤维化43例临床研究[J].山东中医药大学学报,2007,31(6):466-468.
[10] Frey N, Olson EN. Cardiac hypertrophy: the good， the bad， and the ugly[J].AnnuRev Physiol，2003，65(2):45-79.
[11]Levy D, Crarrison RJ， Savage DD, et al. Prognostic implications ofechocardiographically determined left ventricularmass in the Framing-hamHeartstudy [J].N Engl J Med,2010,322:1561-1566.
[12] Berk BC, Fujiwara K, Lehoux S. ECM remodeling in hyperten-sive heartdisease[J].J Clin Invest，2007，117(3):568-575.
[13]张抒扬，戴玉华.全国高血压左室肥厚临床与基础研讨会纪要[J].中华内科杂志，1994，33(2):139-140.
[14]Yamazaki T, Komuro I, Zou Y, et al.Norepinephrine induces the raf-1kinase/mitogen-activated protein kinase cascade through bothα1andβ-adrenoceptors [J].Circulation,2007,95:1260-1268.
[15]Yamazaki T, Komuro I, Zou Y, et al. Protein kinase A and protein ki-nase Csynergistically activate the raf-1kinase/mitogen-activated pro-tein kinasecascade in neonatal rat cardiomyocytes [J].J Mol CellCardiol,2008,29:2491-2501.
[16]蔡鑫.高血压性心肌肥厚的研究进展[J].心血管病学进展.1990,11(4):8-10.
[17]Boon D, Diek JJ, VanmontfransGA.Silent ischaemia and hypertension[J].JHypertens，2000,18:1355-1364.
[18]Hennersdorf MG, Strauer BE.Arterial hypertension and cardiac ar-rhythmias[J].J Hypertens,2001,19:167-177.
[19]Cruickshank J.Reversibility of left ventricular hypertrophy [J].BloodPressure,2002,1(Suppl1):32-34.
[20]Schmieder RE, Messerli FH.Hypertension and the heart [J].J HumHypertens,2000,14:597-604.
[21]Nunez E, Hosoya K, Susic D, et al. Enalapril and losartan reducedcardiac massand improved coronary hemodynamics in SHR [J].Hy-pertension,1997,29(part2):519-524.
[22]Kojima M, Shiojima I, Yamazaki T, et al. AngiotensinⅡreceptor an-tagonistTCV-116induces regression of hypertensive left ventricular hypertrophyin vivoandinhibits the intracellular signaling pathway of stretch-mediated cardiomyocytehypertrophyin vitro[J].Circulation，2004，81:2204-2211.
[23]杨维华,周慎,朱棱.通络熄风利水法对原发性高血压LVR的影响[J].中国中医药科技,2002,9(5):306-307.
[24]范金茹,王行宽,熊国强.高血压LVH中医病名、病位、病因病机探讨[J].中医药通报,2005,4(2):19-21.
[25]喻正科,解发良,朱棱.天丹降压颗粒对高血压病左室肥厚和舒张功能影响的临床研究[J].湖南中医药导报1999,5(12)16-18.
[26]陆曙,吴新欲,翁晓生,等.降防保心片对高血压左室肥厚影响的临床研究[J].南京中医药大学学报,1998,14(1):16-18.
[27]肖梅芳.周端应用活血潜阳法治疗高血压病经验[J].上海中医药杂志,2007,38(4):12-13.
[28]符德玉,罗海明,黄蔚,等.活血潜阳方治疗血瘀、阳亢证高血压病患者左心室肥厚的临床研究[J].上海中医药杂志,2003,37(12):6-9.
[29]王振涛,韩丽华.中医治疗高血压病若干思考[J].中国中西医结合杂志,2003,23(9):709-710.
[30]刘劲松,张榴修,周宜轩.高血压LVH的中医病机学说--LVH的心之体用失常学说[J].中国中医基础医学杂志,2004,10(2):72-75.
[31]Weber K T, Brilla C G, Janicki J S.Myocardial fibrosis:Func-tional significanceand regulatory factors[J].Cardiovasc Res,2009,27:34.
[32]HattoriY, HattoriS, AkimotoK, etal.Globularadiponeetinae–tivatesnuelearfaetor-Bandaetivatlng Protein-1andenhaneesan-giotensinll-indueedproliferat, on, Cardlacfibroblasr,[J].Diabetes,2007,56:804一808.
[33] K-M, tsuyamas, IzumiY.IzumiyaY, etal. Dominant-nega-Tivee-Juninhlbitsrateardiae hypertro Phyindueed byangzotensinUandhypertension[J].eneTher,2006,13:348一355.
[34]BielinokaA, ShivdasaniRA, zhangLQ, etal. Regulation of gene expressionwith double-stranded Phosphorothioateollgonu-eleotides[J].Seienee,1999,250:997一1000
[35]宋国杰.现代心血管病学的新领域———分子心脏病学[M].天津:天津科学技术出版社,2001:117.
[36]WeberKT. Cardiac interstitium in heart and disease:The fibrillar collagennet-work [J]. JAm CollCardio,2009,13(7):1637-1652.
[37]SchwartzAJ, W ilson DA, WeberKT. Factors regulating collagen synthesis anddegradation during second-intention healing ofwounds in the thoracic region andthe distal aspect of the forelimb of horses[J].Am JVetRes,2002,63(11):1564-1570.
[38] Conrad CH， BrooksWW， Hayes JA.Myocardial fibrosis and stiffnesswithhyper-trophy and heart failure in the spontaneously hypertensive rat[J].Circulation,1995,91(1):161-170.
[39] Sugden PH, Clerk A. Cellular mechanismsof cardiac hypertrophy[J]·J Mol Med,1998,76(11):725·
[40]王长谦,谢秀兰,丁弘毅,等·压力负荷增高性心肌肥厚大鼠心肌胶原网络重构及Ⅰ、Ⅲ型胶原mRNA表达的变化[J].心脏杂志,2001,13(5):369·
[41]盛瑞,顾振纶,谢梅林,等.EGCG抑制心肌肥厚胶原生成和细胞增殖[J].中国药理学通报,2006,22(9):1095.
[42]Claridge MW, Hobbs SD, Quick CR, et al. ACE inhibitors increase typeⅢcollagensynthesis:a potential explanation for reduction in acute vascular events by ACEinhibitors.Eur J Vasc Endovasc Surg,2004,28:67-70.
[43]Magga J, Puhakka M, Hietakorpi S, et al. Atrial natriuretic peptide, B-typenatriuretic peptide, and serum collagen markers after acute myocardialinfarction.J Appl Physiol,2004,96:1306-1311.
[44]Sun Y.Myocardial repair/remodelling following infarction: roles of localfactors. Cardiovasc Res,2009,81(3):482—490
[45]Azar ST, Salti I, ZantoutMS, et al.Alteration in plasma transforming growthfactor beta in normoalbuinuric type1and type2diabetic patient[J]. JClinEndocrinolMetab,2002,85(12):4680-1.
[46]MorriseyK, EvansK, Wakefield L, et al.Tanslational regulation of renalproximal tubularepithelial cell transforming growth factor-betalgeneration byinsulin[J].Am J Patho,2007,159(5):1905-15.
[47]SinghR, SongRH, AlaviN, etal.High glucosematrixmetalloproteinase-2activityin ratmesangial cells via transforming growth factor-betal [J].ExpNephro,l2001;9(4):249-57·
[48]刘承云,毛焕元,张银环.血清Ⅰ型、Ⅲ型前胶原端肽浓度在心血管疾病中的意义[M].国外医学心血管分册,1997,24(4):18-20.
[49]Diez J, LaviadesC, MyaorG. Increase serum concentrations of procollagenpeptides in essential hypertention. Relation to cardiac alterations[J].Cardiology,2000,35(2):121-127.
[50]QuerejetaR, VaroN, Lopez B, et al. Serum carboxy-terminal propeptide ofprocollagen type I is a marker ofmyocardial fibrosis in hypertensive heartdisease[J].Circulation,2010,101(14):1729-1735.
[51]陈灏珠.实用内科学[M].第12版.北京:人民卫生出版社，2005:21526-1527.
[52] RaizadaMK, Ferreira AJ. ACE2: a new target for cardiovascular diseasetherapeutics[J].J Cardiovasc Pharmacol,2007,50(2):112-119.
[53] StrothU, UngerT. The renin-angiotensin system and its receptors [J]. JCardiovasc Pharmacol,1999,33(Supp11): S21-S28.
[54]TipnisSR, HooperNM, HydeR, etal. A human homolog ofangiotensin-convertingenzyme-cloning and functional expression as acaptopril-insensitivecarboxypeptidase[J].J Biol Chem,2000,275(43):33238-33243.
[55]曾武涛,马虹,鲁伟,等.血管紧张素-(1-7)在血管紧张素Ⅱ诱导心肌细胞肥大中的作用[J].中华心血管病杂志,2008,28(6):460-463.
[56] Ferreira AJ, Santos RA. Cardiovascular actions ofangiotensin-(1-7)[J].BrazJMed BiolRes,2005,38(4):499-507.
[57]Brilla CG, WeberKT.Reactive and reparativemyocardial fibrosis in arterialhypertention[J].Cardiovasc Res,2002,26(7):671-683.
[58]Pathak M, Sarkar S, Vellaichamy E, et al.Role of myocytes in myocar-dialcollagen production[J].Hypertension,2001,37(3):833-840.
[59]Matsusaka T, Katori H, Inagami T, et al.Communication between myocytes andfibroblasts in cardiac remodeling in angiotensin chimeria mice[J].J Clin Invest,1999,103(10):1451-1458.
[60]Schorb W, Booz GW, Dostal DE, et al.AngiotensinⅡis mitogenic in neonatal ratcardiac fibroblasts[J].Circ Res,1993,72(6):1245-1254.
[61]Brilla CG, Zhou G, Matsubura L, et al.Collagen metabolism in cultured adultrat cardiac fibroblasts:Response to AngiotensinⅡand angosterone[J].J Mol CellCardiol,2009,26(7):809-820.
[62]张萍,何国祥,迟洛湘,等.血管紧张素受体与心肌胶原代谢关系的研究[J].高血压杂志,2001,9(2):148-150.
[63]Varo N, Iraburu MJ, Varela M, et al.Chronic AT1Blockade Stimulates Extracellular CollagenType I Degradation and ReversesMyocardial Fibrosis inSpontaneously Hypertensive Rats[J].Hypertension,2000,35(6):1197-1220.
[64]汤健,魏英杰.心血管活性物质与心血管疾病[M].第1版.北京:北京医科大学协和医科大学联合出版社,1997:180-184.
[65]Kawano H, DoYS, KawanoY, et al.AngiotensinⅡhasmultiple profibrotic effectsin human cardiac fibroblasts[J].Circulation,2000,101(10):1130-1137.
[66]Wei CC, Meng QC, Palmer R, et al.Evidence for Angiotensin-Converting enzymeand Cymase-Mediated AngiotensinⅡformation in the interstitial fluid space of thedog heart in vivo[J].Circulation,1999,99(19):2583-2589.
[67]GrossML, HeissN, WeckbachM, etal.ACE-inhibition is superior to endothelinAreceptorblockade in preventing abnormal capillary supply and fibrosis of the heartin experimental diabetes[J].Diabetologia,2004,47(2):316-324.
[68] Toblli JE, Cao G, DeRosa G, et al. Reduced cardiac expression of plasminogenactivator inhibitor1and transforming growth factor beta-1in obese Zucker ratsby perindopril[J].Heart,2005,91(1):80-86.
[69] Diez J, Querejeta R, Lopez B, et al. Losartan-dependent regression ofmyocardial fibrosis is associated with reduction of left ventricularchamberstiffness in hypertensive patients[J]. Circulation,2002,105(21):2512-2517.
[70] TachlkawaH, KodamaM.HuiL, etal.Angiotensin I type1receptor blocker,valsartan. prevented cardiac fibrosis in rat cardiomyopathy afterautoimmunemyocarditis[J]. JCardiovasc Pharmaco1,2003,41(Suppl1):105-110.
[71] Lim DS,Lutucuta S, Bachireddy P, et al. AngiotensinⅡblockadereversesmyocardial fibrosis in a transgenicmousemodel of human hypertrophiccardiomyopathy[J]. Circulation,2001,103(6):789-791.
[72] Kim S, YoshiyamaM, Lzumi Y, etal. Effects of combination of ACE inhibitor andangiotensin receptor blocker on cardiac remodeling, cardiac function, and survivalin rat heart failure[J]. Circulation,2001,103(1):148-154.
[73] Li Z, Iwai M, Wu L, et al. Role ofAT2receptor in the brain in regulation ofblood pressure andwater intake[J].Am JPhysiol-HeartCirc Physio, l2003,284(1):H116-H121.
[74] Jing-Ji JIN, Jun NAKURA, Zhihong WU, et al. Association of Angiotensin II Type2Receptor Gene Variant with Hypertension. Hypertens Res,2003,26:547-552.
[75] Jugdutt BI, Menon V. Upregulation of angiotensinⅡtype2receptor andlimitation ofmyocardial stunning by angiotensinⅡtype1receptor blockers duringreperfused myocardial infarction in therat[J]. J Cardiovasc PharmacolTher,2003,8(3):217-226.
[76] Brede M, Roell W, Ritter O,et al.Cardiac hypertrophy is associated withdecreased eNOS expression in angiotensin AT2receptor-deficientmice[J].Hypertension,2010,42(6):1177-1182.
[77] Kurisu S, Ozono R, Oshima T, et al. Cardiac angiotensin II type2receptoractivateskin in/NO system and inhihits fibrosis[J].Hypertension,2003,41(1):99-107.
[78] Batenburg WW, Garrelds IM, Bernasconi CC, et al.AngiotensinⅡtype2receptor-mediated vasodilation in human coronary microarteries[J].Circulation,2004,109(19):2296-30113.
[79] Hitoshi S, Kazuhira M, Hiroyuki Y, et al.Altered effects ofan-giotensinⅡtype1and type2receptorblockerson cardiac norep-in-ephrine release and inotropicresponse during cardiac sympa-thetic nerve stimulation in aorto-caval shuntrats[J]. Circ J,2004,68(6):683-693.
[80]Lakó-FutóZ, Szokodi I, Sármán B, et al.Evidence for a functional role ofangiotensinⅡtype2receptor in the cardiac hypertrophic process in vivo in therat heart[J].Hypertension,2003,108(19):2414-2422.
[81] Matsumoto T, Ozono R. Oshima Tetal Type2angiotensinⅡreceptor isdownregulated in cardiomyocytes of patients with heart failure.Cardiovas Res,2002,46:73.
[82]Kingsley DM.The TGF-beta superfamily:new members, new receptors and newgenetictestsof function in different organisms[J].GenesDev,1994,8(2):133-146.
[83]Archer SJ, Bax A, Roberts AB, et al.Transforming growth factor beta1:secondarystructure as determined by heteronuclear magnetic resonancespectroscopy[J].Biochemistry,1993,32(4):1164-1171.
[84]Jennings JC, Mohan S, LinkhartTA, et al.Comparison of the biological actionsofTGFbeta-1andTGFbeta-2:differential activity in endothelial cells[J].J CellPhysiol，2010，137(1):167-172.
[85]Burt DW, Jakowlew SB.Correction:a new interpretation of a chickentransforminggrowth factorβ4complementary DNA[J].Mol Endocrinol,1992,6(6):989-992.
[86] Qian SW, Burmester JK, TsangML, et al.Binding affinity of transforming growthfactor-beta for its type II receptor is determined by the Cterminal region of themolecule[J].J Biol Chem,1996,271(48):30656-30662.
[87] Khalil N.TGF-beta:from latent to active[J].Microbes Infect,1999,1(15):1255-1263.
[88]Yu Q,Stamenkovic I.Cell surface-localized matrixmetalloproteinase-9proteolytically activatesTGF-beta and promotes tumor invasionand an-giogenesis[J].Genes Dev,2009,14(2):163-176.
[89]Crawford SE, StellmachV, Murphy-Ullrich JE, et al.Thrombospondin-1is a majoractivator of TGF-beta1in vivo[J].Cell,1998,93(7):1159-1170.
[90]Miller LA, Barnett NL, Sheppard D, et al.Expression of the beta6in-tegrinsubunit is associated with sites of neutrophil influx in lung epi-thelium[J].JHistochem Cytochem,2001,49(1):41-48.
[91]黄文林,朱孝峰.信号转导[M].北京:人民卫生出版社,2005:203-218.
[92] Sirard C, Kim S, Mirtsos C，et al.Targeted disruption in murine cells revealsvariable requirement for Smad4in transforminggrowth factorβ-relatedsignaling[J].J Biol Chem,2000,275(3):2063-2070.
[93] Nakajima Y, Yamagishi T, Nakamura H, et al.An autocrine function fortransforming growth factor-beta3in the transformation of atrioventricular canalendocardium into mesenchyme during chick heart development[J].Dev Biol,1998,194(1):99-113.
[94] Li JM, Brooks G.Differential portein expression and subcellular distributionof TGF beta1，beta2，beta3in cardiomyocytes during pressures overload inducedhypertrophy[J]. J Mol Cell Cardiol,1997,29(8):2213-2224.
[95] Li G, Li R K, Mickle DA, et al. Elevated insulin-like growth factor-1andtransforming growth factor-beta1and their receptors in patients with idiopathichypertrophic obstructive cardiomyopathy:apossible mechanism[J].Circulation,1998,98(7):Ⅱ-114-Ⅱ-150.
[96] Zhao J, Sime PJ, Bringas P, et al.Adenovirus-mediated decorin gene transferprevents TGF-beta induced inhibition of lung morphogenesis[J].Am J Physiol,1999,277(2):412-422.
[97] Ueno H, Sakamoto T, Nakamure T, et al.A soluble transforming growth factor-betareceptor expressed in muscle prevents liver fibrogenesis and dysfunction inrats[J].Hum Genet Ther,2000,11(1):33-42.
[98]武多娇.麝香保心丸减轻自发性高血压大鼠心肌纤维化的研究[J].中国中西医结合杂志,2005,25(4):350-353.
[99]程志清.通心络对病毒性心肌炎慢性期小鼠心肌纤维化细胞因子TGF、PDGF表达的影响[J].中国中医药科技,2007,14(2):88-89.
[100]赵艳峰.当归对大鼠心肌梗死后心肌纤维化的影响及机制[J].中国病理生理杂志,2005,22(10):1965-1969.
[101]黄力.中药降压脉净对心肌组织转化生长因子-B1的影响及干预心肌纤维化的作用[J].中国心血管病研究,2007,9(5):688-691.
[102]严米娅.阿魏酸钠抑制心肌成纤维细胞增殖的作用及机制[J].武汉大学学报(医学版),2006,27(2):148-151.
[103]刘旺.细胞因子在慢性病毒性心肌炎心肌纤维化作用及清心II号的干预作用[J].浙江中医药大学学报,2008,32(2):165-168.
[104]张海啸.TGF-β信号传导通路与心肌纤维化[J].中日友好医院学报,2007,21(2):110-112.
[105]王爽,张钰泉,张幸,等.HPLC-ELSD法测定黄芪汤颗粒剂中黄芪甲苷的含量[J].长春中医药大学学报,2011,21(6):1048-1049.
[106]赵建邦,马潇.HPLC-ELSD测定补肺丸中黄芪甲苷的含量[J].中国实验方剂学杂志,2010,16(14):89-90.
[107]赵丹琳.蒸发光散射测定黄芪固本胶囊中黄芪甲苷含量[J].湖北中医杂志,2011,33(11):66-67.
[108]张彤,黄顺旺.HPLC-ELSD测定前列舒乐颗粒中的黄芪甲苷[J].中国实验方剂学杂志,2011,17(7):82-83.
[109]何军,奉建芳,庞家忠,等.星点设计-效应面法优化水飞蓟素固体脂质纳米粒的制备[J].中国医药工业杂志,2005,36(1):18-21．
[110]柳俊,张建军.Box-Behnken效应面法优化白芍配方颗粒提取工艺[J].中国实验方剂学杂志,2011,17(1):9-13.
[111]张立明,张霞,郑传莉,等.响应曲面法优化三叶青总黄酮提取工艺研究[J].时珍国医国药,2012,21(10):2588-2590.
[112]田振坤,马英丽,吴伦,等.星点设计-效应面法优化北五味子果实提取工艺[J].中国实验方剂学杂志,2011,17(23):28-31.
[113] Berk BC, Fujiwara K, Lehoux S. ECM remodeling in hyperten-sive heartdisease[J].J Clin Invest,2007,117(3):568-575.
[114]郭志坤.现代心脏组织学[M].北京:人民卫生出版社,2007:183-200.
[115]陈灏珠.实用内科学[M].第12版.北京:人民卫生出版社,2005:21526-1527.
[116] RaizadaMK, Ferreira AJ. ACE2: a new target for cardiovascular diseasetherapeutics[J].J Cardiovasc Pharmacol,2007,50(2):112-119.
[117]曾武涛,马虹,鲁伟,等.血管紧张素-(1-7)在血管紧张素Ⅱ诱导心肌细胞肥大中的作用[J].中华心血管病杂志,2000,28(6):460-463.
[118] Ferreira AJ, Santos RA. Cardiovascular actions ofangiotensin-(1-7)[J].BrazJMed BiolRes,2005,38(4):499-507.
[119] Li G, Li R K, Mickle DA, et al.Elevated insulin-like growth factor-1andtransforming growth factor-beta1and their receptors in patients with idiopathichypertrophic obstructive cardiomyopathy:a possible mechanism[J].Circulation,1998,98(7):Ⅱ-114-Ⅱ-150.
[120]黄文林,朱孝峰.信号转导[M].北京:人民卫生出版社,2005:203-218.
[121]Schneiders D, Heger J, Best P, et a.l SMAD proteins are involved in apoptosisinduction in ventricular cardiomyocytes. Cardiovasc Res,2005,67:87~96.
[122]Ruiz-OrtegaM, Rodríguez-Vita J, Sanchez-Lopez E, et a.l TGF-βsignaling invascular fibrosis. CardiovascRes,2007,74:196~206.
[123] Berk BC, Fujiwara K, Lehoux S. ECM remodeling in hyperten-sive heartdisease[J].J Clin Invest,2007,117(3):568.
[124]胡小勤,曾学文,唐亚平,等.补阳还五汤、天麻钩藤饮与高血压病气虚血瘀证、肝阳上亢证关联性研究[J].中国实验方剂学杂志,2011,17(18):203.
[125]SchwartzAJ, W ilson DA, WeberKT. Factors regulating collagen synthesis anddegradation during second-intention healing ofwounds in the thoracic region andthe distal aspect of the forelimb of horses [J].Am JVetRes,2009,63(11):1564.
[126]盛瑞,顾振纶,谢梅林,等·EGCG抑制心肌肥厚胶原生成和细胞增殖[J].中国药理学通报,2006,22(9):1095·
[127]Claridge MW, Hobbs SD, Quick CR, et al. ACE inhibitors increase typeⅢcollagensynthesis:a potential explanation for reduction in acute vascular events by ACEinhibitors.Eur J Vasc Endovasc Surg,2004,28:67.
[128]Magga J,Puhakka M,Hietakorpi S,et al.Atrial natriuretic peptide,B-typenatriuretic peptide,and serum collagen markers after acute myocardial infarction.JAppl Physiol,2004,96:1306.
[129]B ck M，Ketelhuth D F，Agewall S． Matrix metalloproteinases inatherothrombosis[J].Prog Cardiovasc Dis,2010,52(5):410．
[130]Li X, Wu J F. Recent developments in patent anti-cancer agents targeting thematrix metalloproteinases (MMPs)[J]. Recent Pat Anticancer DrugDiscov,2010,5(2):109.
[131]陈羽.黄芪皂甙Ⅳ对大鼠心肌成纤维细胞胶原影响[J].实用儿科临床杂志,2007,22(7):553.
[132]张召才.黄芪甲甙对慢性心肌炎心肌纤维化的影响[J].中国中西医结合杂志,2007,27(8):728.
[133]邱泽安.天麻钩藤饮治疗社区高血压病[J].中国实验方剂学杂志,2011,17(15):252.
[134]李赛美.加味桃核承气汤及不同提取物对糖尿病大鼠心肌纤维化的影响[J].南京中医药大学学报,2005,21(4):236.
[2]邓斌.强心冲剂对扩张型心肌病患者心肌纤维化指标的影响[J].中华中医药学刊，2007,25(6):1149-1150.
[3]杜柏.心衰颗粒对慢性心力衰竭患者超声心动图及心肌纤维化的影响[J].上海中医药大学学报,2006,20(2):21-23.
[4]胡世云,冼绍祥,赵立诚.天麻钩藤饮干预高血压病患者心肌纤维化的临床研究[J].中华实用中西医杂志,2004,4(17):1958.
[5]赵志国.蜈蚣酸性蛋白对血管紧张素-II诱导心肌成纤维细胞增殖和胶原合成的影响[J].中华中医药杂志,2008,23(1):23-26.
[6]赵智明.川芎嗪对新生大鼠心肌成纤维细胞分泌内皮素和一氧化氮的影响[J].内蒙古医学院学报,2006,28(3):202-208.
[7]王健.益气活血法对冠心病患者心肌纤维化影响观察[J].深圳中西医结合杂志,2004,14(2):191.
[8]沈雁.温阳活血化痰法抗心肌纤维化作用及机制研究[J].辽宁中医杂志,2005,32(6):523-524.
[9]李运伦.清热解毒法治疗高血压左室肥厚心肌纤维化43例临床研究[J].山东中医药大学学报,2007,31(6):466-468.
[10] Frey N, Olson EN. Cardiac hypertrophy: the good， the bad， and the ugly[J].AnnuRev Physiol，2003，65(2):45-79.
[11]Levy D, Crarrison RJ， Savage DD, et al. Prognostic implications ofechocardiographically determined left ventricularmass in the Framing-hamHeartstudy [J].N Engl J Med,2010,322:1561-1566.
[12] Berk BC, Fujiwara K, Lehoux S. ECM remodeling in hyperten-sive heartdisease[J].J Clin Invest，2007，117(3):568-575.
[13]张抒扬，戴玉华.全国高血压左室肥厚临床与基础研讨会纪要[J].中华内科杂志，1994，33(2):139-140.
[14]Yamazaki T, Komuro I, Zou Y, et al.Norepinephrine induces the raf-1kinase/mitogen-activated protein kinase cascade through bothα1andβ-adrenoceptors [J].Circulation,2007,95:1260-1268.
[15]Yamazaki T, Komuro I, Zou Y, et al. Protein kinase A and protein ki-nase Csynergistically activate the raf-1kinase/mitogen-activated pro-tein kinasecascade in neonatal rat cardiomyocytes [J].J Mol CellCardiol,2008,29:2491-2501.
[16]蔡鑫.高血压性心肌肥厚的研究进展[J].心血管病学进展.1990,11(4):8-10.
[17]Boon D, Diek JJ, VanmontfransGA.Silent ischaemia and hypertension[J].JHypertens，2000,18:1355-1364.
[18]Hennersdorf MG, Strauer BE.Arterial hypertension and cardiac ar-rhythmias[J].J Hypertens,2001,19:167-177.
[19]Cruickshank J.Reversibility of left ventricular hypertrophy [J].BloodPressure,2002,1(Suppl1):32-34.
[20]Schmieder RE, Messerli FH.Hypertension and the heart [J].J HumHypertens,2000,14:597-604.
[21]Nunez E, Hosoya K, Susic D, et al. Enalapril and losartan reducedcardiac massand improved coronary hemodynamics in SHR [J].Hy-pertension,1997,29(part2):519-524.
[22]Kojima M, Shiojima I, Yamazaki T, et al. AngiotensinⅡreceptor an-tagonistTCV-116induces regression of hypertensive left ventricular hypertrophyin vivoandinhibits the intracellular signaling pathway of stretch-mediated cardiomyocytehypertrophyin vitro[J].Circulation，2004，81:2204-2211.
[23]杨维华,周慎,朱棱.通络熄风利水法对原发性高血压LVR的影响[J].中国中医药科技,2002,9(5):306-307.
[24]范金茹,王行宽,熊国强.高血压LVH中医病名、病位、病因病机探讨[J].中医药通报,2005,4(2):19-21.
[25]喻正科,解发良,朱棱.天丹降压颗粒对高血压病左室肥厚和舒张功能影响的临床研究[J].湖南中医药导报1999,5(12)16-18.
[26]陆曙,吴新欲,翁晓生,等.降防保心片对高血压左室肥厚影响的临床研究[J].南京中医药大学学报,1998,14(1):16-18.
[27]肖梅芳.周端应用活血潜阳法治疗高血压病经验[J].上海中医药杂志,2007,38(4):12-13.
[28]符德玉,罗海明,黄蔚,等.活血潜阳方治疗血瘀、阳亢证高血压病患者左心室肥厚的临床研究[J].上海中医药杂志,2003,37(12):6-9.
[29]王振涛,韩丽华.中医治疗高血压病若干思考[J].中国中西医结合杂志,2003,23(9):709-710.
[30]刘劲松,张榴修,周宜轩.高血压LVH的中医病机学说--LVH的心之体用失常学说[J].中国中医基础医学杂志,2004,10(2):72-75.
[31]Weber K T, Brilla C G, Janicki J S.Myocardial fibrosis:Func-tional significanceand regulatory factors[J].Cardiovasc Res,2009,27:34.
[32]HattoriY, HattoriS, AkimotoK, etal.Globularadiponeetinae–tivatesnuelearfaetor-Bandaetivatlng Protein-1andenhaneesan-giotensinll-indueedproliferat, on, Cardlacfibroblasr,[J].Diabetes,2007,56:804一808.
[33] K-M, tsuyamas, IzumiY.IzumiyaY, etal. Dominant-nega-Tivee-Juninhlbitsrateardiae hypertro Phyindueed byangzotensinUandhypertension[J].eneTher,2006,13:348一355.
[34]BielinokaA, ShivdasaniRA, zhangLQ, etal. Regulation of gene expressionwith double-stranded Phosphorothioateollgonu-eleotides[J].Seienee,1999,250:997一1000
[35]宋国杰.现代心血管病学的新领域———分子心脏病学[M].天津:天津科学技术出版社,2001:117.
[36]WeberKT. Cardiac interstitium in heart and disease:The fibrillar collagennet-work [J]. JAm CollCardio,2009,13(7):1637-1652.
[37]SchwartzAJ, W ilson DA, WeberKT. Factors regulating collagen synthesis anddegradation during second-intention healing ofwounds in the thoracic region andthe distal aspect of the forelimb of horses[J].Am JVetRes,2002,63(11):1564-1570.
[38] Conrad CH， BrooksWW， Hayes JA.Myocardial fibrosis and stiffnesswithhyper-trophy and heart failure in the spontaneously hypertensive rat[J].Circulation,1995,91(1):161-170.
[39] Sugden PH, Clerk A. Cellular mechanismsof cardiac hypertrophy[J]·J Mol Med,1998,76(11):725·
[40]王长谦,谢秀兰,丁弘毅,等·压力负荷增高性心肌肥厚大鼠心肌胶原网络重构及Ⅰ、Ⅲ型胶原mRNA表达的变化[J].心脏杂志,2001,13(5):369·
[41]盛瑞,顾振纶,谢梅林,等.EGCG抑制心肌肥厚胶原生成和细胞增殖[J].中国药理学通报,2006,22(9):1095.
[42]Claridge MW, Hobbs SD, Quick CR, et al. ACE inhibitors increase typeⅢcollagensynthesis:a potential explanation for reduction in acute vascular events by ACEinhibitors.Eur J Vasc Endovasc Surg,2004,28:67-70.
[43]Magga J, Puhakka M, Hietakorpi S, et al. Atrial natriuretic peptide, B-typenatriuretic peptide, and serum collagen markers after acute myocardialinfarction.J Appl Physiol,2004,96:1306-1311.
[44]Sun Y.Myocardial repair/remodelling following infarction: roles of localfactors. Cardiovasc Res,2009,81(3):482—490
[45]Azar ST, Salti I, ZantoutMS, et al.Alteration in plasma transforming growthfactor beta in normoalbuinuric type1and type2diabetic patient[J]. JClinEndocrinolMetab,2002,85(12):4680-1.
[46]MorriseyK, EvansK, Wakefield L, et al.Tanslational regulation of renalproximal tubularepithelial cell transforming growth factor-betalgeneration byinsulin[J].Am J Patho,2007,159(5):1905-15.
[47]SinghR, SongRH, AlaviN, etal.High glucosematrixmetalloproteinase-2activityin ratmesangial cells via transforming growth factor-betal [J].ExpNephro,l2001;9(4):249-57·
[48]刘承云,毛焕元,张银环.血清Ⅰ型、Ⅲ型前胶原端肽浓度在心血管疾病中的意义[M].国外医学心血管分册,1997,24(4):18-20.
[49]Diez J, LaviadesC, MyaorG. Increase serum concentrations of procollagenpeptides in essential hypertention. Relation to cardiac alterations[J].Cardiology,2000,35(2):121-127.
[50]QuerejetaR, VaroN, Lopez B, et al. Serum carboxy-terminal propeptide ofprocollagen type I is a marker ofmyocardial fibrosis in hypertensive heartdisease[J].Circulation,2010,101(14):1729-1735.
[51]陈灏珠.实用内科学[M].第12版.北京:人民卫生出版社，2005:21526-1527.
[52] RaizadaMK, Ferreira AJ. ACE2: a new target for cardiovascular diseasetherapeutics[J].J Cardiovasc Pharmacol,2007,50(2):112-119.
[53] StrothU, UngerT. The renin-angiotensin system and its receptors [J]. JCardiovasc Pharmacol,1999,33(Supp11): S21-S28.
[54]TipnisSR, HooperNM, HydeR, etal. A human homolog ofangiotensin-convertingenzyme-cloning and functional expression as acaptopril-insensitivecarboxypeptidase[J].J Biol Chem,2000,275(43):33238-33243.
[55]曾武涛,马虹,鲁伟,等.血管紧张素-(1-7)在血管紧张素Ⅱ诱导心肌细胞肥大中的作用[J].中华心血管病杂志,2008,28(6):460-463.
[56] Ferreira AJ, Santos RA. Cardiovascular actions ofangiotensin-(1-7)[J].BrazJMed BiolRes,2005,38(4):499-507.
[57]Brilla CG, WeberKT.Reactive and reparativemyocardial fibrosis in arterialhypertention[J].Cardiovasc Res,2002,26(7):671-683.
[58]Pathak M, Sarkar S, Vellaichamy E, et al.Role of myocytes in myocar-dialcollagen production[J].Hypertension,2001,37(3):833-840.
[59]Matsusaka T, Katori H, Inagami T, et al.Communication between myocytes andfibroblasts in cardiac remodeling in angiotensin chimeria mice[J].J Clin Invest,1999,103(10):1451-1458.
[60]Schorb W, Booz GW, Dostal DE, et al.AngiotensinⅡis mitogenic in neonatal ratcardiac fibroblasts[J].Circ Res,1993,72(6):1245-1254.
[61]Brilla CG, Zhou G, Matsubura L, et al.Collagen metabolism in cultured adultrat cardiac fibroblasts:Response to AngiotensinⅡand angosterone[J].J Mol CellCardiol,2009,26(7):809-820.
[62]张萍,何国祥,迟洛湘,等.血管紧张素受体与心肌胶原代谢关系的研究[J].高血压杂志,2001,9(2):148-150.
[63]Varo N, Iraburu MJ, Varela M, et al.Chronic AT1Blockade Stimulates Extracellular CollagenType I Degradation and ReversesMyocardial Fibrosis inSpontaneously Hypertensive Rats[J].Hypertension,2000,35(6):1197-1220.
[64]汤健,魏英杰.心血管活性物质与心血管疾病[M].第1版.北京:北京医科大学协和医科大学联合出版社,1997:180-184.
[65]Kawano H, DoYS, KawanoY, et al.AngiotensinⅡhasmultiple profibrotic effectsin human cardiac fibroblasts[J].Circulation,2000,101(10):1130-1137.
[66]Wei CC, Meng QC, Palmer R, et al.Evidence for Angiotensin-Converting enzymeand Cymase-Mediated AngiotensinⅡformation in the interstitial fluid space of thedog heart in vivo[J].Circulation,1999,99(19):2583-2589.
[67]GrossML, HeissN, WeckbachM, etal.ACE-inhibition is superior to endothelinAreceptorblockade in preventing abnormal capillary supply and fibrosis of the heartin experimental diabetes[J].Diabetologia,2004,47(2):316-324.
[68] Toblli JE, Cao G, DeRosa G, et al. Reduced cardiac expression of plasminogenactivator inhibitor1and transforming growth factor beta-1in obese Zucker ratsby perindopril[J].Heart,2005,91(1):80-86.
[69] Diez J, Querejeta R, Lopez B, et al. Losartan-dependent regression ofmyocardial fibrosis is associated with reduction of left ventricularchamberstiffness in hypertensive patients[J]. Circulation,2002,105(21):2512-2517.
[70] TachlkawaH, KodamaM.HuiL, etal.Angiotensin I type1receptor blocker,valsartan. prevented cardiac fibrosis in rat cardiomyopathy afterautoimmunemyocarditis[J]. JCardiovasc Pharmaco1,2003,41(Suppl1):105-110.
[71] Lim DS,Lutucuta S, Bachireddy P, et al. AngiotensinⅡblockadereversesmyocardial fibrosis in a transgenicmousemodel of human hypertrophiccardiomyopathy[J]. Circulation,2001,103(6):789-791.
[72] Kim S, YoshiyamaM, Lzumi Y, etal. Effects of combination of ACE inhibitor andangiotensin receptor blocker on cardiac remodeling, cardiac function, and survivalin rat heart failure[J]. Circulation,2001,103(1):148-154.
[73] Li Z, Iwai M, Wu L, et al. Role ofAT2receptor in the brain in regulation ofblood pressure andwater intake[J].Am JPhysiol-HeartCirc Physio, l2003,284(1):H116-H121.
[74] Jing-Ji JIN, Jun NAKURA, Zhihong WU, et al. Association of Angiotensin II Type2Receptor Gene Variant with Hypertension. Hypertens Res,2003,26:547-552.
[75] Jugdutt BI, Menon V. Upregulation of angiotensinⅡtype2receptor andlimitation ofmyocardial stunning by angiotensinⅡtype1receptor blockers duringreperfused myocardial infarction in therat[J]. J Cardiovasc PharmacolTher,2003,8(3):217-226.
[76] Brede M, Roell W, Ritter O,et al.Cardiac hypertrophy is associated withdecreased eNOS expression in angiotensin AT2receptor-deficientmice[J].Hypertension,2010,42(6):1177-1182.
[77] Kurisu S, Ozono R, Oshima T, et al. Cardiac angiotensin II type2receptoractivateskin in/NO system and inhihits fibrosis[J].Hypertension,2003,41(1):99-107.
[78] Batenburg WW, Garrelds IM, Bernasconi CC, et al.AngiotensinⅡtype2receptor-mediated vasodilation in human coronary microarteries[J].Circulation,2004,109(19):2296-30113.
[79] Hitoshi S, Kazuhira M, Hiroyuki Y, et al.Altered effects ofan-giotensinⅡtype1and type2receptorblockerson cardiac norep-in-ephrine release and inotropicresponse during cardiac sympa-thetic nerve stimulation in aorto-caval shuntrats[J]. Circ J,2004,68(6):683-693.
[80]Lakó-FutóZ, Szokodi I, Sármán B, et al.Evidence for a functional role ofangiotensinⅡtype2receptor in the cardiac hypertrophic process in vivo in therat heart[J].Hypertension,2003,108(19):2414-2422.
[81] Matsumoto T, Ozono R. Oshima Tetal Type2angiotensinⅡreceptor isdownregulated in cardiomyocytes of patients with heart failure.Cardiovas Res,2002,46:73.
[82]Kingsley DM.The TGF-beta superfamily:new members, new receptors and newgenetictestsof function in different organisms[J].GenesDev,1994,8(2):133-146.
[83]Archer SJ, Bax A, Roberts AB, et al.Transforming growth factor beta1:secondarystructure as determined by heteronuclear magnetic resonancespectroscopy[J].Biochemistry,1993,32(4):1164-1171.
[84]Jennings JC, Mohan S, LinkhartTA, et al.Comparison of the biological actionsofTGFbeta-1andTGFbeta-2:differential activity in endothelial cells[J].J CellPhysiol，2010，137(1):167-172.
[85]Burt DW, Jakowlew SB.Correction:a new interpretation of a chickentransforminggrowth factorβ4complementary DNA[J].Mol Endocrinol,1992,6(6):989-992.
[86] Qian SW, Burmester JK, TsangML, et al.Binding affinity of transforming growthfactor-beta for its type II receptor is determined by the Cterminal region of themolecule[J].J Biol Chem,1996,271(48):30656-30662.
[87] Khalil N.TGF-beta:from latent to active[J].Microbes Infect,1999,1(15):1255-1263.
[88]Yu Q,Stamenkovic I.Cell surface-localized matrixmetalloproteinase-9proteolytically activatesTGF-beta and promotes tumor invasionand an-giogenesis[J].Genes Dev,2009,14(2):163-176.
[89]Crawford SE, StellmachV, Murphy-Ullrich JE, et al.Thrombospondin-1is a majoractivator of TGF-beta1in vivo[J].Cell,1998,93(7):1159-1170.
[90]Miller LA, Barnett NL, Sheppard D, et al.Expression of the beta6in-tegrinsubunit is associated with sites of neutrophil influx in lung epi-thelium[J].JHistochem Cytochem,2001,49(1):41-48.
[91]黄文林,朱孝峰.信号转导[M].北京:人民卫生出版社,2005:203-218.
[92] Sirard C, Kim S, Mirtsos C，et al.Targeted disruption in murine cells revealsvariable requirement for Smad4in transforminggrowth factorβ-relatedsignaling[J].J Biol Chem,2000,275(3):2063-2070.
[93] Nakajima Y, Yamagishi T, Nakamura H, et al.An autocrine function fortransforming growth factor-beta3in the transformation of atrioventricular canalendocardium into mesenchyme during chick heart development[J].Dev Biol,1998,194(1):99-113.
[94] Li JM, Brooks G.Differential portein expression and subcellular distributionof TGF beta1，beta2，beta3in cardiomyocytes during pressures overload inducedhypertrophy[J]. J Mol Cell Cardiol,1997,29(8):2213-2224.
[95] Li G, Li R K, Mickle DA, et al. Elevated insulin-like growth factor-1andtransforming growth factor-beta1and their receptors in patients with idiopathichypertrophic obstructive cardiomyopathy:apossible mechanism[J].Circulation,1998,98(7):Ⅱ-114-Ⅱ-150.
[96] Zhao J, Sime PJ, Bringas P, et al.Adenovirus-mediated decorin gene transferprevents TGF-beta induced inhibition of lung morphogenesis[J].Am J Physiol,1999,277(2):412-422.
[97] Ueno H, Sakamoto T, Nakamure T, et al.A soluble transforming growth factor-betareceptor expressed in muscle prevents liver fibrogenesis and dysfunction inrats[J].Hum Genet Ther,2000,11(1):33-42.
[98]武多娇.麝香保心丸减轻自发性高血压大鼠心肌纤维化的研究[J].中国中西医结合杂志,2005,25(4):350-353.
[99]程志清.通心络对病毒性心肌炎慢性期小鼠心肌纤维化细胞因子TGF、PDGF表达的影响[J].中国中医药科技,2007,14(2):88-89.
[100]赵艳峰.当归对大鼠心肌梗死后心肌纤维化的影响及机制[J].中国病理生理杂志,2005,22(10):1965-1969.
[101]黄力.中药降压脉净对心肌组织转化生长因子-B1的影响及干预心肌纤维化的作用[J].中国心血管病研究,2007,9(5):688-691.
[102]严米娅.阿魏酸钠抑制心肌成纤维细胞增殖的作用及机制[J].武汉大学学报(医学版),2006,27(2):148-151.
[103]刘旺.细胞因子在慢性病毒性心肌炎心肌纤维化作用及清心II号的干预作用[J].浙江中医药大学学报,2008,32(2):165-168.
[104]张海啸.TGF-β信号传导通路与心肌纤维化[J].中日友好医院学报,2007,21(2):110-112.
[105]王爽,张钰泉,张幸,等.HPLC-ELSD法测定黄芪汤颗粒剂中黄芪甲苷的含量[J].长春中医药大学学报,2011,21(6):1048-1049.
[106]赵建邦,马潇.HPLC-ELSD测定补肺丸中黄芪甲苷的含量[J].中国实验方剂学杂志,2010,16(14):89-90.
[107]赵丹琳.蒸发光散射测定黄芪固本胶囊中黄芪甲苷含量[J].湖北中医杂志,2011,33(11):66-67.
[108]张彤,黄顺旺.HPLC-ELSD测定前列舒乐颗粒中的黄芪甲苷[J].中国实验方剂学杂志,2011,17(7):82-83.
[109]何军,奉建芳,庞家忠,等.星点设计-效应面法优化水飞蓟素固体脂质纳米粒的制备[J].中国医药工业杂志,2005,36(1):18-21．
[110]柳俊,张建军.Box-Behnken效应面法优化白芍配方颗粒提取工艺[J].中国实验方剂学杂志,2011,17(1):9-13.
[111]张立明,张霞,郑传莉,等.响应曲面法优化三叶青总黄酮提取工艺研究[J].时珍国医国药,2012,21(10):2588-2590.
[112]田振坤,马英丽,吴伦,等.星点设计-效应面法优化北五味子果实提取工艺[J].中国实验方剂学杂志,2011,17(23):28-31.
[113] Berk BC, Fujiwara K, Lehoux S. ECM remodeling in hyperten-sive heartdisease[J].J Clin Invest,2007,117(3):568-575.
[114]郭志坤.现代心脏组织学[M].北京:人民卫生出版社,2007:183-200.
[115]陈灏珠.实用内科学[M].第12版.北京:人民卫生出版社,2005:21526-1527.
[116] RaizadaMK, Ferreira AJ. ACE2: a new target for cardiovascular diseasetherapeutics[J].J Cardiovasc Pharmacol,2007,50(2):112-119.
[117]曾武涛,马虹,鲁伟,等.血管紧张素-(1-7)在血管紧张素Ⅱ诱导心肌细胞肥大中的作用[J].中华心血管病杂志,2000,28(6):460-463.
[118] Ferreira AJ, Santos RA. Cardiovascular actions ofangiotensin-(1-7)[J].BrazJMed BiolRes,2005,38(4):499-507.
[119] Li G, Li R K, Mickle DA, et al.Elevated insulin-like growth factor-1andtransforming growth factor-beta1and their receptors in patients with idiopathichypertrophic obstructive cardiomyopathy:a possible mechanism[J].Circulation,1998,98(7):Ⅱ-114-Ⅱ-150.
[120]黄文林,朱孝峰.信号转导[M].北京:人民卫生出版社,2005:203-218.
[121]Schneiders D, Heger J, Best P, et a.l SMAD proteins are involved in apoptosisinduction in ventricular cardiomyocytes. Cardiovasc Res,2005,67:87~96.
[122]Ruiz-OrtegaM, Rodríguez-Vita J, Sanchez-Lopez E, et a.l TGF-βsignaling invascular fibrosis. CardiovascRes,2007,74:196~206.
[123] Berk BC, Fujiwara K, Lehoux S. ECM remodeling in hyperten-sive heartdisease[J].J Clin Invest,2007,117(3):568.
[124]胡小勤,曾学文,唐亚平,等.补阳还五汤、天麻钩藤饮与高血压病气虚血瘀证、肝阳上亢证关联性研究[J].中国实验方剂学杂志,2011,17(18):203.
[125]SchwartzAJ, W ilson DA, WeberKT. Factors regulating collagen synthesis anddegradation during second-intention healing ofwounds in the thoracic region andthe distal aspect of the forelimb of horses [J].Am JVetRes,2009,63(11):1564.
[126]盛瑞,顾振纶,谢梅林,等·EGCG抑制心肌肥厚胶原生成和细胞增殖[J].中国药理学通报,2006,22(9):1095·
[127]Claridge MW, Hobbs SD, Quick CR, et al. ACE inhibitors increase typeⅢcollagensynthesis:a potential explanation for reduction in acute vascular events by ACEinhibitors.Eur J Vasc Endovasc Surg,2004,28:67.
[128]Magga J,Puhakka M,Hietakorpi S,et al.Atrial natriuretic peptide,B-typenatriuretic peptide,and serum collagen markers after acute myocardial infarction.JAppl Physiol,2004,96:1306.
[129]B ck M，Ketelhuth D F，Agewall S． Matrix metalloproteinases inatherothrombosis[J].Prog Cardiovasc Dis,2010,52(5):410．
[130]Li X, Wu J F. Recent developments in patent anti-cancer agents targeting thematrix metalloproteinases (MMPs)[J]. Recent Pat Anticancer DrugDiscov,2010,5(2):109.
[131]陈羽.黄芪皂甙Ⅳ对大鼠心肌成纤维细胞胶原影响[J].实用儿科临床杂志,2007,22(7):553.
[132]张召才.黄芪甲甙对慢性心肌炎心肌纤维化的影响[J].中国中西医结合杂志,2007,27(8):728.
[133]邱泽安.天麻钩藤饮治疗社区高血压病[J].中国实验方剂学杂志,2011,17(15):252.
[134]李赛美.加味桃核承气汤及不同提取物对糖尿病大鼠心肌纤维化的影响[J].南京中医药大学学报,2005,21(4):236.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |