蛋白芯片技术研究吲哚-3-原醇抗肝纤维化的分子机制

Molecular Mechanisms of Anti-Fibrotic Effect of Indole-3-Carbinol by Protein Array Assays

目的:利用高通量蛋白芯片技术,观察吲哚-3-原醇(I3C)对活化肝星状细胞(HSCs)内多种蛋白表达的影响,初步探讨I3C抗肝纤维化作用的分子机制。方法:培养永生化肝星状细胞株HSC-T6,该细胞具有活化的HSC表型,用100μmol/L的I3C处理细胞24h。高通量蛋白芯片技术观察HSC中与细胞周期、DNA损伤修复、细胞凋亡、骨架蛋白和细胞外基质等相关的主要蛋白的表达变化,分析变化达2倍以上的蛋白分子。结果:I3C可抑制炎症过程,减少HSC活化;阻滞细胞周期的进展,抑制HSC增殖;抑制NF-"B相关信号通路,促进HSC凋亡;调节细胞骨架和细胞外基质的合成与分解,减少肝脏胶原沉积。结论:I3C可通过多途径发挥其抗肝纤维化作用。

Objective：To observe the effects of indole-3-carbinol（I3C）on protein expressions in hepatic stellate cells（HSCs）by using the high-throughput protein arrays.Methods：HSC-T6,an immortalized rat liver stellate cell line,was incubated with 100μmol/L I3C for 24h.Cell cycle,DNA damage repair,cell apoptosis,cytoskeletal and extracellular matrix-related proteins were observed by protein array assays.Accurate differential expression measurements were obtained by selecting genes been up-or down-regulated at least 2-fold in each of the two arrays.Results：Compared with the control,I3Cexerts its anti-fibrotic effects possibly via reducing inflammation process and HSC activation,blocking cell cycle progression and HSC proliferation,inhibiting NF-＂B signal transduction pathway and inducing HSC apoptosis,regulating the balance of extracellular matrix and accelerate collagen degradation.Conclusion：I3C has anti-fibrotic effect on hepatic stellate cells through multiple processes.