柔肝方含药血清对TGF-β1诱导人肝星状细胞表达骨桥蛋白的影响及其机制  

Effects and Mechanisms of Serum Containing Rougan Decoction on Expression of Osteopontin in Human Hepatic Stellate Cells Induced by TGF-β_1

作　　者：吴惠春[1] 张斌[1] 王灵台[1] 

机构地区：[1]上海中医药大学附属曙光医院,上海201203

出　　处：《中国实验方剂学杂志》2015年第13期109-113,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：上海市教育委员会重点学科建设项目(J50307)

摘　　要：目的:研究柔肝方含药血清对转化生长因子β1(TGF-β1)诱导人肝星状细胞表达骨桥蛋白(osteopontin,OPN)的影响及其作用机制。方法:30只SD大鼠,随机分成正常组、柔肝方低剂量(9 g·kg-1)给药3 d及5 d组、柔肝方高剂量(18 g·kg-1)给药3 d及5 d组,每组6只,按10 m L·kg-1·d-1灌服饮用水或柔肝方药液,制备含药血清。用含10%正常血清或含药血清的DMEM培养基体外培养LX-2人肝星状细胞株,MTT法检测细胞增殖;LX-2细胞经含10%正常血清或含药血清的培养基预处理24 h,再予TGF-β1(终质量浓度10μg·L-1)刺激24 h,收集细胞,RT-PCR法检测OPN mRNA表达,Western blot法检测OPN及磷酸化蛋白激酶B(p-protein kinase B,p-PKB)蛋白表达。结果:与正常组比较,各含药血清组均能抑制LX-2细胞增殖(P<0.05,P<0.01);予含药血清预处理再经TGF-β1刺激的细胞,与正常组细胞比较,OPN mRNA,OPN和p-PKB蛋白表达明显下调(P<0.05,P<0.01)。结论:柔肝方含药血清对TGF-β1诱导的人肝星状细胞表达OPN具有抑制作用,其机制可能与抑制PI3K/PKB信号通路活化有关。Objective： To observe the effects and mechanisms of serum containing Rougan decoction on expression of osteopontin in human hepatic stellate cells induced by TGF-β1. Method： Thirty rats were randomly divided into 5 groups： the control group,the 3 /5 d groups with low-dose Rougan decoction（ 9 g·kg- 1）,the 3 /5d groups with high-dose Rougan decoction（ 18 g·kg- 1） of 6 rats each. Rats in each group received distilled water or Rougan decoction at 10 m L·kg- 1·d- 1and the corresponding serums containing drug were obtained. Human hepatic stellate cell line LX-2 was cultured in DMEM containing 10% control serum or serum containing Rougan decoction. The proliferation of LX-2 cells was detected by using MTT method. LX-2 cells were pretreated with control serum or serum containing Rougan decoction for 24 h followed by incubation with TGF-β1at the final concentration of 10 μg·L- 1for 24 h. The expression of OPN was detected by real-time PCR and Western blot,the expression of p-PKB was detected by Western blot. Result： Compared with the control group,the proliferation of LX-2 cells in serum containing Rougan decoction groups was significantly inhibited（ P〈0. 05,P〈0. 01）.Meanwhile,the expression of OPN and p-PKB were significantly down-regulated in serum containing Rougan decoction groups（ P〈0. 05,P〈0. 01）. Conclusion： The serum containing Rougan decoction has an inhibitory effect on the expression of OPN induced by TGF-β1,the mechanism may be related to the inhibition of PI3K/PKB signaling pathway.

关 键 词：柔肝方 骨桥蛋白 肝星状细胞 转化生长因子β1 

分 类 号：R285.5[医药卫生—中药学]