中药肝复康对实验性肝纤维化大鼠细胞外基质及其相关因子的影响
|
摘要
目的:观察肝复康作用实验性肝纤维化大鼠后血清细胞外基质标志物(Ⅳ型胶原、Ⅲ型前胶原、透明质酸、层粘连蛋白)的变化情况及与细胞外基质降解与沉积有关的基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶抑制因子-2(TIMP-2)基因表达变化。以期深入探讨肝纤维化的发生、发展及逆转过程机制,最终为临床诊断及改善肝纤维化的治疗现状提供新思路。
方法:健康SD大鼠77只,将其随机分为正常对照组(C)10只,模型组(M)11只,模型对照组(MC)11只,低剂量治疗组(LT)10只、中剂量治疗组(MT)12只、高剂量治疗组(HT)12只,预防组(P)11只。除正常对照组外,余鼠造模。皮下注射以橄榄油稀释的10%的四氯化碳,5ml/kg,每周两次,共12周,正常对照组以同样方法皮下注射生理盐水。其中预防组于造模同时以312.5 mg /kg﹒d剂量的肝复康煎剂(主要成分为丹参、黄芪、赤芍、白芍、柴胡等)灌胃12周。高剂量治疗组、中剂量治疗组、低剂量治疗组则于造模第9周后开始分别以3125mg/kg﹒d、312.5mg/kg﹒d、31.25mg/kg﹒d剂量的肝复康灌胃12周。以上各剂量药物均溶于10 ml/kg的生理盐水中。正常对照组及模型对照组则以等容量的生理盐水灌胃。第12周末将模型组、预防组大鼠隔夜禁食,次日称量体重后,以乙醚麻醉腹主动脉采血检测各项指标,处死大鼠,迅速取出肝组织,从肝右叶的中部切取肝组织于-70℃冰箱保存用于RT-PCR。第20周末将其余各组大鼠以同样方法处死,取出肝组织,备用。应用放射免疫分析法检测大鼠血清样本中各项指标的变化情况(Ⅳ型胶原、Ⅲ型前胶原、透明质酸、层粘连蛋白);采用RT-PCR法检测各组大鼠肝组织中基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶抑制因子-2(TIMP-2)基因的表达情况,用Olympus显微摄像系统采集图片,用图像分析软件测定其积分光密度(IOD),以IOD值表示表达水平,对IOD值进行分析和比较。
结果:1、模型组及模型对照组血清Ⅳ型胶原、Ⅲ型前胶原、透明质酸、层粘连蛋白含量较之正常对照组均有明显增高(P=0.000),预防组与正常对照组血清四项指标比较无显著差异(P>0.05)。经不同剂量的中药肝复康干预后,该四项指标血清值均有明显降低(P<0.05),并以中剂量组作用效果最明显;2、基于RT-PCR和其IOD值检测得到的结果提示:MMP-2和TIMP-2在正常对照组表达微弱,在肝脏纤维化模型及模型对照组表达较正常对照组显著增强(P<0.05),经不同剂量的中药肝复康治疗后表达均有明显降低(P<0.05),其中中剂量治疗组降低水平最为显著,其表达水平基本恢复至正常大鼠相应表达水平,治疗效果优于高剂量、低剂量。
结论:①中药肝复康具有较好的预防和治疗肝纤维化作用,并且以中剂量作用效果最为明显,是较为合理的用药剂量。②中药肝复康可以通过抑制胶原合成,减少ECM生成减轻大鼠肝纤维化程度。③中药肝复康可能通过解除MMP-2的抑制因素而提高其活性,以促进胶原降解。④中药肝复康通过调控基因表达水平,抑制MMP-2和TIMP-2mRNA表达,使MMP-2/TIMP-2趋于平衡,从而发挥其抗纤维化的作用。
Objective To observe the changes of serum HA,LN,CIV,PCⅢand MMP-2、TIMP-2 which are correlated with the extracellular matrix (ECM) degradation after the effect of GFK on experimental rats with hepatic fibrosis. To elucidate the expression patterns of MMP-2, TIMP-2. To determin the potential genetic mechanism of hepatic fibrosis’s happen, development and reverse. To provide an adjuvant therapeutic clue for better treatment of human hepatic fibrosis.
Methods Healthy SD rats were divided into seven groups at random including normal control group(n=10),Ganfukang high dose group(n=12),Ganfukang medium dose group (n=12),Ganfukang low dose group(n=10),model group(n=11), model control group(n=11), and preventive group (n=11). All groups except normal control group were subcutaneous injected 10% CCl4(5ml/kg) for 12 weeks and two times each week to obtain hepatic fibrosis and were treated with sodium chloride,GFK3125mg/kg ,GFK312.5mg/kg, GFK31.25mg/kg respectively after nine weeks of CCl4 injection. Model control group were treated with sodium chloride. Preventive group were treated with GFK 312.5mg/kg.d for two week as soon as the model is established. At the end of the twelfth week, rats in model and preventive groups were put to death, reserve the serum to detect four ECM targets. Also fetch out the liver as quickly as possible and reserve in the -70℃refrigeratory for the RT-PCR. The mRNA expression of MMP-2 and TIMP-2 were measured by semi-quantitation reverse transcriptase polymerase chain reaction (RT-PCR). The RT-PCR results was input into the image analysis software to detect the integrated option density(IOD)to show expression level and so to analysis and compare. At the end of the twenties week, the left rats were treated with the same methods as the above. RIA technique were applied to detect the serum levels of CIV,PCⅢ,HA,LN.
Results 1) The serum level of HA,LN,CIV,PCⅢwere higher in the model and model control groups, which has significant difference compare with normal control group (P<0·05). There is no significant difference between preventive and normal control group (P>0·05). The collagen fibrosis decreased significantly in GFK group, this groups has significant difference compare to model group (P<0·05). And the effect of medium dose GFK is the most effective. 2) RT-PCR demonstrated that expression of MMP-2 and TIMP-2 in model group increased significantly compared with normal control group,While the expression of MMP-2 and TIMP-2mRNA in the therapeutic groups reduced significantly compared with model group. The same result we can get is that the effect of medium dose GFK is the most effective.
Conclusion 1) GFK has preferable preventive and therapy effects on rats with hepatic fibrosis. And the effect of medium dose GFK is the most effective. It’s the reasonable dosage.2) GFK can significantly relieve the degree of hepatic fibrosis. This may be through the way that GFK inhibit synthesis of collagen and reduce generation of ECM. 3) GFK can probably remove the MMP-2 inhabitor so to improve the bioactivity of MMP-2 to promote the ECM degradation. 4) GFK may play its anti-fibrosis function through regulates gene expression, repress the expression of MMP-2 and TIMP-2 mRNA to make MMP-2 and TIMP-2 tend to be balanced.
方法:健康SD大鼠77只,将其随机分为正常对照组(C)10只,模型组(M)11只,模型对照组(MC)11只,低剂量治疗组(LT)10只、中剂量治疗组(MT)12只、高剂量治疗组(HT)12只,预防组(P)11只。除正常对照组外,余鼠造模。皮下注射以橄榄油稀释的10%的四氯化碳,5ml/kg,每周两次,共12周,正常对照组以同样方法皮下注射生理盐水。其中预防组于造模同时以312.5 mg /kg﹒d剂量的肝复康煎剂(主要成分为丹参、黄芪、赤芍、白芍、柴胡等)灌胃12周。高剂量治疗组、中剂量治疗组、低剂量治疗组则于造模第9周后开始分别以3125mg/kg﹒d、312.5mg/kg﹒d、31.25mg/kg﹒d剂量的肝复康灌胃12周。以上各剂量药物均溶于10 ml/kg的生理盐水中。正常对照组及模型对照组则以等容量的生理盐水灌胃。第12周末将模型组、预防组大鼠隔夜禁食,次日称量体重后,以乙醚麻醉腹主动脉采血检测各项指标,处死大鼠,迅速取出肝组织,从肝右叶的中部切取肝组织于-70℃冰箱保存用于RT-PCR。第20周末将其余各组大鼠以同样方法处死,取出肝组织,备用。应用放射免疫分析法检测大鼠血清样本中各项指标的变化情况(Ⅳ型胶原、Ⅲ型前胶原、透明质酸、层粘连蛋白);采用RT-PCR法检测各组大鼠肝组织中基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶抑制因子-2(TIMP-2)基因的表达情况,用Olympus显微摄像系统采集图片,用图像分析软件测定其积分光密度(IOD),以IOD值表示表达水平,对IOD值进行分析和比较。
结果:1、模型组及模型对照组血清Ⅳ型胶原、Ⅲ型前胶原、透明质酸、层粘连蛋白含量较之正常对照组均有明显增高(P=0.000),预防组与正常对照组血清四项指标比较无显著差异(P>0.05)。经不同剂量的中药肝复康干预后,该四项指标血清值均有明显降低(P<0.05),并以中剂量组作用效果最明显;2、基于RT-PCR和其IOD值检测得到的结果提示:MMP-2和TIMP-2在正常对照组表达微弱,在肝脏纤维化模型及模型对照组表达较正常对照组显著增强(P<0.05),经不同剂量的中药肝复康治疗后表达均有明显降低(P<0.05),其中中剂量治疗组降低水平最为显著,其表达水平基本恢复至正常大鼠相应表达水平,治疗效果优于高剂量、低剂量。
结论:①中药肝复康具有较好的预防和治疗肝纤维化作用,并且以中剂量作用效果最为明显,是较为合理的用药剂量。②中药肝复康可以通过抑制胶原合成,减少ECM生成减轻大鼠肝纤维化程度。③中药肝复康可能通过解除MMP-2的抑制因素而提高其活性,以促进胶原降解。④中药肝复康通过调控基因表达水平,抑制MMP-2和TIMP-2mRNA表达,使MMP-2/TIMP-2趋于平衡,从而发挥其抗纤维化的作用。
Objective To observe the changes of serum HA,LN,CIV,PCⅢand MMP-2、TIMP-2 which are correlated with the extracellular matrix (ECM) degradation after the effect of GFK on experimental rats with hepatic fibrosis. To elucidate the expression patterns of MMP-2, TIMP-2. To determin the potential genetic mechanism of hepatic fibrosis’s happen, development and reverse. To provide an adjuvant therapeutic clue for better treatment of human hepatic fibrosis.
Methods Healthy SD rats were divided into seven groups at random including normal control group(n=10),Ganfukang high dose group(n=12),Ganfukang medium dose group (n=12),Ganfukang low dose group(n=10),model group(n=11), model control group(n=11), and preventive group (n=11). All groups except normal control group were subcutaneous injected 10% CCl4(5ml/kg) for 12 weeks and two times each week to obtain hepatic fibrosis and were treated with sodium chloride,GFK3125mg/kg ,GFK312.5mg/kg, GFK31.25mg/kg respectively after nine weeks of CCl4 injection. Model control group were treated with sodium chloride. Preventive group were treated with GFK 312.5mg/kg.d for two week as soon as the model is established. At the end of the twelfth week, rats in model and preventive groups were put to death, reserve the serum to detect four ECM targets. Also fetch out the liver as quickly as possible and reserve in the -70℃refrigeratory for the RT-PCR. The mRNA expression of MMP-2 and TIMP-2 were measured by semi-quantitation reverse transcriptase polymerase chain reaction (RT-PCR). The RT-PCR results was input into the image analysis software to detect the integrated option density(IOD)to show expression level and so to analysis and compare. At the end of the twenties week, the left rats were treated with the same methods as the above. RIA technique were applied to detect the serum levels of CIV,PCⅢ,HA,LN.
Results 1) The serum level of HA,LN,CIV,PCⅢwere higher in the model and model control groups, which has significant difference compare with normal control group (P<0·05). There is no significant difference between preventive and normal control group (P>0·05). The collagen fibrosis decreased significantly in GFK group, this groups has significant difference compare to model group (P<0·05). And the effect of medium dose GFK is the most effective. 2) RT-PCR demonstrated that expression of MMP-2 and TIMP-2 in model group increased significantly compared with normal control group,While the expression of MMP-2 and TIMP-2mRNA in the therapeutic groups reduced significantly compared with model group. The same result we can get is that the effect of medium dose GFK is the most effective.
Conclusion 1) GFK has preferable preventive and therapy effects on rats with hepatic fibrosis. And the effect of medium dose GFK is the most effective. It’s the reasonable dosage.2) GFK can significantly relieve the degree of hepatic fibrosis. This may be through the way that GFK inhibit synthesis of collagen and reduce generation of ECM. 3) GFK can probably remove the MMP-2 inhabitor so to improve the bioactivity of MMP-2 to promote the ECM degradation. 4) GFK may play its anti-fibrosis function through regulates gene expression, repress the expression of MMP-2 and TIMP-2 mRNA to make MMP-2 and TIMP-2 tend to be balanced.
引文
1. Bataller R,Brenner DA. Liver fibrosis. J Clin Invest,2005,115:209-218
2. Friedman SL.Liver fibrosis-from bench to bedside. J Hepatol 2003,38 supp l1:S38-53.
3. Gabele E , Brenner DA , Rippe RA.Liver fibrosis : signals leading to the amplification of the fibrogenic hepatic stellate cell. Front Biosci 2003,8:d69-77.
4. Das SK, Vasudevan DM. Genesis of hepatic fibrosis and its biochemical markers. Scand J Clin Lab Invest. 2008; 68(4):260-9. Review.
5. Itagaki T, Shimizu I, Cheng X, et al. Opposing effects of oes-tradiol and progesterone on intracellular pathways and activation processes in the oxidative stress induced activation of culturedrat hepatic stellate cells. Gut,2005, 54: 1782-1789.
6. Lamireau T,Desmouliere A,Bioulac-Sage P,et al.Mechanisms of hepatic fibrogenesis [J].Arch Pediatr,2002,9(4):392-405.
7.胡义,王庆林。国际病理科学与临床杂志。2007,27(1):48-52
8. Van Eyken P,Sciot R,Desmet VJ. Expression of the novel extracellular matrix component tenascin in normal and diseased human liver An immunohisto- chemical study. J Hepatol,2000,11(1):42-52.
9.冯新利,徐贵平,欧阳艳。肝纤维化血清学诊断方法的研究进展。放射免疫学杂志,2003,16(2):111-113
10.何云,王建宾,王宇明。肝纤维化程度诊断方法评价[J]。成都军区医院学报,2002,4(3):35
11.董永杰,彭彦辉,赵晓云,冯运章,孟会彦,李辉。肝纤维化血清学诊断方法的研究进展。医学综述,2007,13(22):1743-1745
12.蔡卫民,陶君,翁红雷,等。血清纤维化指标的影响因素分析[J]。中华肝脏病杂志。2003,11(1):34
13.侯振江,张宗英,周秀梅。蛋白标记物在肝纤维化研究中的应用。中华肝脏病杂志,1998;14(4):200-203
14. Timpl R, Rohde H, Robey PG, Rennard SI, Foidart JM, Martin GR. Laminin-a glycoprotein from basement membranes. J Biol Chem. 1979 Oct 10;254 (19): 9933-7.
15.李春辉,潘理会。肝纤维化发生机制的研究进展。承德医学院学报, 2005,22(1):62-65
16. Chun-Hui Li Dong-Ming Piao Wen-Xie. Morphological and serum hyaluronic acid, laminin and type IV collagen changes in dimethylnitrosamine-induced hepatic fibrosis of rats.世界胃肠病学杂志:英文版。2005,11(48):85-92。
17. Heitor Rosa, Edison Roberto Parise. Is there a place for serum laminin determination in patients with liver disease and cancer?.世界胃肠病学杂志:英文版,2008,14(23)。
18. Parsons CJ, Takashima M, Rippe RA. Molecular mechanisms of hepatic fibrogenesis. J Gastroenterol Hepatol. 2007 Jun; 22 Suppl 1:S79-84. Review.
19. Gressner OA, Weiskirchen R, Gressner AM.Biomarkers of hepatic fibrosis, fibrogenesis and genetic pre-disposition pending between fiction and reality. J Cell Mol Med. 2007 Sep-Oct;11(5):1031-51. Review.
20. PAN Qin, LI Ding—guo, LU Han—ruing, ZHANG Wen—zhu, XU Qin—Fang. Negative regulatory role of somatostatin and octreotide in extracellular matrixes metabolism of rat hepatic stellate cell. Journal of Shanghai Second Medical University, 2008, 20 (2):71-76.
21. Benyon RC,Arthur MJ,.Extracellular matrix degradation and the role of hepatic stellate cells.Semin Liver Dis,2001,21:373-384.
22.徐斌,张振华,黄祝清。血清基质金属蛋白酶及其组织抑制因子与肝纤维化.实用肝脏病杂志,2004,7(1):56-58
23.高润平,齐晓艳。肝纤维化的发生机制与治疗进展。世界华人消化杂志,2006,14(23):2263-2269
24. NIE QH,CHENG YQ,XIE YM. The inhibiting effects of antisense oligonucleotides phosphoroth ioate on gene expression of TMPs in rat liver fibrosis. JG astroentero.2001,7:363.
25.刘海林,王磊。肝纤维化的研究进展。国际消化病杂志。2007,27(4):235-236
1. HammelP,CouvelardA,0’TooleD,RatouisA,SauvanetA,FlejouJF,DegottC,BeighitiJ,BernadesP.VallaD,RusZniewski P,LevyP. Regression of liver fibrosis after biliary drainage in Patients with chronic Pancreatitis and stenosis of the common bile duct. N Engl J Med 2001;344:418-423.
2. Benyon RC, Iredale JP. Is liver fibrosis reversible? Gut 2000; 46: 443-446.
3. Benyon RC, Arthur MJP. Mechanisms of hepatic fibrosis. J Pediatr Gastroente rol Nutr 1998; 27:75-85.
4.姜慧卿,张晓岚。肝纤维化的发生机制。世界华人消化杂志2000;8:687-689
5.张莉娟,王小众,黄月红,陈治新。降钙素基因相关肽、血管紧张素Ⅱ和内皮素在大鼠纤维化中的作用。世界华人消化杂志2001;9:457-459
6. Hayasaka A, Saisho H. Serum markers as tools to monitor liver fibrosis. [J].Digestion, 1998, 59∶381-384.
7.贾继东,尹珊珊。肝纤维化的诊断和治疗进展[J].临床内科杂志,2002,19:244-246
8.张萍。血清LN、CIV、PCⅢ、HA对肝纤维化及肝硬化的诊断价值[J]。中国疗养医学,2002,11(1):50
9.周光德,赵景民。细胞外基质在肝内的代谢与肝纤维化的形成。世界华人消化杂志2002;10(1):57-59
10.蔡卫民,张彬彬,翁红雷等。八项肝纤维化血清标志物比较研究。[J].中华肝脏病杂志2004,12(4)∶219-222.
11.罗瑞红,杨绍基,谢俊强等。肝纤维化血清五项标志物的诊断意义[J].中华肝脏病杂志2001,9(3):148-150
12. Benyon RC, Hovell CJ, Gaca MD, Jones EH, Iredale JP, Arthur MJP. Progelatinas e A is produced and activated by rat hepatic stellate cells and promotes their proliferation. Hepatology 1999;30: 977-986.
13. Nagase H,Woessner JF Jr . Matrix metalloProteinases. J Biol Chem. 1999二30: 21491-21494.
14. MASSOVA IKOTRA LP, FRIDMAN R, MOBASHERY S Matrix metallopro- teinases: structure, evolution, and diversification[ J] FASEB,1998, 12: 1075.
15.朱跃科,王宝恩,申凤俊等。实验性肝纤维化基质金属蛋白酶-2基因表达与酶活性的变化[J]。中华肝脏病杂志,2005,13(7):509-512
16. Pre′aux AM, Mallat A, van Nhieu JT, D′Ortho MP, Hembry RM, Mavier P. Matrix metalloproteinase-2 activation in human hepatic fibrosis regulation by cell-matrix interactions. Hepatology 1999; 30: 944-950.
17. Knittel T, Mehde M, Kobold D, Saile B, Dinter C, Ramadori G. Expressio n patterns of matrix metalloproteinases and their inhibitors in parenchymal and non-parenchymal cells of rat liver: regulation by TNF-αand TGF-β1. J Hepatol 1999; 30:48-60.
18.张亚飞,聂和青,谢玉梅。抑制金属蛋白酶组织抑制因子-2在肝组织中的表达对大鼠肝纤维化的影响[J]。中华肝脏病杂志2006,14:138-140
19. NINOMIYA T, YOON S, NAGANO H,et al. Significance of serum matrix metalloproteinases and their inhibitors on the antifibrogenetic effect of interferorr alfa in chronic hepatitis C patients[J]. Intervirology,2001,44(4): 227-231.
20. GIANNELLI G , BERGAMINI C , MARINOSCI F , et al. Clinical role of MMP-2/TIMP-2 imbalance in hepatocellular carcinoma[J].Int J Cancer,2002, 97(4):425-431.
21. BENYON RC,ARTHUR MJ. Extracellular of matrix degradation and the role of hepatic stellate cells[J].Semin Liver Dis,200l,2l:373-384.
22. Cassiman D, Libbrecht L, Desmet V, Aertsen P, Denef C, Roskams T. Hepatic stellate cell/myofibroblast subpopulations in fibrotic human and rat livers. J Hepatol 2001; 34(suppl,1):20.
23.刘文滨,王吉耀。NF-κB与肝星状细胞凋亡。世界华人消化杂志2001;9:1054-1055
24. ONOM,MLYAMURAM,KYOTANI S,et al. Effect of Shosaiko to ex-tracter on HGF and TGF-beta levels of intraorgans in liver injured rats after partial hepatectomy[J] J Pharm Pharmacol,2000, 52(1): 111.
2. Friedman SL.Liver fibrosis-from bench to bedside. J Hepatol 2003,38 supp l1:S38-53.
3. Gabele E , Brenner DA , Rippe RA.Liver fibrosis : signals leading to the amplification of the fibrogenic hepatic stellate cell. Front Biosci 2003,8:d69-77.
4. Das SK, Vasudevan DM. Genesis of hepatic fibrosis and its biochemical markers. Scand J Clin Lab Invest. 2008; 68(4):260-9. Review.
5. Itagaki T, Shimizu I, Cheng X, et al. Opposing effects of oes-tradiol and progesterone on intracellular pathways and activation processes in the oxidative stress induced activation of culturedrat hepatic stellate cells. Gut,2005, 54: 1782-1789.
6. Lamireau T,Desmouliere A,Bioulac-Sage P,et al.Mechanisms of hepatic fibrogenesis [J].Arch Pediatr,2002,9(4):392-405.
7.胡义,王庆林。国际病理科学与临床杂志。2007,27(1):48-52
8. Van Eyken P,Sciot R,Desmet VJ. Expression of the novel extracellular matrix component tenascin in normal and diseased human liver An immunohisto- chemical study. J Hepatol,2000,11(1):42-52.
9.冯新利,徐贵平,欧阳艳。肝纤维化血清学诊断方法的研究进展。放射免疫学杂志,2003,16(2):111-113
10.何云,王建宾,王宇明。肝纤维化程度诊断方法评价[J]。成都军区医院学报,2002,4(3):35
11.董永杰,彭彦辉,赵晓云,冯运章,孟会彦,李辉。肝纤维化血清学诊断方法的研究进展。医学综述,2007,13(22):1743-1745
12.蔡卫民,陶君,翁红雷,等。血清纤维化指标的影响因素分析[J]。中华肝脏病杂志。2003,11(1):34
13.侯振江,张宗英,周秀梅。蛋白标记物在肝纤维化研究中的应用。中华肝脏病杂志,1998;14(4):200-203
14. Timpl R, Rohde H, Robey PG, Rennard SI, Foidart JM, Martin GR. Laminin-a glycoprotein from basement membranes. J Biol Chem. 1979 Oct 10;254 (19): 9933-7.
15.李春辉,潘理会。肝纤维化发生机制的研究进展。承德医学院学报, 2005,22(1):62-65
16. Chun-Hui Li Dong-Ming Piao Wen-Xie. Morphological and serum hyaluronic acid, laminin and type IV collagen changes in dimethylnitrosamine-induced hepatic fibrosis of rats.世界胃肠病学杂志:英文版。2005,11(48):85-92。
17. Heitor Rosa, Edison Roberto Parise. Is there a place for serum laminin determination in patients with liver disease and cancer?.世界胃肠病学杂志:英文版,2008,14(23)。
18. Parsons CJ, Takashima M, Rippe RA. Molecular mechanisms of hepatic fibrogenesis. J Gastroenterol Hepatol. 2007 Jun; 22 Suppl 1:S79-84. Review.
19. Gressner OA, Weiskirchen R, Gressner AM.Biomarkers of hepatic fibrosis, fibrogenesis and genetic pre-disposition pending between fiction and reality. J Cell Mol Med. 2007 Sep-Oct;11(5):1031-51. Review.
20. PAN Qin, LI Ding—guo, LU Han—ruing, ZHANG Wen—zhu, XU Qin—Fang. Negative regulatory role of somatostatin and octreotide in extracellular matrixes metabolism of rat hepatic stellate cell. Journal of Shanghai Second Medical University, 2008, 20 (2):71-76.
21. Benyon RC,Arthur MJ,.Extracellular matrix degradation and the role of hepatic stellate cells.Semin Liver Dis,2001,21:373-384.
22.徐斌,张振华,黄祝清。血清基质金属蛋白酶及其组织抑制因子与肝纤维化.实用肝脏病杂志,2004,7(1):56-58
23.高润平,齐晓艳。肝纤维化的发生机制与治疗进展。世界华人消化杂志,2006,14(23):2263-2269
24. NIE QH,CHENG YQ,XIE YM. The inhibiting effects of antisense oligonucleotides phosphoroth ioate on gene expression of TMPs in rat liver fibrosis. JG astroentero.2001,7:363.
25.刘海林,王磊。肝纤维化的研究进展。国际消化病杂志。2007,27(4):235-236
1. HammelP,CouvelardA,0’TooleD,RatouisA,SauvanetA,FlejouJF,DegottC,BeighitiJ,BernadesP.VallaD,RusZniewski P,LevyP. Regression of liver fibrosis after biliary drainage in Patients with chronic Pancreatitis and stenosis of the common bile duct. N Engl J Med 2001;344:418-423.
2. Benyon RC, Iredale JP. Is liver fibrosis reversible? Gut 2000; 46: 443-446.
3. Benyon RC, Arthur MJP. Mechanisms of hepatic fibrosis. J Pediatr Gastroente rol Nutr 1998; 27:75-85.
4.姜慧卿,张晓岚。肝纤维化的发生机制。世界华人消化杂志2000;8:687-689
5.张莉娟,王小众,黄月红,陈治新。降钙素基因相关肽、血管紧张素Ⅱ和内皮素在大鼠纤维化中的作用。世界华人消化杂志2001;9:457-459
6. Hayasaka A, Saisho H. Serum markers as tools to monitor liver fibrosis. [J].Digestion, 1998, 59∶381-384.
7.贾继东,尹珊珊。肝纤维化的诊断和治疗进展[J].临床内科杂志,2002,19:244-246
8.张萍。血清LN、CIV、PCⅢ、HA对肝纤维化及肝硬化的诊断价值[J]。中国疗养医学,2002,11(1):50
9.周光德,赵景民。细胞外基质在肝内的代谢与肝纤维化的形成。世界华人消化杂志2002;10(1):57-59
10.蔡卫民,张彬彬,翁红雷等。八项肝纤维化血清标志物比较研究。[J].中华肝脏病杂志2004,12(4)∶219-222.
11.罗瑞红,杨绍基,谢俊强等。肝纤维化血清五项标志物的诊断意义[J].中华肝脏病杂志2001,9(3):148-150
12. Benyon RC, Hovell CJ, Gaca MD, Jones EH, Iredale JP, Arthur MJP. Progelatinas e A is produced and activated by rat hepatic stellate cells and promotes their proliferation. Hepatology 1999;30: 977-986.
13. Nagase H,Woessner JF Jr . Matrix metalloProteinases. J Biol Chem. 1999二30: 21491-21494.
14. MASSOVA IKOTRA LP, FRIDMAN R, MOBASHERY S Matrix metallopro- teinases: structure, evolution, and diversification[ J] FASEB,1998, 12: 1075.
15.朱跃科,王宝恩,申凤俊等。实验性肝纤维化基质金属蛋白酶-2基因表达与酶活性的变化[J]。中华肝脏病杂志,2005,13(7):509-512
16. Pre′aux AM, Mallat A, van Nhieu JT, D′Ortho MP, Hembry RM, Mavier P. Matrix metalloproteinase-2 activation in human hepatic fibrosis regulation by cell-matrix interactions. Hepatology 1999; 30: 944-950.
17. Knittel T, Mehde M, Kobold D, Saile B, Dinter C, Ramadori G. Expressio n patterns of matrix metalloproteinases and their inhibitors in parenchymal and non-parenchymal cells of rat liver: regulation by TNF-αand TGF-β1. J Hepatol 1999; 30:48-60.
18.张亚飞,聂和青,谢玉梅。抑制金属蛋白酶组织抑制因子-2在肝组织中的表达对大鼠肝纤维化的影响[J]。中华肝脏病杂志2006,14:138-140
19. NINOMIYA T, YOON S, NAGANO H,et al. Significance of serum matrix metalloproteinases and their inhibitors on the antifibrogenetic effect of interferorr alfa in chronic hepatitis C patients[J]. Intervirology,2001,44(4): 227-231.
20. GIANNELLI G , BERGAMINI C , MARINOSCI F , et al. Clinical role of MMP-2/TIMP-2 imbalance in hepatocellular carcinoma[J].Int J Cancer,2002, 97(4):425-431.
21. BENYON RC,ARTHUR MJ. Extracellular of matrix degradation and the role of hepatic stellate cells[J].Semin Liver Dis,200l,2l:373-384.
22. Cassiman D, Libbrecht L, Desmet V, Aertsen P, Denef C, Roskams T. Hepatic stellate cell/myofibroblast subpopulations in fibrotic human and rat livers. J Hepatol 2001; 34(suppl,1):20.
23.刘文滨,王吉耀。NF-κB与肝星状细胞凋亡。世界华人消化杂志2001;9:1054-1055
24. ONOM,MLYAMURAM,KYOTANI S,et al. Effect of Shosaiko to ex-tracter on HGF and TGF-beta levels of intraorgans in liver injured rats after partial hepatectomy[J] J Pharm Pharmacol,2000, 52(1): 111.
© 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 |