BZL方对游离脂肪酸诱导HepG2细胞脂肪沉积和LKB1-AMPK-ACC信号传导通路的影响

Effects of BZL Decoction on fatty deposition in HepG2 cells induced by free fatty acid and on LKB1-AMPK-ACC signal pathway

目的:研究"祛湿化瘀方"中3种已被认知的、能被分离提取的有效组分(白术多糖、栀子苷、绿原酸)组成的BZL复方对体外肝脂毒性模型脂肪沉积和LKB1-AMPK-ACC信号传导通路的影响,以进一步探讨BZL方防治脂肪肝的作用机制。方法:建立游离脂肪酸诱导的HepG2细胞脂肪变性模型,设正常组、模型组和不同浓度药物血清组分别观察细胞上清液中的细胞内甘油三酯(TG)含量,细胞油红O染色,细胞的P-LKB1、P-AMPKα、P-ACC蛋白表达及LKB1、AMPKα、ACC基因表达。结果:FFA刺激24h后,模型组细胞内脂肪沉积明显,其TG含量明显高于正常组(P<0.01),细胞内P-LKB1、P-AMPK、P-ACC的蛋白表达均减弱,LKB1、AMPK、ACC的mRNA表达均明显减弱,而BZL方高剂量组细胞内TG含量显著低于模型组(P<0.01),其脂肪沉积与模型组相比也明显减轻;与模型组比较,BZL药物血清组细胞内P-LKB1、P-AMPKα、P-ACC的蛋白表达均明显增强(P<0.05,P<0.01),LKB1、AMPKα、ACC的mRNA表达也均明显增强(P<0.05,P<0.01)。结论:BZL方对FFA诱导的HepG2细胞脂肪沉积有显著的抑制作用,且呈剂量依赖关系,激活LKB1-AMPK-ACC信号传导通路可能是BZL方防治非酒精性脂肪性肝病的重要作用机制之一。

Objective： To investigate the effects of BZL Decoction on fatty deposition and on LKB1-AMPK-ACC signal pathway of hepatic lipotoxicity model in vitro, and to explore the mechanism of BZL Decoction on preventing and treating nonalcoholic fatty liver disease （NAFLD）. Methods： The steatosis modeling of HepG2 cells were induced by free fatty acid （FFA）. HepG2 cells were divided into a control group, a model group and treatment groups. Treatment groups were administrated with different concentrations of BZL Decoction contained serum. The contents of TG in cells were measured. The pathological changes of cells were observe by oil-red O staining. The P-LKBI, P-AMPK, P-ACC protein expression and LKB1, AMPKa, ACC expression were measured respectively. Results： After stimulated with FFA for 24 hours, model group showed remarkable deposition of lipid. Compared with the control group, the content of TG in model group was much higher （P〈0.01）. All of the protein expression of P-LKBI, P-AMPK and P-ACC significantly decreased, and all of mRNA expression of LKB1, AMPK and ACC significantly decreased. Compared with the content of TG in the model group, the content of TG in high-doze BZL Decoction group was much lower （P〈0.01）, and the deposition of lipid was much lower. Compared with the model group, all of the protein expression of P-LKB1, P-AMPK and P-ACC in treatment groups significantly increased, and all of mRNA expression of LKB1, AMPK and ACC significantly increased （P〈0.05, P〈0.01）. Conclusion： BZL Decoction can significantly inhibit thefatty deposition in HepG2 cells induced by FFA. The effect is in dose-dependent manner. To activate LKB 1-AMPK-ACC signal pathway might be one of the most important mechanisms of BZL Decoction＇s prevention and treatment of NAFLD.