人参皂苷Rd抑制肝星状细胞活化作用与机制研究

Ginsenoside Rd attenuates the activation of hepatic stellate cells induced by oxidized low-density lipoprotein

目的:观察人参皂苷Rd对氧化低密度脂蛋白(OX-LDL)诱导肝星状细胞(HSCs)活化与分泌胶原作用的影响及其机制。方法:10μg/ml OX-LDL孵育诱导肝星状细胞活化建立体外肝纤维化模型,分别给予人参皂苷Rd(18.8mg/L或37.6mg/L)处理肝星状细胞48小时。MTT法检测肝星状细胞增殖,荧光定量PCR检测转化生长因子(TGF)-β1和CD36 m RNA表达;Western blot检测I型胶原及CD36、Smad2、Smad4表达。结果:与正常对照组相比,模型处理组的I型胶原蛋白表达与TGF-β1 m RNA表达均明显增加;与模型处理组相比,人参皂苷Rd(38.7mg/L)明显抑制肝星状细胞增殖,降低CD36、I型胶原和Smad4蛋白表达,同时,人参皂苷Rd对Smad2蛋白表达没有影响。结论:人参皂苷Rd可能通过降低CD36及Smad4蛋白表达减少细胞外胶原蛋白的合成,进而发挥抗纤维化作用。

Objective： To observe the effects of ginsenoside Rd on the activation of hepatic stellate cells （HSCs） induced by OX-LDL in vitro. Methods： HSCs were cultured in DMED in vitro. Ten microgram of oxidized low-density lipoprotein （OX-LDL） was used to induce the acti- vation of HSCs. And HSCs were incubated with 18.8mg/L or 37.6mg/L of ginsenoside Rd for 48 hours, respectively. Proliferation of HSCs was determined by MTT assay, mRNA expression levels of transforming growth factor （TGF）-β1 and CD36 were determined by real-time PCR technique. And protein expression of collagen type I, CD36, Smad2 and Smad4 were measured by Western blot. Results： The protein expression of collagen I and TGF-β1 mRNA significantly increased in HSCs stimulated with OX-LDL by comparison of those in the control group, respectively. Ginsenoside Rd （37.6mg/L） markedly inhibited proliferation of HSCs, decreased protein expressions of CD36, collagen I and Smad4 （P 〈 0.01 ）. Meanwhile, it displayed no obvious effect on Smad2 protein expression. Conclusion： Ginsenoside Rd inhibits col- lagen type I protein expression via down-regulating CD36 expression, and decreases the synthesis of Smad4, which exerts a protective effect on liver fibrosis.