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【目的】观察叶下珠复方Ⅱ号体外对肝星状细胞(HSC-T6)凋亡和miR-16/Bcl-2表达的影响,进一步探讨其抗肝纤维化的作用机制。【方法】体外培养HSC-T6细胞,分为空白对照组,叶下珠复方Ⅱ号高剂量组(3.6 g/L)、中剂量组(1.8 g/L)和低剂量组(0.9 g/L),采用四甲基偶氮唑盐(MTT)法检测药物对HSC-T6细胞增殖的影响,流式细胞仪检测细胞凋亡的变化,实时荧光定量PCR法检测药物处理后HSC-T6细胞miR-16和Bcl-2 mRNA表达的改变,Western Blot法测定Bcl-2蛋白的表达变化。【结果】叶下珠复方Ⅱ号处理HSC-T6细胞后,高、中、低各剂量组对HSC-T6细胞增殖均有明显的抑制作用,随着叶下珠复方Ⅱ号浓度的增加,细胞的凋亡率也逐渐增加;叶下珠复方Ⅱ号各处理组miR-16的表达显著增加,呈现明显的剂量—效应关系,且Bcl-2 mRNA和蛋白质表达显著减少,与空白对照组比较,差异均具有统计学意义(P<0.01)。【结论】叶下珠复方Ⅱ号能显著抑制HSC-T6细胞增殖和诱导其凋亡,其作用机制与提高miR-16的表达,抑制其靶基因Bcl-2的表达有关。
【Objective】 To observe the effect of Phyllanthus Compound Ⅱ on the apoptosis of HSC-T6 and the expression of miR-16 / Bcl-2 in vitro, and to explore its mechanism of action against hepatic fibrosis. 【Methods】 HSC-T6 cells were cultured in vitro and divided into three groups: blank control group, Phytophyllum compound II high dose group (3.6 g / L), middle dose group (1.8 g / L) and low dose group ), The effects of drugs on the proliferation of HSC-T6 cells were detected by MTT assay, the apoptosis of HSC-T6 cells was detected by flow cytometry, and the expression of miRs in HSC-T6 cells was detected by real-time fluorescence quantitative PCR -16 and Bcl-2 mRNA expression changes, Western Blot determination of Bcl-2 protein expression changes. 【Results】 The results showed that Phytophyllum compound II could inhibit the proliferation of HSC-T6 cells in high, medium and low dose groups. With the increase of the concentration of Phyllanthus Compound II, And the apoptotic rate increased gradually. The expression of miR-16 in each treatment group was significantly increased, the dose-effect relationship was obvious, and the expression of Bcl-2 mRNA and protein was significantly decreased. Compared with the blank control group, The differences were statistically significant (P <0.01). 【Conclusion】 Ye Xia Zhu Fu Fang Ⅱ significantly inhibits the proliferation and induces apoptosis of HSC-T6 cells, and its mechanism may be related to increasing the expression of miR-16 and inhibiting the expression of its target gene Bcl-2.