MMP-7在胆道闭锁肝纤维化患者中的表达及临床意义
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摘要
目的研究基质金属蛋白酶7(MMP-7)与胆道闭锁(BA)患者肝纤维化分级、胆管反应程度之间的关系,探索其在肝纤维化进程中的作用及临床意义。方法选取2017年6月—2018年4月我院收治的15例胆道闭锁患儿行Kasai手术时的肝活检组织(G1组),另选取2015年1月—2016年1月因行Kasai术后肝脏功能衰竭而行肝移植术的病肝组织41例(移植组),将移植组按Kasai手术与肝移植的间隔时间分为2组:G2组(28例,间隔时间<2年)和G3组(13例,间隔时间≥2年)。采用HE染色观察3组患者肝脏纤维化程度,免疫组化染色检测MMP-7和CK-19蛋白在肝脏中的表达情况并进行半定量分析。结果 (1)免疫组化结果显示,MMP-7主要表达于胆管上皮细胞、增生的胆管、汇管区周围肝细胞,随着肝纤维化程度进展,胆管细胞及肝细胞MMP-7表达强度增加,且胆管细胞比肝细胞表达更为强烈。MMP-7在G1～G3组中的表达水平分别为(0.068±0.017)vs.(0.093±0.017)vs.(0.084±0.016),差异有统计学意义(F=10.541,P<0.001),CK-19在3组的表达水平分别为(0.119±0.036)vs.(0.157±0.040)vs.(0.110±0.044),差异有统计学意义(F=7.296,P<0.001),G2组MMP-7、CK-19蛋白表达水平明显高于其余2组(P<0.05),G1组与G3组之间差异无统计学意义(P>0.05)。(2)3组的肝纤维化程度分别为2(2,3)vs. 4(3,4)vs. 4(2,4)(H=17.785,P<0.001),G2组纤维化程度明显高于其余2组,差异具有统计学意义(P<0.017)。(3)相关性检验示:MMP-7蛋白表达量与肝纤维化程度呈弱正相关(r_s=0.609,P<0.001),但与胆管反应程度(CK-19)无明显线性相关性(r=0.007,P=0.962)。结论MMP-7在BA患儿Kasai术后肝组织中高表达,并且随肝纤维化程度加重而表达升高,其可能参与并促进了BA肝纤维化进程,提示自体肝生存不佳。
Objective To investigate the relationship of matrix metalloproteinase-7(MMP-7) in liver tissue of biliary atresia(BA) with liver fibrosis and bile duct reaction, and explore the role of MMP-7 in the process of liver fibrosis and its clinical significance. Methods Data of 15 BA patients with liver biopsy in Kasai operation in our hospital from June 2017 to April 2018 were collected(G1 group), and data of 41 patients with liver biopsy who underwent liver transplantation after Kasai operation from January 2015 to January 2016 were enrolled as transplantation group. The transplantation group was divided into two groups according to the interval time between Kasai operation and liver transplantation: G2 group(n=28, the interval time was less than 2 years) and G3 group(n=13, the interval time was more than 2 years) Hematoxylin-Eosin(HE)staining was used to observe the degree of liver fibrosis. Immunohistochemical staining was used to detect the expressions of MMP-7 and CK-19. Results(1) mmunohistochemical results showed that MMP-7 was mainly expressed in bile duct epithelial cells, hyperplastic bile ducts and hepatocytes around the portal area. With the progress of hepatic fibrosis, the expressions of MMP-7 were increased in biliary duct cells and hepatocytes, and which were strongly expressed in biliary duct cells than those of hepatocytes. Semi-quantitative analysis showed that the expression levels of MMP-7 were(0.068±0.017) vs.(0.093±0.017) vs.(0.084±0.016) in the three groups(F=10.541, P< 0.001). CK-19 staining showed that bile duct reactions of three groups were(0.119±0.036) vs.(0.157±0.040) vs.(0.110±0.044)(F=7.296,P < 0.001). The expression levels of MMP-7 and CK-19 protein were significantly higher in G2 group than those of other two groups(P < 0.05). There was no significant difference between G1 and G3 groups(P > 0.05).(2) The liver fibrosis in the three groups were 2(2, 3) vs. 4(3, 4)vs. 4(2, 4)(H= 17.785, P < 0.001), and the fibrosis was significantly higher in the G2 group than that of other two groups(P <0.017).(3) Correlation test showed that the expression of MMP-7 protein was positively correlated with the degree of liver fibrosis(r_s=0.609, P< 0.001), but there was no correlation with bile duct reaction(r=0.007, P=0.962). Conclusion The expression of MMP-7 protein is highly expressed in liver tissue after Kasai operation, and it increases with the severity of liver fibrosis. MMP-7 may participate in the process of BA liver fibrosis and suggest that the survival of autologous liver is poor.
Objective To investigate the relationship of matrix metalloproteinase-7(MMP-7) in liver tissue of biliary atresia(BA) with liver fibrosis and bile duct reaction, and explore the role of MMP-7 in the process of liver fibrosis and its clinical significance. Methods Data of 15 BA patients with liver biopsy in Kasai operation in our hospital from June 2017 to April 2018 were collected(G1 group), and data of 41 patients with liver biopsy who underwent liver transplantation after Kasai operation from January 2015 to January 2016 were enrolled as transplantation group. The transplantation group was divided into two groups according to the interval time between Kasai operation and liver transplantation: G2 group(n=28, the interval time was less than 2 years) and G3 group(n=13, the interval time was more than 2 years) Hematoxylin-Eosin(HE)staining was used to observe the degree of liver fibrosis. Immunohistochemical staining was used to detect the expressions of MMP-7 and CK-19. Results(1) mmunohistochemical results showed that MMP-7 was mainly expressed in bile duct epithelial cells, hyperplastic bile ducts and hepatocytes around the portal area. With the progress of hepatic fibrosis, the expressions of MMP-7 were increased in biliary duct cells and hepatocytes, and which were strongly expressed in biliary duct cells than those of hepatocytes. Semi-quantitative analysis showed that the expression levels of MMP-7 were(0.068±0.017) vs.(0.093±0.017) vs.(0.084±0.016) in the three groups(F=10.541, P< 0.001). CK-19 staining showed that bile duct reactions of three groups were(0.119±0.036) vs.(0.157±0.040) vs.(0.110±0.044)(F=7.296,P < 0.001). The expression levels of MMP-7 and CK-19 protein were significantly higher in G2 group than those of other two groups(P < 0.05). There was no significant difference between G1 and G3 groups(P > 0.05).(2) The liver fibrosis in the three groups were 2(2, 3) vs. 4(3, 4)vs. 4(2, 4)(H= 17.785, P < 0.001), and the fibrosis was significantly higher in the G2 group than that of other two groups(P <0.017).(3) Correlation test showed that the expression of MMP-7 protein was positively correlated with the degree of liver fibrosis(r_s=0.609, P< 0.001), but there was no correlation with bile duct reaction(r=0.007, P=0.962). Conclusion The expression of MMP-7 protein is highly expressed in liver tissue after Kasai operation, and it increases with the severity of liver fibrosis. MMP-7 may participate in the process of BA liver fibrosis and suggest that the survival of autologous liver is poor.
引文
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[2]詹江华,冯杰雄.胆道闭锁何时完成Kasai手术[J].中华小儿外科杂志,2016,37(5):321-326.Zhan JH,Feng JX.When compete Kasai of biliary atresia?[J].Chin J Pediatr Surg,2016,37(5):321-326.doi:10.3760/cma.j.issn.0253-3006.2016.05.001.
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[5]董淳强,董昆,杨体泉.基质金属蛋白酶2在胆道闭锁患儿肝组织中的表达[J].中华实用儿科临床杂志,2013,28(23):1801-1804.Dong CQ,Dong K,Yang TQ.Expression of matrix metalloproteinase-2 in liver tissues of children with biliary atresia[J].Chin J Appl Clin Pediatr,2013,28(23):1801-1804.doi:10.3760/cma.j.issn.2095-428X.2013.23.014.
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[7]Baba H,Ohtsuka Y,Fujii T,et al.Immunological investigation of the hepatic tissue from infants with biliary atresia[J].Pediatr Surg Int,2009,25(2):157-162.doi:10.1007/s00383-008-2311-9.
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[9]高伟.小儿肝移植的指征[J].临床小儿外科杂志,2017,16(2):121-126.Gao W.Indication of pediatric liver transplantation[J].JClin Ped Sur,2017,16(2):121-126.doi:10.3969/j.issn.1671-6353.2017.02.005.
[10]Fumino S,Higuchi K,Aoi S,et al.Clinical analysis of liver fibrosis in choledochal cyst[J].Pediatr Surg Int,2013,29(11):1097-1102.doi:10.1007/s00383-013-3368-7.
[11]Davenport M.Biliary atresia:from Australia to the zebrafish[J].JPediatr Surg,2016,51(2):200-205.doi:10.1016/j.jpedsurg.2015.10.058.
[12]张树建,陈扬,高婷,等.RhoA、Rac1及Cdc42在胆道闭锁肝纤维化中的作用[J].中华小儿外科杂志,2017,38(11):816-821.Zhang SJ,Chen Y,Gao T,et al.Effects of RhoA,Rac1 and Cdc42on liver fibrosis in patients with biliary atresia[J].Chin J Pediatr Surg,2017,38(11):816-821.doi:10.3760/cma.j.issn.0253-3006.2017.11.003.
[13]Zhang Q,Liu S,Parajuli KR,et al.Interleukin-17 promotes prostate cancer via MMP7-induced epithelial-to-mesenchymal transition[J].Oncogene,2017,36(5):687-699.doi:10.1038/onc.2016.240.
[14]Irvine KM,Wockner LF,Hoffmann L,et al.Multiplex serum protein analysis identifies novel biomarkers of advanced fibrosis in patients with chronic liver disease with the potential to improve diagnostic accuracy of established biomarkers[J].PLoS One,2016,11(11):e0167001.doi:10.1371/journal.pone.0167001.
[15]Huang CC,Chuang JH,Chou MH,et al.Matrilysin(MMP-7)is a major matrix metalloproteinase upregulated in biliary atresiaassociated liver fibrosis[J].Mod Pathol,2005,18(7):941-950.doi:10.1038/modpathol.3800374.
[16]刘亭睁,张志波.金属蛋白酶7及金属蛋白酶19在胆道闭锁幼鼠动物模型肝脏组织中的表达[J].中国医科大学学报,2017,46(12):1071-1081.Liu TZ,Zhang ZB.Expression of matrix metalloproteinase-7 and-19 in liver tissues of rats with biliary atresia[J].Journal of China Medical University,2017,46(12):1071-1081.doi:10.12007/j.issn.0258-4646.2017.12.004.
[17]Kerola A,Lampela H,Lohi J,et al.Increased MMP-7 expression in biliary epithelium and serum underpins native liver fibrosis aftersuccessful portoenterostomy in biliary atresia[J].J Pathol Clin Res,2016,2(3):187-198.doi:10.1002/cjp2.50.
[18]Nadler EP,Li X,Onyedika E,et al.Differential expression of hepatic fibrosis mediators in sick and spontaneously recovered mice with experimental biliary atresia[J].J Surg Res,2010,159(2):611-617.doi:10.1016/j.jss.2009.10.038.
[19]丁美云,詹江华,赵丽,等.TGF-β1、Smad2在胆道闭锁肝纤维化中的作用[J].天津医药,2016,44(7):810-813.Ding MY,Zhan JH,Zhao L,et al.The effects of TGF-β1 and Smad2 on liver fibrosis of biliary atresia[J].Tianjin Med J,2016,44(7):810-813.doi:10.11958/20150242.
[20]Xiao Y,Zhou Y,Chen Y,et al.The expression of epithelialmesenchymal transition-related proteins in biliary epithelial cells is associated with liver fibrosis in biliary atresia[J].Pediatr Res,2015,77(2):310-315.doi:10.1038/pr.2014.181.
[2]詹江华,冯杰雄.胆道闭锁何时完成Kasai手术[J].中华小儿外科杂志,2016,37(5):321-326.Zhan JH,Feng JX.When compete Kasai of biliary atresia?[J].Chin J Pediatr Surg,2016,37(5):321-326.doi:10.3760/cma.j.issn.0253-3006.2016.05.001.
[3]Iredale JP,Thompson A,Henderson NC.Extracellular matrix degradation in liver fibrosis:Biochemistry and regulation[J].Biochim Biophys Acta,2013,1832(7):876-883.doi:10.1016/j.bbadis.2012.11.002.
[4]Duarte S,Baber J,Fujii T,et al.Matrix metalloproteinases in liver injury,repair and fibrosis[J].Matrix Biol,2015,44(46):147-156.doi:10.1016/j.matbio.2015.01.004.
[5]董淳强,董昆,杨体泉.基质金属蛋白酶2在胆道闭锁患儿肝组织中的表达[J].中华实用儿科临床杂志,2013,28(23):1801-1804.Dong CQ,Dong K,Yang TQ.Expression of matrix metalloproteinase-2 in liver tissues of children with biliary atresia[J].Chin J Appl Clin Pediatr,2013,28(23):1801-1804.doi:10.3760/cma.j.issn.2095-428X.2013.23.014.
[6]Honsawek S,Praianantathavorn K,Chongsrisawat V,et al.High serum matrix metalloproteinase-3 and liver stiffness in postoperative biliary atresia[J].Pediatr Surg Int,2011,27(7):681-687.doi:10.1007/s00383-010-2816-x.
[7]Baba H,Ohtsuka Y,Fujii T,et al.Immunological investigation of the hepatic tissue from infants with biliary atresia[J].Pediatr Surg Int,2009,25(2):157-162.doi:10.1007/s00383-008-2311-9.
[8]Lertudomphonwanit C,Mourya R,Fei L,et al.Large-scale proteomics identifies MMP-7 as a sentinel of epithelial injury and of biliary atresia[J].Sci Transl Med,2017,9(417).pii:eaan8462.doi:10.1126/scitranslmed.aan8462.
[9]高伟.小儿肝移植的指征[J].临床小儿外科杂志,2017,16(2):121-126.Gao W.Indication of pediatric liver transplantation[J].JClin Ped Sur,2017,16(2):121-126.doi:10.3969/j.issn.1671-6353.2017.02.005.
[10]Fumino S,Higuchi K,Aoi S,et al.Clinical analysis of liver fibrosis in choledochal cyst[J].Pediatr Surg Int,2013,29(11):1097-1102.doi:10.1007/s00383-013-3368-7.
[11]Davenport M.Biliary atresia:from Australia to the zebrafish[J].JPediatr Surg,2016,51(2):200-205.doi:10.1016/j.jpedsurg.2015.10.058.
[12]张树建,陈扬,高婷,等.RhoA、Rac1及Cdc42在胆道闭锁肝纤维化中的作用[J].中华小儿外科杂志,2017,38(11):816-821.Zhang SJ,Chen Y,Gao T,et al.Effects of RhoA,Rac1 and Cdc42on liver fibrosis in patients with biliary atresia[J].Chin J Pediatr Surg,2017,38(11):816-821.doi:10.3760/cma.j.issn.0253-3006.2017.11.003.
[13]Zhang Q,Liu S,Parajuli KR,et al.Interleukin-17 promotes prostate cancer via MMP7-induced epithelial-to-mesenchymal transition[J].Oncogene,2017,36(5):687-699.doi:10.1038/onc.2016.240.
[14]Irvine KM,Wockner LF,Hoffmann L,et al.Multiplex serum protein analysis identifies novel biomarkers of advanced fibrosis in patients with chronic liver disease with the potential to improve diagnostic accuracy of established biomarkers[J].PLoS One,2016,11(11):e0167001.doi:10.1371/journal.pone.0167001.
[15]Huang CC,Chuang JH,Chou MH,et al.Matrilysin(MMP-7)is a major matrix metalloproteinase upregulated in biliary atresiaassociated liver fibrosis[J].Mod Pathol,2005,18(7):941-950.doi:10.1038/modpathol.3800374.
[16]刘亭睁,张志波.金属蛋白酶7及金属蛋白酶19在胆道闭锁幼鼠动物模型肝脏组织中的表达[J].中国医科大学学报,2017,46(12):1071-1081.Liu TZ,Zhang ZB.Expression of matrix metalloproteinase-7 and-19 in liver tissues of rats with biliary atresia[J].Journal of China Medical University,2017,46(12):1071-1081.doi:10.12007/j.issn.0258-4646.2017.12.004.
[17]Kerola A,Lampela H,Lohi J,et al.Increased MMP-7 expression in biliary epithelium and serum underpins native liver fibrosis aftersuccessful portoenterostomy in biliary atresia[J].J Pathol Clin Res,2016,2(3):187-198.doi:10.1002/cjp2.50.
[18]Nadler EP,Li X,Onyedika E,et al.Differential expression of hepatic fibrosis mediators in sick and spontaneously recovered mice with experimental biliary atresia[J].J Surg Res,2010,159(2):611-617.doi:10.1016/j.jss.2009.10.038.
[19]丁美云,詹江华,赵丽,等.TGF-β1、Smad2在胆道闭锁肝纤维化中的作用[J].天津医药,2016,44(7):810-813.Ding MY,Zhan JH,Zhao L,et al.The effects of TGF-β1 and Smad2 on liver fibrosis of biliary atresia[J].Tianjin Med J,2016,44(7):810-813.doi:10.11958/20150242.
[20]Xiao Y,Zhou Y,Chen Y,et al.The expression of epithelialmesenchymal transition-related proteins in biliary epithelial cells is associated with liver fibrosis in biliary atresia[J].Pediatr Res,2015,77(2):310-315.doi:10.1038/pr.2014.181.