| 组织蛋白酶B在肝星状细胞HSC-T6中的动态表达和意义 |
| 投稿时间：2011-06-23 修订日期：2011-07-04 点此下载全文 |
| 引用本文：李春艳,陈永平,申春燕.组织蛋白酶B在肝星状细胞HSC-T6中的动态表达和意义[J].医学研究杂志,2012,41(3):36-41 |
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| 基金项目:浙江省自然基金资助项目(Y207464) |
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| 中文摘要: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-β mRNA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±047）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±008）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。 |
| 中文关键词:组织蛋白酶B 肝星状细胞 肝纤维化 |
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| Dynamic Expression of Cathepsin B in Hepatic Stellate Cells (HSC-T6) and Its Significance |
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| Abstract:ObjectiveTo observe the expression of cathepsin B in hepatic stellate cells (HSC-T6) and explore its significance of Cathepsin B in liver fibrosis MethodsWe 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 cathepsin 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. ResultsZ-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 significant(t=-6.31,P<0.05).The variation of α-SMA expression was parallel with the variation of cathepsin B expression 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 significant(t=-6.18,P<0.05).The results of RT-PCR showed that the cathepsin B mRNA expression of inhibitor group（200±0.11）decreased compared to the control group（2.24±0.47）(t=-3.41,P<0.05),and theα-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).ConclusionCathepsin B could inhibit HSC proliferation and transdifferentiation, and result in the downregulation of liver fibrosis indexs, such as TGF-βandα-SMA. So, cathepsin B may be a new potencial therapeutic targets for the treatment of liver fibrosis in future. |
| keywords:Cathepsin B Hepatic stellate cell Liver fibrosis |
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