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目的:观察三甲散含药血清对HSC-T6细胞增殖及对Rho/ROCK信号通路的影响。方法:以三甲散含药血清作用HSC-T6细胞,以MTT法检测其对HSC-T6细胞增殖的影响,ELISA检测其对Ⅰ、Ⅲ型胶原合成的影响,荧光定量PCR检测其对α-SMA、TIMP-1、ROCK-ⅠmRNA表达的影响,Western blot检测其对α-SMA、TIMP-1、ROCK-Ⅰ蛋白表达的影响。结果:三甲散含药血清对HSC-T6细胞增殖抑制率从12 h开始升高,24 h达到峰值;三甲散组细胞Ⅰ、Ⅲ型胶原含量较空白对照组显著降低(P<0.05或P<0.01);三甲散和Y-27632均能下调HSC-T6细胞α-SMA、ROCK-Ⅰ、TIMP-1 mRNA及蛋白的表达(P<0.05或P<0.01)。结论:三甲散可抑制HSC-T6细胞的增殖,其机制可能与Rho/ROCK信号通路有关。
OBJECTIVE: To observe the effects of SJA serum on the proliferation of HSC-T6 cells and the effect on Rho / ROCK signaling pathway. Methods: The HSC-T6 cells were treated with Sanjian San containing serum and the effects of SJT-HSC-T6 cells on the proliferation of HSC-T6 cells were detected by MTT assay. The effects on the synthesis of type I and type III collagen by ELISA were detected by fluorescence quantitative PCR. , TIMP-1, ROCK-ⅠmRNA expression, Western blot was used to detect the expression ofα-SMA, TIMP-1, ROCK-Ⅰprotein. Results: The inhibitory rate of SJA-containing serum on HSC-T6 cells increased from 12 h to 24 h, and reached the peak at 24 h. The contents of type I and type III collagen in SJG cells were significantly lower than those in control group (P <0.05 or P < 0.01) .SMA and Y-27632 both down-regulated the mRNA and protein expressions of α-SMA, ROCK-Ⅰ and TIMP-1 in HSC-T6 cells (P <0.05 or P <0.01). Conclusion: Sanjian Powder can inhibit the proliferation of HSC-T6 cells, which may be related to the Rho / ROCK signaling pathway.