叶下珠复方Ⅱ号对肝星状细胞TGF-β1及Ⅰ型胶原的影响

Impact of compound phyllanthus urinaria(CPU Ⅱ)on the expression of TGF-β1 and collagen Ⅰ synthesis in hepatic stellate cells

目的观察叶下珠复方Ⅱ号体外对肝星状细胞(HSC-T6)TGF-β1mRNA、Ⅰ型胶原mRNA表达及其蛋白合成的影响,进一步探讨叶下珠复方Ⅱ号抗肝纤维化的作用机制。方法体外培养HSC-T6细胞,分为对照组、叶下珠复方Ⅱ号高剂量组(3.60mg/ml)、中剂量组(1.80mg/ml)和低剂量组(0.9mg/ml),采用MTT法检测药物对HSC-T6细胞增殖的影响,SYBR染料法实时荧光定量PCR检测药物处理后HSC-T6细胞TGF-β1及Ⅰ型胶原mRNA的表达变化,Western Blot法检测I型胶原蛋白表达的改变。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、TGF-β1mRNA及Ⅰ型胶原合成呈现显著的抑制作用,与细胞对照组比较,差异均有统计学意义(P<0.05和0.01),而且随着药物浓度的增加其抑制作用明显加强。结论叶下珠复方Ⅱ号能显著抑制HSC-T6细胞增殖、明显降低TGF-β1mRNA、Ⅰ型胶原mRNA的表达及其蛋白合成,具有明显的抗肝纤维化作用。

Objective To observe the impact of compound phyllanthus urinarian（CPUⅡ）on the expression of TGF-β1and collagen Ⅰ synthesis in hepatic stellate cells,and explore the antifibrotic mechanisms of CPUⅡ.Methods HSC-T6 cells were cultured in vitro and divided into 4 groups,including control group,CPUⅡ 3.60 mg/ml group,CPUⅡ 1.8mg/ml group,CPUⅡ 0.90mg/ml group.The level of TGF-β1 and collagen I mRNA were analyzed by real time PCR;The level of collagen Ⅰ proteins were analyzed by Western blotting.Results After CPUⅡ impacted on HSC-T6,real time PCR results showed that the leve of TGF-β1 and collagen Ⅰ mRNA in every treatment group of CPUⅡwere significantly lower than that of control group（P〈0.05、0.01）.Westen blotting results showed that the expression of collagen Ⅰ was inhibited significantly.Conclusion CPUⅡ is capable of regulating the expression of TGF-β1 and collagen Ⅰ synthesis in hepatic stellate cells and have the capability of antifibrosis.