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目的观察姜黄素干预前后肝星状细胞(HSC)中髓样分化因子88(My D88)蛋白的变化水平,以探讨姜黄素抗肝纤维化的相关机制。方法构建p CMV-Myc-My D88重组质粒,将人肾上皮细胞293T细胞及HSC-T6细胞分别设为空白对照组、My D88过表达组、姜黄素+My D88过表达组,其中My D88过表达组给予转染p CMV-Myc-My D88重组质粒72 h,姜黄素+My D88过表达组在转染48 h后加入姜黄素继续作用24 h。采用Western blot法检测My D88蛋白表达。结果 1转染p CMV-mycMy D88重组质粒48 h后,My D88在293T细胞及HSC中表达均明显增高(P<0.05),且经姜黄素处理后两类细胞中表达量明显降低(P<0.05)。2姜黄素处理后My D88表达量较My D88质粒转染组明显降低(P<0.05)。结论姜黄素可能通过抑制肝星状细胞My D88蛋白表达,阻断My D88依赖性信号通路的转导而发挥抗肝纤维化的作用。
Objective To observe the changes of myeloid differentiation factor 88 (My D88) protein in hepatic stellate cells (HSC) before and after the intervention of curcumin so as to explore the mechanism of curcumin in preventing hepatic fibrosis. Methods The recombinant plasmid pCMV-Myc-My D88 was constructed and the human renal epithelial 293T cells and HSC-T6 cells were set as blank control group, My D88 overexpression group, Curcumin + My D88 overexpression group, The recombinant plasmid was transfected with pCMV-Myc-My D88 plasmid for 72 h. The curcumin + My D88 overexpression group was treated with curcumin 48 h after transfection for 24 h. Western blot was used to detect My D88 protein expression. Results After transfected with recombinant plasmid p CMV-mycMy D88 for 48 h, the expression of My D88 in 293T cells and HSCs was significantly increased (P <0.05), and the expression of My D88 in both groups decreased significantly after treatment with curcumin (P < 0.05). 2 Curcumin treatment My D88 expression was significantly lower than the My D88 plasmid transfected group (P <0.05). Conclusion Curcumin may play an anti-hepatic fibrosis role by inhibiting My D88 protein expression in the hepatic stellate cells and blocking the transduction of My D88-dependent signaling pathway.