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目的:探讨抗纤软肝颗粒含药血清对肝星状细胞黏着斑激酶(FAK)mRNA、蛋白表达的影响。方法:运用细胞培养技术,以hsc-t6细胞系为研究对象,分为(1)空白包被+10%生理盐水血清组;(2)纤维连接蛋白(FN)+10%生理盐水血清组;(3)FN+5%抗纤软肝颗粒含药血清(kxr)组;(4)FN+10%kxr组;(5)FN+20%kxr组。按以上分组进行药物干预,干预后继续培养48 h。应用免疫细胞组织化学方法检测FAK蛋白的表达,采用RT-PCR法测定FAK mRNA的表达。结果:kxr组肝星状细胞的FAK mRNA、蛋白表达均显著下调,分别减少了72.78%和27.88%,78.82%和44.85%,85.51%和57.58%,与对照组及FN组相比,5%kxr组(P<0.05),10%kxr组、20%kxr组(P<0.01)。结论:kxr能在转录及翻译水平抑制FAK mRNA、蛋白的表达,从而阻抑了整合素激活的FAK途径的信号转导。
Objective: To investigate the effect of anti-fibrosis liver granule drug-containing serum on the expression of focal adhesion kinase (FAK) mRNA and protein in hepatic stellate cells. METHODS: Using cell culture techniques, the hsc-t6 cell line was used as the study object and divided into (1) blank-coated +10% saline sera and (2) fibronectin (FN) + 10% saline sera. (3) FN+5% anti-fibrosis liver granule drug-containing serum (kxr) group; (4) FN+10% kxr group; (5) FN+20% kxr group. According to the above groups for drug intervention, the intervention continued after 48 h. The expression of FAK protein was detected by immunohistochemistry and the expression of FAK mRNA was determined by RT-PCR. RESULTS: FAK mRNA and protein expression in hepatic stellate cells were significantly down-regulated in the kxr group, which were reduced by 72.78% and 27.88%, 78.82% and 44.85%, 85.51% and 57.58%, respectively, compared with the control and FN groups, 5% Kxr group (P <0.05), 10% kxr group, 20% kxr group (P <0.01). Conclusion: Kxr can inhibit the expression of FAK mRNA and protein at the transcriptional and translational levels, thus inhibiting the signal transduction of integrin-activated FAK pathway.