ADAMTS-2及TGF-β1在肝硬化组织中的表达
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摘要
目的：检测含Ⅰ型凝血酶敏感蛋白(TSP-1)模体的解整合素样金属蛋白酶(ADAMTS)-2及转化生长因子(TGF)-β1在肝硬化组织中的表达,探讨ADAMTS-2与TGF-β1在肝硬化发生发展中的作用。
方法：收集武汉总医院2010年3月至6月肝硬化门静脉高压症患者的肝组织16例,收集外伤性肝破裂正常肝组织8例作为正常对照组。采用免疫组化及Western blot法检测ADAMTS-2及转化生长因子-β1在肝硬化组织及正常组织中的表达。
结果：免疫组织化学结果显示ADAMTS-2及TGF-β1在组织切片肝硬化组织中表达较正常肝组织中表达明显增高,两者比较差异有统计学意义(P<0.05)。Western blot分析亦显示ADAMTS-2及转化生长因子-β1在肝硬化组织中表达强度明显高于正常组织(P<0.05),而且两者的表达呈正相关(r=0.862,P<0.01)。
结论:ADAMTS-2和TGF-β1在肝硬化组织中表达增高,而且两者间的表达呈正相关。
Objective:To explore the expression and distribution of ADAMTS (a disintegrin and metalloprotease with thrombospondin motif)-2 and TGF (transforming growth factor)-β1 in patients with or without cirrhosis, and to investigate their relations between the two factors.
Methods:The liver tissue from 16 cases with cirrhotic portal hypertensive and 8cases with liver injury were collected from March to June, 2010. Immunohistochemistry and Western blot were applied to detect the protein expression of ADAMTS-2 and TGF-β1.
Results:Immunohistochemistry showed expression level of ADAMTS-2 and TGF-β1 was significantly stronger in liver tissue of cirrhosis group than that in normal group(P<0.05). Western blot also showed expression of ADAMTS-2 and TGF-β1 in liver tissue of cirrhosis group was higher than that in normal group(P<0.05). Furthermore, there was a positive correlation between these two factors(r=0.862, P<0.01).
Conclusion:The expression of ADAMTS-2 and TGF-(31 were significantly increased in cirrhotic liver, while these two factors showed a positive correlation.
方法：收集武汉总医院2010年3月至6月肝硬化门静脉高压症患者的肝组织16例,收集外伤性肝破裂正常肝组织8例作为正常对照组。采用免疫组化及Western blot法检测ADAMTS-2及转化生长因子-β1在肝硬化组织及正常组织中的表达。
结果：免疫组织化学结果显示ADAMTS-2及TGF-β1在组织切片肝硬化组织中表达较正常肝组织中表达明显增高,两者比较差异有统计学意义(P<0.05)。Western blot分析亦显示ADAMTS-2及转化生长因子-β1在肝硬化组织中表达强度明显高于正常组织(P<0.05),而且两者的表达呈正相关(r=0.862,P<0.01)。
结论:ADAMTS-2和TGF-β1在肝硬化组织中表达增高,而且两者间的表达呈正相关。
Objective:To explore the expression and distribution of ADAMTS (a disintegrin and metalloprotease with thrombospondin motif)-2 and TGF (transforming growth factor)-β1 in patients with or without cirrhosis, and to investigate their relations between the two factors.
Methods:The liver tissue from 16 cases with cirrhotic portal hypertensive and 8cases with liver injury were collected from March to June, 2010. Immunohistochemistry and Western blot were applied to detect the protein expression of ADAMTS-2 and TGF-β1.
Results:Immunohistochemistry showed expression level of ADAMTS-2 and TGF-β1 was significantly stronger in liver tissue of cirrhosis group than that in normal group(P<0.05). Western blot also showed expression of ADAMTS-2 and TGF-β1 in liver tissue of cirrhosis group was higher than that in normal group(P<0.05). Furthermore, there was a positive correlation between these two factors(r=0.862, P<0.01).
Conclusion:The expression of ADAMTS-2 and TGF-(31 were significantly increased in cirrhotic liver, while these two factors showed a positive correlation.
引文
[1]王爱民,王宝恩,杨跃伟,等.实验性肝纤维化会属蛋白酶组织抑制因子-Ⅰ基因表达的动态变化.中华实验外科杂志,1998,15：352-353.
[2]赵建学,陆玮婷,郭海燕,等.肝纤维化与细胞因子的研究新进展.中国医药导报,2008,2(5)：19.
[3]Chen YW, Li DG, WU JX, et al. Tetrandrine inhibits activation of rat hepatic stellate cells stimulated by transforming growth factor-beta in vitro via up-regulation of Smad7. J Ethnopharmacol,2005,100(3):299.
[4]Kharbanda KK, Rongers DD, Wyatt TA, et al. Transforming growth factor-beta induces contraction of activated hepatic stellate cells. J Hepatol,2004,41(1):60.
[5]Philips N, Keler T, Gonzalez S. TGF P-like regulation of matrix metalloproteinases by anti-transforming growth factor-β, and anti-transforming growth factor-β1 antibodies in dermal fibroblasts:Implications for wound healing. Wound Repair Regen, 2004,12:53-59.
[6]Bissell DM, Roulot D, George J:Transforming growth factor beta and the liver. Hepatology 2001,34:859-867.
[7]Thenappan A, Li Y, Kitisin K, Rashid A, et al. Role of transforming growth factor beta signaling and expansion of progenitor cells in regenerating liver. Hepatology 2009, 51:1373-1382.
[8]Breitkopf K, Sawitza I, Westhoff JH, Wickert L, Dooley S, Gressner AM: Thrombospondin 1 acts as a strong promoter of transforming growth factor beta effects via two distinct mechanisms in hepatic stellate cells. Gut 2005,54:673-681.
[9]Murphy-Ullrich JE:Thrombospondin-dependent activation of latent TGF-β1-beta in fibrosis and disease. In Cancer Drug Discovery and Development. Humana Press Inc; 2008:549-567:Cancer Drug Discovery And Development:1.
[10]Daniel C, Wiede J, Krutzsch HC, et al. Thrombospondin-1 is a major activator of TGF-β1-beta in fibrotic renal disease in the rat in vivo. Kidney Int 2004,65:459-468.
[11]Crawford SE, Stellmach V, Murphy-Ullrich JE, et al. Thrombospondin-1 is a major activator of TGF-β1-betal in vivo. Cell 1998,93:1159-1170.
[12]El-Youssef M, Mu Y, Huang L, et al. Increased expression of transforming growth factor-betal and thrombospondin-1 in congenital hepatic fibrosis:possible role of the hepatic stellate cell. J Pediatr Gastroenterol Nutr,1999,28(4):p.386-92.
[13]Myung SJ, Yoon JH, Gwak GY, et al. Bile acid-mediated thrombospondin-1 induction in hepatocytes leads to transforming growth factor-beta-dependent hepatic stellate cell activation. Biochem Biophys Res Commun,2007,353(4):p.1091-6.
[14]Elpek GO, Gokhan GA, Bozova S. Thrombospondin-1 expression correlates with angiogenesis in experimental cirrhosis. World J Gastroenterol,2008,14(14):p. 2213-7.
[15]Breitkopf K, Haas S, Wiercinska E, et al.Anti-TGF-β1-beta strategies for the treatment of chronic liver disease. Alcoholism-Clinical And Experimental Research 2005, 29:121S-131S.
[16]李汉军,董超,常实.肝纤维化时转化生长因子-β1对凝血酶致敏感蛋白-1的正向调节作用.中华实验外科杂志,2010,11(27)：1674-1676
[17]Porter S, Clark M, Kevorkian L, et al. The ADAMTS metalloprteinases. J Biochem, 2005,386(15):15-27.
[18]Colige A, Li SW, Sieron AL, et al. cDNA cloning and expression of bovine procollagen I N-proteinase:a new member of the superfamily of zinc-metalloproteinases with binding sites for cells and other matrix components. Proc Natl Acad Sci U S A,1997,94(6):p.2374-9.
[19]Ainsley C Nicholson, Shehre-Banoo Malik, John M Logsdon Jr, et al. Functional evolution of ADAMTS genes:Evidence from analyses of phylogeny and gene organization. BMC Evolutionary Biology,2005,5:11.
[20]Wei-Man Wang, Gaoxiang Ge, Ngee Han Lim, et al. TIMP-3 Inhibits the Procollagen N-proteinase ADAMTS-2. Biochem J.2006,398(Pt 3):515-519.
[21]Schwettmann L, Wehmeier M, Jokovic D, et al. Hepatic expression of A disintegrin and metalloproteinase (ADAM) and ADAMs with thrombospondin motives (ADAM-TS) enzymes in patients with chronic liver diseases. J Hepatol,2008,49(2): p.243-50.
[22]Kesteloot F, Desmouliere A, Leclercq I, et al. ADAM metallopeptidase with thrombospondin type 1 motif 2 inactivation reduces the extent and stability of carbon tetrachloride-induced hepatic fibrosis in mice. Hepatology,2007,46(5):p.1620-31.
[23]Moss ML,Bartsch JW. Therapeutic benefits from targeting of ADAM family members. Biochemistry,2004,43(23):p.7227-35.
[1]Rocks N, Paulissen G,El Hour M, et al. Emerging roles of ADAM and ADAMTS metalloproteinases in cancer [J]. Biochimie,2008,90(2):369-379.
[2]Gilpin BJ, Mattei MG,Engvall E, et al.A novel, secreted form of human ADAM12(meltrin alpha) provokes myogenesis in vivo [J]. J Biol Chem,1998, 273(1):16993-16997.
[3]Lee LW, Peng HC, Ko WC, et al. What have snakes taught us about integrins? [J]. Cell Mol Life Sci,1998,54(12):527-540.
[4]Clissa PB, Lopes-Ferreira M, Della-Casa MS, et al. Importance of jararhagin-like and cysteine-rich domains in the early events of local in amatory response [J]. Toxicon, 2006,47(5):591-596.
[5]Gantus MA, Nasciutti LE, Cruz CM, et al. Modulation of extracellularmatrix components by metalloproteinases and their tissue inhibitors during degeneration and regeneration of rat sural nerve [J]. Brain Res,2006,2233(1):36-46.
[6]Dawson DW, Pearce SF, Zhong R, et al. CD36 mediates the in vitro inhibitory effects of thrombospondin-1 on endothelial cells [J]. J Cell Biol,1997,138(3):707-717.
[7]Porter S, Clark M, Kevorkian L, et al. The ADAMTS metalloprteinases [J]. J Biochem, 2005,386(15):15-27.
[8]Reiss K, Saftig P. The "A Disintegrin And Metalloprotease"(ADAM) family of sheddases:Physiological and cellular functions [J]. Semin Cell Devt Biol,2009, 20(2):126-137.
[9]Porter S, Clark IM, Kevorkian L, Edwards DR. The ADAMTS metalloproteinases. Biochem J,2005,386(Pt 1):p.15-27.
[10]Colige A, Li SW, Sieron AL, Nusgens BV, Prockop DJ, Lapiere CM. cDNA cloning and expression of bovine procollagen I N-proteinase:a new member of the superfamily of zinc-metalloproteinases with binding sites for cells and other matrix components. Proc Natl Acad Sci U S A,1997,94(6):p.2374-9.
[11]Li SW, Arita M, Fertala A, et al. Transgenic mice with inactive alleles for procollagen N-proteinase (ADAMTS-2) develop fragile skin and male sterility. Biochem J,2001, 355:271-278.
[12]Colige A, Sieron AL, Li SW, et al. Human Ehlers-Danlos syndrome type VII C and bovine dermatosparaxis are caused by mutations in the procollagen I N-proteinase gene. Am J Hum Genet.1999,65:308-317.
[13]Schwettmann L, Wehmeier M, Jokovic D, Aleksandrova K, Brand K, Manns MP, Lichtinghagen R, Bahr MJ. Hepatic expression of A disintegrin and metalloproteinase (ADAM) and ADAMs with thrombospondin motives (ADAM-TS) enzymes in patients with chronic liver diseases [J]. J Hepatol,2008,49(2):p.243-50.
[14]Kesteloot F, Desmouliere A, Leclercq I, Thiry M, Arrese JE, Prockop DJ, Lapiere CM, Nusgens BV, Colige A. ADAM metallopeptidase with thrombospondin type 1 motif 2 inactivation reduces the extent and stability of carbon tetrachloride-induced hepatic fibrosis in mice. Hepatology [J],2007,46(5):p.1620-31.
[15]Colige A, Ruggiero F, Vandenberghe I, et al. Domains and maturation processes that regulate the activity of ADAMTS-2, a metalloproteinase cleaving the aminopropeptide of fibrillar procollagens type Ⅰ-Ⅲ and Ⅴ [J]. J Biol Chem, 2005,280:34397-34408.
[16]Ainsley C Nicholson, Shehre-Banoo Malik, John M Logsdon Jr, et al. Functional evolution of ADAMTS genes:Evidence from analyses of phylogeny and gene organization[J]. BMC Evolutionary Biology,2005,5:11.
[17]Wei-Man Wang, Gaoxiang Ge, Ngee Han Lim, et al. TIMP-3 Inhibits the Procollagen N-proteinase ADAMTS-2[J]. Biochem J.2006,398(Pt 3):515-519.
[18]Moss ML,Bartsch JW. Therapeutic benefits from targeting of ADAM family members[J]. Biochemistry,2004,43(23):p.7227-35.
[2]赵建学,陆玮婷,郭海燕,等.肝纤维化与细胞因子的研究新进展.中国医药导报,2008,2(5)：19.
[3]Chen YW, Li DG, WU JX, et al. Tetrandrine inhibits activation of rat hepatic stellate cells stimulated by transforming growth factor-beta in vitro via up-regulation of Smad7. J Ethnopharmacol,2005,100(3):299.
[4]Kharbanda KK, Rongers DD, Wyatt TA, et al. Transforming growth factor-beta induces contraction of activated hepatic stellate cells. J Hepatol,2004,41(1):60.
[5]Philips N, Keler T, Gonzalez S. TGF P-like regulation of matrix metalloproteinases by anti-transforming growth factor-β, and anti-transforming growth factor-β1 antibodies in dermal fibroblasts:Implications for wound healing. Wound Repair Regen, 2004,12:53-59.
[6]Bissell DM, Roulot D, George J:Transforming growth factor beta and the liver. Hepatology 2001,34:859-867.
[7]Thenappan A, Li Y, Kitisin K, Rashid A, et al. Role of transforming growth factor beta signaling and expansion of progenitor cells in regenerating liver. Hepatology 2009, 51:1373-1382.
[8]Breitkopf K, Sawitza I, Westhoff JH, Wickert L, Dooley S, Gressner AM: Thrombospondin 1 acts as a strong promoter of transforming growth factor beta effects via two distinct mechanisms in hepatic stellate cells. Gut 2005,54:673-681.
[9]Murphy-Ullrich JE:Thrombospondin-dependent activation of latent TGF-β1-beta in fibrosis and disease. In Cancer Drug Discovery and Development. Humana Press Inc; 2008:549-567:Cancer Drug Discovery And Development:1.
[10]Daniel C, Wiede J, Krutzsch HC, et al. Thrombospondin-1 is a major activator of TGF-β1-beta in fibrotic renal disease in the rat in vivo. Kidney Int 2004,65:459-468.
[11]Crawford SE, Stellmach V, Murphy-Ullrich JE, et al. Thrombospondin-1 is a major activator of TGF-β1-betal in vivo. Cell 1998,93:1159-1170.
[12]El-Youssef M, Mu Y, Huang L, et al. Increased expression of transforming growth factor-betal and thrombospondin-1 in congenital hepatic fibrosis:possible role of the hepatic stellate cell. J Pediatr Gastroenterol Nutr,1999,28(4):p.386-92.
[13]Myung SJ, Yoon JH, Gwak GY, et al. Bile acid-mediated thrombospondin-1 induction in hepatocytes leads to transforming growth factor-beta-dependent hepatic stellate cell activation. Biochem Biophys Res Commun,2007,353(4):p.1091-6.
[14]Elpek GO, Gokhan GA, Bozova S. Thrombospondin-1 expression correlates with angiogenesis in experimental cirrhosis. World J Gastroenterol,2008,14(14):p. 2213-7.
[15]Breitkopf K, Haas S, Wiercinska E, et al.Anti-TGF-β1-beta strategies for the treatment of chronic liver disease. Alcoholism-Clinical And Experimental Research 2005, 29:121S-131S.
[16]李汉军,董超,常实.肝纤维化时转化生长因子-β1对凝血酶致敏感蛋白-1的正向调节作用.中华实验外科杂志,2010,11(27)：1674-1676
[17]Porter S, Clark M, Kevorkian L, et al. The ADAMTS metalloprteinases. J Biochem, 2005,386(15):15-27.
[18]Colige A, Li SW, Sieron AL, et al. cDNA cloning and expression of bovine procollagen I N-proteinase:a new member of the superfamily of zinc-metalloproteinases with binding sites for cells and other matrix components. Proc Natl Acad Sci U S A,1997,94(6):p.2374-9.
[19]Ainsley C Nicholson, Shehre-Banoo Malik, John M Logsdon Jr, et al. Functional evolution of ADAMTS genes:Evidence from analyses of phylogeny and gene organization. BMC Evolutionary Biology,2005,5:11.
[20]Wei-Man Wang, Gaoxiang Ge, Ngee Han Lim, et al. TIMP-3 Inhibits the Procollagen N-proteinase ADAMTS-2. Biochem J.2006,398(Pt 3):515-519.
[21]Schwettmann L, Wehmeier M, Jokovic D, et al. Hepatic expression of A disintegrin and metalloproteinase (ADAM) and ADAMs with thrombospondin motives (ADAM-TS) enzymes in patients with chronic liver diseases. J Hepatol,2008,49(2): p.243-50.
[22]Kesteloot F, Desmouliere A, Leclercq I, et al. ADAM metallopeptidase with thrombospondin type 1 motif 2 inactivation reduces the extent and stability of carbon tetrachloride-induced hepatic fibrosis in mice. Hepatology,2007,46(5):p.1620-31.
[23]Moss ML,Bartsch JW. Therapeutic benefits from targeting of ADAM family members. Biochemistry,2004,43(23):p.7227-35.
[1]Rocks N, Paulissen G,El Hour M, et al. Emerging roles of ADAM and ADAMTS metalloproteinases in cancer [J]. Biochimie,2008,90(2):369-379.
[2]Gilpin BJ, Mattei MG,Engvall E, et al.A novel, secreted form of human ADAM12(meltrin alpha) provokes myogenesis in vivo [J]. J Biol Chem,1998, 273(1):16993-16997.
[3]Lee LW, Peng HC, Ko WC, et al. What have snakes taught us about integrins? [J]. Cell Mol Life Sci,1998,54(12):527-540.
[4]Clissa PB, Lopes-Ferreira M, Della-Casa MS, et al. Importance of jararhagin-like and cysteine-rich domains in the early events of local in amatory response [J]. Toxicon, 2006,47(5):591-596.
[5]Gantus MA, Nasciutti LE, Cruz CM, et al. Modulation of extracellularmatrix components by metalloproteinases and their tissue inhibitors during degeneration and regeneration of rat sural nerve [J]. Brain Res,2006,2233(1):36-46.
[6]Dawson DW, Pearce SF, Zhong R, et al. CD36 mediates the in vitro inhibitory effects of thrombospondin-1 on endothelial cells [J]. J Cell Biol,1997,138(3):707-717.
[7]Porter S, Clark M, Kevorkian L, et al. The ADAMTS metalloprteinases [J]. J Biochem, 2005,386(15):15-27.
[8]Reiss K, Saftig P. The "A Disintegrin And Metalloprotease"(ADAM) family of sheddases:Physiological and cellular functions [J]. Semin Cell Devt Biol,2009, 20(2):126-137.
[9]Porter S, Clark IM, Kevorkian L, Edwards DR. The ADAMTS metalloproteinases. Biochem J,2005,386(Pt 1):p.15-27.
[10]Colige A, Li SW, Sieron AL, Nusgens BV, Prockop DJ, Lapiere CM. cDNA cloning and expression of bovine procollagen I N-proteinase:a new member of the superfamily of zinc-metalloproteinases with binding sites for cells and other matrix components. Proc Natl Acad Sci U S A,1997,94(6):p.2374-9.
[11]Li SW, Arita M, Fertala A, et al. Transgenic mice with inactive alleles for procollagen N-proteinase (ADAMTS-2) develop fragile skin and male sterility. Biochem J,2001, 355:271-278.
[12]Colige A, Sieron AL, Li SW, et al. Human Ehlers-Danlos syndrome type VII C and bovine dermatosparaxis are caused by mutations in the procollagen I N-proteinase gene. Am J Hum Genet.1999,65:308-317.
[13]Schwettmann L, Wehmeier M, Jokovic D, Aleksandrova K, Brand K, Manns MP, Lichtinghagen R, Bahr MJ. Hepatic expression of A disintegrin and metalloproteinase (ADAM) and ADAMs with thrombospondin motives (ADAM-TS) enzymes in patients with chronic liver diseases [J]. J Hepatol,2008,49(2):p.243-50.
[14]Kesteloot F, Desmouliere A, Leclercq I, Thiry M, Arrese JE, Prockop DJ, Lapiere CM, Nusgens BV, Colige A. ADAM metallopeptidase with thrombospondin type 1 motif 2 inactivation reduces the extent and stability of carbon tetrachloride-induced hepatic fibrosis in mice. Hepatology [J],2007,46(5):p.1620-31.
[15]Colige A, Ruggiero F, Vandenberghe I, et al. Domains and maturation processes that regulate the activity of ADAMTS-2, a metalloproteinase cleaving the aminopropeptide of fibrillar procollagens type Ⅰ-Ⅲ and Ⅴ [J]. J Biol Chem, 2005,280:34397-34408.
[16]Ainsley C Nicholson, Shehre-Banoo Malik, John M Logsdon Jr, et al. Functional evolution of ADAMTS genes:Evidence from analyses of phylogeny and gene organization[J]. BMC Evolutionary Biology,2005,5:11.
[17]Wei-Man Wang, Gaoxiang Ge, Ngee Han Lim, et al. TIMP-3 Inhibits the Procollagen N-proteinase ADAMTS-2[J]. Biochem J.2006,398(Pt 3):515-519.
[18]Moss ML,Bartsch JW. Therapeutic benefits from targeting of ADAM family members[J]. Biochemistry,2004,43(23):p.7227-35.