四妙勇安汤促进滋养血管成熟化稳定动脉粥样硬化易损斑块机制研究
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摘要
目的:观察四妙勇安汤对动脉粥样硬化(AS)易损斑块及外膜滋养血管(VV)的干预效应,以VV成熟化为切入点,进一步探讨其起效的机制。方法:采用SPF级健康雄性ApoE-/-小鼠,随机分为模型组、四妙勇安汤组(11.7mg/kg)和辛伐他汀组(2.6mg/kg),均以添加1.1%L-蛋氨酸的高脂饲料喂养8周,C57BL/6小鼠作为正常对照组。药物干预8周后,采用HE染色观察AS斑块情况;通过免疫荧光双标(CD34、Desmin)染色观察主动脉根部斑块内周细胞聚集情况及检测新生滋养血管成熟化密度;通过实时荧光定量PCR技术检测主动脉组织中Dll4、Notch1、Hey1、VEGF mRNA的表达情况。结果:四妙勇安汤可显著减小ApoE-/-小鼠主动脉斑块面积,减少斑块内泡沫细胞浸润,显著降低斑块与管腔面积比,内膜增厚程度明显减轻;可显著升高斑块内CD34、Desmin阳性染色的新生VV密度。四妙勇安汤可上调Dll4、Notch1、Hey1 mRNA的表达,降低VEGF mRNA的表达。结论:四妙勇安汤能减轻小鼠主动脉斑块的病变程度,促进外膜VV周细胞的募集,稳定易损斑块,其机制可能是通过调节Dll4/Notch1/Hey1/VEGF信号通路,同时对VV具有双向调节作用。
Objective: To observe the intervention effect of Simiao Yongan Decoction on atherosclerosis(AS) vulnerable plaque and vasa vasorum(VV), and to explore the mechanism by taking VV maturity as an entry point. Methods: SPF healthy male ApoE-/-mice were randomly divided into model group, Simiao Yongan Decoction group(11.7 mg/kg) and simvastatin group(2.6 mg/kg), all with high-fat diet added 1.1% L-methionine was fed for 8 weeks, and C57 BL/6 mice were used as a control group.After 8 weeks of drug intervention, the plaques of AS were observed by HE staining. The peripheral cells in aortic root plaque were observed by immunofluorescence double staining(CD34, Desmin) and the maturity density of VV was detected. The expression of Dll4, Notch1, Hey1 and VEGF mRNA in aortic tissues was detected by PCR. Results: Simiao Yongan Decoction significantly reduced the area of aortic plaque in ApoE-/-mice, reduced the infiltration of foam cells in plaques, significantly reduced the ratio of plaque to lumen area, and significantly reduced the degree of intimal thickening. Its effect was better than that of simvastatin group;it could significantly increase the density of new VV in CD34 and Desmin positive staining in plaque. Simiao Yongan Decoction could up-regulate the mRNA expression of Dll4, Notch1 and Hey1, and decrease the expression of VEGF, and its effect was better than simvastatin. Conclusion: Simiao Yongan Decoction could alleviate the pathological changes of aortic plaque in mice, promote the recruitment of perivascular VV pericytes, and stabilize vulnerable plaques. The mechanism might be through regulation of Dll4/Notch1/Hey1/VEGF signaling pathway. At the same time, it has a two-way adjustment effect on the VV.
Objective: To observe the intervention effect of Simiao Yongan Decoction on atherosclerosis(AS) vulnerable plaque and vasa vasorum(VV), and to explore the mechanism by taking VV maturity as an entry point. Methods: SPF healthy male ApoE-/-mice were randomly divided into model group, Simiao Yongan Decoction group(11.7 mg/kg) and simvastatin group(2.6 mg/kg), all with high-fat diet added 1.1% L-methionine was fed for 8 weeks, and C57 BL/6 mice were used as a control group.After 8 weeks of drug intervention, the plaques of AS were observed by HE staining. The peripheral cells in aortic root plaque were observed by immunofluorescence double staining(CD34, Desmin) and the maturity density of VV was detected. The expression of Dll4, Notch1, Hey1 and VEGF mRNA in aortic tissues was detected by PCR. Results: Simiao Yongan Decoction significantly reduced the area of aortic plaque in ApoE-/-mice, reduced the infiltration of foam cells in plaques, significantly reduced the ratio of plaque to lumen area, and significantly reduced the degree of intimal thickening. Its effect was better than that of simvastatin group;it could significantly increase the density of new VV in CD34 and Desmin positive staining in plaque. Simiao Yongan Decoction could up-regulate the mRNA expression of Dll4, Notch1 and Hey1, and decrease the expression of VEGF, and its effect was better than simvastatin. Conclusion: Simiao Yongan Decoction could alleviate the pathological changes of aortic plaque in mice, promote the recruitment of perivascular VV pericytes, and stabilize vulnerable plaques. The mechanism might be through regulation of Dll4/Notch1/Hey1/VEGF signaling pathway. At the same time, it has a two-way adjustment effect on the VV.
引文
[1]刘美之,郎艳松,贾振华,等.脉络学说为指导,基于外膜滋养血管探讨动脉粥样硬化发病机制.中国老年病学杂志,2015,35(7):1981-1984
[2]Mulligan-Kehoe M J.The vasa vasorum in diseased and nondiseased arteries.Am J Physiol Heart Circ Physiol,2010,298(2):295-305
[3]李萌,张军平,朱科,等.调控滋养血管新生稳定动脉粥样硬化斑块的中西医治疗进展.世界科学技术-中医药现代化,2017,19(10):1742-1749
[4]漆仲文,张军平,李萌,等.“血-脉-心-神”四位一体理论下动脉粥样硬化性疾病血管稳态维系初探.中华中医药杂志,2018,33(3):863-865
[5]杨萃,袁卓,张军平.四妙勇安汤对血管平滑肌细胞迁移的影响.中华中医药杂志,2013,28(5):1480-1483
[6]张军平,李明,李良军,等.四妙勇安汤调控核因子-κB活性及抑制相关炎症因子的实验研究.中华中医药杂志,2010,25(3):372-376
[7]李萌,张军平,周欢,等.张军平“血-脉-心-神”一体观治疗缺血性心脏病临证思路探析.中华中医药杂志,2018,33(4):1400-1405
[8]徐叔云.药理实验方法学.北京:人民卫生出版社,2002:200-223
[9]Tonar Z,Kural T J,KochováP,et al.Vasa vasorum quantification in human varicose great and small saphenous veins.Ann Anat,2012,94(5):473-481
[10]董震,孙爱军,施海明.动脉粥样硬化斑块滋养血管研究.国际心血管病杂志,2014,41(4):233-235
[11]Eriksson E E.Intravital microscopy on atherosclerosis in apolipoprotein e-deficient mice establishes microvessels as major entry pathways for leukocytes to advanced lesions.Circulation,2011,124(19):2129
[12]Ho-Tin-No?B,Le D J,Gomez D,et al.Early atheroma-derived agonists of peroxisome proliferator-activated receptor-?3trigger intramedial angiogenesis in a smooth muscle cell-dependent manner.Circulation Research,2011,109(9):1003-1014
[13]林志荣,张朝然.血管周细胞与内皮细胞相互关系的研究进展.中国眼耳鼻喉科杂志,2008,18(4):260-261
[14]Amulik A,Abramsson A,Betsholtz C.Endothelial/Pericyte Interactions.Cir Res,2005,97(6):260-261
[15]漆仲文,李萌,张军平.从滋养血管成熟化探讨稳定动脉粥样硬化易损斑块的作用.中国动脉硬化杂志,2017,25(7):737-740
[16]衣慧,王宗仁.Angiopoietin/Tie2系统在血管新生与成熟过程中的表达及意义.心脏杂志,2012,24(4):535-538
[17]Hilbert T,Klaschik S.The angiopoietin/TIE receptor system:Focusing its role for ischemia-reperfusion injury.Cytokine Growth Factor Rev,2015,26(3):281-291
[18]Chen J X,Tuo Q,Liao D F,et al.Inhibition of protein typrosine phosphatase improves angiogenesis via enhancing Ang-2/Tie-2signaling in diabetes.Exp Diabetes Res,2012,20(12):1-10
[19]Ehebauer M,Hayward P,Arias A M.Notch,a universal arbiter of cell fate decisions.Science,2006,314(4):1414-1415
[20]Yan M,Plowman G D.Delta-like 4/Notch signaling and its therapeutic implications.Clin Cancer Res,2007,13(24):7243-7246
[2]Mulligan-Kehoe M J.The vasa vasorum in diseased and nondiseased arteries.Am J Physiol Heart Circ Physiol,2010,298(2):295-305
[3]李萌,张军平,朱科,等.调控滋养血管新生稳定动脉粥样硬化斑块的中西医治疗进展.世界科学技术-中医药现代化,2017,19(10):1742-1749
[4]漆仲文,张军平,李萌,等.“血-脉-心-神”四位一体理论下动脉粥样硬化性疾病血管稳态维系初探.中华中医药杂志,2018,33(3):863-865
[5]杨萃,袁卓,张军平.四妙勇安汤对血管平滑肌细胞迁移的影响.中华中医药杂志,2013,28(5):1480-1483
[6]张军平,李明,李良军,等.四妙勇安汤调控核因子-κB活性及抑制相关炎症因子的实验研究.中华中医药杂志,2010,25(3):372-376
[7]李萌,张军平,周欢,等.张军平“血-脉-心-神”一体观治疗缺血性心脏病临证思路探析.中华中医药杂志,2018,33(4):1400-1405
[8]徐叔云.药理实验方法学.北京:人民卫生出版社,2002:200-223
[9]Tonar Z,Kural T J,KochováP,et al.Vasa vasorum quantification in human varicose great and small saphenous veins.Ann Anat,2012,94(5):473-481
[10]董震,孙爱军,施海明.动脉粥样硬化斑块滋养血管研究.国际心血管病杂志,2014,41(4):233-235
[11]Eriksson E E.Intravital microscopy on atherosclerosis in apolipoprotein e-deficient mice establishes microvessels as major entry pathways for leukocytes to advanced lesions.Circulation,2011,124(19):2129
[12]Ho-Tin-No?B,Le D J,Gomez D,et al.Early atheroma-derived agonists of peroxisome proliferator-activated receptor-?3trigger intramedial angiogenesis in a smooth muscle cell-dependent manner.Circulation Research,2011,109(9):1003-1014
[13]林志荣,张朝然.血管周细胞与内皮细胞相互关系的研究进展.中国眼耳鼻喉科杂志,2008,18(4):260-261
[14]Amulik A,Abramsson A,Betsholtz C.Endothelial/Pericyte Interactions.Cir Res,2005,97(6):260-261
[15]漆仲文,李萌,张军平.从滋养血管成熟化探讨稳定动脉粥样硬化易损斑块的作用.中国动脉硬化杂志,2017,25(7):737-740
[16]衣慧,王宗仁.Angiopoietin/Tie2系统在血管新生与成熟过程中的表达及意义.心脏杂志,2012,24(4):535-538
[17]Hilbert T,Klaschik S.The angiopoietin/TIE receptor system:Focusing its role for ischemia-reperfusion injury.Cytokine Growth Factor Rev,2015,26(3):281-291
[18]Chen J X,Tuo Q,Liao D F,et al.Inhibition of protein typrosine phosphatase improves angiogenesis via enhancing Ang-2/Tie-2signaling in diabetes.Exp Diabetes Res,2012,20(12):1-10
[19]Ehebauer M,Hayward P,Arias A M.Notch,a universal arbiter of cell fate decisions.Science,2006,314(4):1414-1415
[20]Yan M,Plowman G D.Delta-like 4/Notch signaling and its therapeutic implications.Clin Cancer Res,2007,13(24):7243-7246