组织蛋白酶B在肝星状细胞HSC-T6中的动态表达和意义  

Dynamic Expression of Cathepsin B in Hepatic Stellate Cells(HSC-T6) and Its Significance

作　　者：李春艳[1] 陈永平[1] 申春燕[1] 

机构地区：[1]温州医学院附属第一医院感染内科,325000

出　　处：《医学研究杂志》2012年第3期36-41,共6页Journal of Medical Research

基　　金：浙江省自然基金资助项目(Y207464)

摘　　要：目的观察组织蛋白酶B在肝星状细胞(hepatic stellate cell,HSC-T6)中的表达并探讨其在肝纤维化过程中的意义。方法向HSC-T6细胞中加入组织蛋白酶B抑制剂(Z-FA-FMK),作用12、24、36、48h后,用倒置显微镜分别观察各时间点细胞生长状态的差异并拍照。分别提取Z-FA-FMK作用前后12、24、36、48h的蛋白和RNA,用Western blotting法检测各时间点组织蛋白酶B(cathepsin B)和α-SMA的表达量,用反转录定量PCR(RT-PCR)方法测定各时间点cathepsin B mRNA、TGF-βmRNA和α-SMA mRNA的表达。统计学处理采用配对t检验和Pearson直线相关分析。结果显微镜观察Z-FA-FMK作用后细胞的增殖分化作用减慢。Western blotting结果显示,对照组和抑制剂组中cathepsin B的表达量随时间的延长而增加,抑制剂组cathepsin B的表达量(0.49±0.04)较对照组(0.68±0.09)有所减少,差异具有统计学意义(t=-6.31,P<0.05);α-SMA表达量的变化与cathepsin B的变化平行,抑制剂组α-SMA表达量(0.64±0.03)较对照组(0.79±0.01)有所减少,差异具有统计学意义(t=-6.18,P<0.05)。RT-PCR结果显示抑制剂组cathepsin B mRNA表达量(2.00±0.11)较对照组(2.24±0.47)减少,差异均具有统计学意义(t=-3.41,P<0.05);抑制剂组α-SMA mRNA的表达量(0.40±0.06)较对照组(0.81±0.08)减少,差异具有统计学意义(t=-4.28,P<0.05);抑制剂组TGF-βmRNA的表达量(1.43±0.16)较对照组(1.81±0.21)减少,差异均具有统计学意义(t=6.44,P<0.05)。结论组织蛋白酶B可抑制HSC的增殖和转分化,同时可降低肝纤维化指标如TGF-β和α-SMA的表达量,因此,组织蛋白酶B可作为今后治疗肝纤维化的治疗靶点。Objective To observe the expression of cathepsin B in hepatic stellate cells （HSC - T6） and explore its significance of Cathepsin B in liver fibrosis Methods We put Z - FA - FMK into HSC - T6 cell medium of the inhibitor group to observe and compare the HSCs form changes at 12h,24h,36h,48h by microscopy. We extracted the protein and mRNA of HSCs at different time -points during culture,and detected the expression of eathepsin B, TGF -β and α- SMA both at level of mRNA and protein respectively with reverse transcription -polymerase chain reaction（ RT-PCR） and Western blotting. The statistical analysis was done using paired t test and Pearson liner correlation analysis. Results Z - FA - FMK decreased HSCs proliferation and differentiation was detected by microscopy. The results of Western blot showed that the expression of cathepsin B increased with the extension of time both in control group and inhibitor group. Compared to the control group（0.68 ± 0.09 ） , cathepsin B expression of inhibitor group （0.49 ±0. 04） decreased, and there was statistically signifieant（t = -6.31 ,P 〈0.05）. The variation of α-SMA expression was parallel with the variation of cathepsin B expres- sion and α-SMA expression of inhibitor group（0.64 ± 0. 03 ） decreased compared to the control group（0.79± 0. 01 ） , and there was statistically signifieant（t = -6. 18 ,P 〈 0.05 ）. The results of RT-PCR showed that the cathepsin B mRNA expression of inhibitor group （ 2.00 ±0. 11 ） decreased compared to the control group （ 2.24 ± 0.47 ） （ t = - 3.41, P 〈 0.05 ） , and the α-SMA mRNA expression of inhibitor group （0.40±0.06） decreased compared to the control group（0. 81±0. 08 ） （ t = - 4.28, P 〈0. 05 ） , and the TGF - βmRNA expression of inhibitor group（1.43 ±0.16）decreased compared to the control group（1.81 ±0.21）（t=6.44,P〈0.05）. Conclusion Cathepsin B could inhibit HSC proliferation and transdifferentiation, and result in the downregulation of liver fibrosis

关 键 词：组织蛋白酶B 肝星状细胞 肝纤维化 

分 类 号：R735.2[医药卫生—肿瘤]