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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 effect of indole-3-propenol (I3C) on the expression of multiple proteins in activated hepatic stellate cells (HSCs) by high-throughput protein microarray and to explore the molecular mechanism of I3C against hepatic fibrosis. Methods: Immortalized HSC-T6 cells were cultured and activated HSC phenotype. The cells were treated with 100μmol / L I3C for 24 hours. High-throughput protein microarray technique was used to observe the expression of major proteins involved in cell cycle, DNA damage repair, apoptosis, scaffold protein and extracellular matrix in HSC, and the protein molecules with more than 2-fold changes were analyzed. Results: I3C could inhibit the inflammatory process, reduce the activation of HSC, arrest the progression of cell cycle, inhibit the proliferation of HSC, inhibit NF - ”B related signaling pathway, promote the apoptosis of HSC, regulate the synthesis and decomposition of cytoskeleton and extracellular matrix, Reduce hepatic collagen deposition.Conclusion: I3C can exert its anti-hepatic fibrosis through multiple pathways.