葛根芩连汤含药血清对游离脂肪酸诱导的HepG2细胞脂肪堆积模型FXR通路相关信号因子的影响

Effects of Gegen Qinlian Decoction containing serum on FXR pathway related signal factors in free fatty acid-induced adipose accumulation model of HepG2 cells

目的利用体外游离脂肪酸(Free Fat Acid,FFA)诱导的HepG2细胞脂肪堆积模型,分析葛根芩连汤含药血清对FXR通路相关信号因子的影响。方法用1mmol/L FFA诱导HepG2细胞建立脂肪堆积模型,给予不同浓度的葛根芩连汤和吡格列酮含药血清进行干预。采用TG试剂盒法检测HepG2细胞脂肪变性状况。WB法检测HepG2细胞内FXR蛋白表达水平。qRT-PCR法检测各组HepG2细胞内FXR、CYP7A1、SHP、FGFR4和TGR5 mRNA表达程度。结果葛根芩连汤中剂量组、低剂量组的TG含量比模型组显著降低(P<0.01,P<0.05)。WB实验表明,与模型组相比,葛根芩连汤中剂量组与吡格列酮组FXR蛋白表达量显著增加(P<0.01,P<0.05)。qRT-PCR实验表明,与模型组相比,葛根芩连汤高、中、低剂量组FXR、SHP、FGFR4和TGR5 mRNA表达水平均显著升高(P<0.01),CYP7A1 mRNA表达水平均显著降低(P<0.01,P<0.05)。吡格列酮组内FXR、SHP、FGFR4和TGR5 mRNA表达水平显著升高(P<0.01,P<0.05),CYP7A1 mRNA表达水平显著降低(P<0.01)。结论葛根芩连汤能够减轻FFA诱导的HepG2细胞脂肪变性,其机制可能与FXR信号通路有关。

Objective To investigate the effects of Gegen Qinlian Decoction containing serum on FXR pathway related signal factors in fat accumulation model of HepG2 cells induced by free fat acid(FFA) were analyzed by using fat accumulation model of HepG2 cells in vitro.Methods HepG2 cells were induced by 1 mmol/L FFA to establish fat accumulation model, and treated with different concentrations of Gegen Qinlian Decoction and pioglitazone containing serum. The steatosis of HepG2 cells was detected by TG kit method. The expression level of FXR protein in HepG2 cells was detected by western blot method. The mRNA expression levels of FXR, CYP7 A1, SHP, FGFR4 and TGR5 in HepG2 cells were detected by qRT-PCR method.Results The TG content in Gegen Qinlian Decoction medium-dose and low-dose groups were significantly lower than that in model group(P<0.01, P<0.05). Compared with model group, results of the western blot experiment showed the expression of FXR protein in Gegen Qinlian Decoction medium-dose group and pioglitazone group were significantly increased(P<0.01, P<0.05). Compared with model group, the results of qRT-PCR experiment showed the mRNA expression levels of FXR, SHP, FGFR4 and TGR5 in Gegen Qinlian Decoction high-dose, medium-dose and low-dose groups were significantly increased(P<0.01), CYP7 A1 mRNA expression levels were significantly decreased(P<0.01, P<0.05);the mRNA expression levels of FXR, SHP, FGFR4 and TGR5 in pioglitazone group were significantly increased(P<0.01, P<0.05), CYP7 A1 mRNA expression level was significantly decreased(P<0.01).Conclusion Gegen Qinlian Decoction can reduce FFA-induced steatosis of HepG2 cells, and the mechanism may be related to FXR signaling pathway.