葛根素和小檗碱联合用药干预NASH细胞模型的实验研究

Experimental studies of the combination effects of berberine and puerarin on NASH in vitro

[目的]探讨葛根素、小檗碱联合用药干预非酒精性脂肪性肝炎细胞模型的相关剂量及作用效果。[方法]用500μmol/L的FFA诱导HepG2细胞24h,建立NASH细胞模型;用不同浓度的葛根素、小檗碱单药及组合用药干预NASH细胞模型,MTT法检测两种单体对细胞模型的安全药物浓度范围;油红O染色观察细胞内脂肪变;Elisa法检测细胞培养上清液中TNF-α、IL-8及GLUT4水平。[结果]1NASH细胞模型组培养液中TNF-α及IL-8含量明显高于空白对照组,GLUT4含量明显低于空白对照组;2不同浓度葛根素、小檗碱单用及联合用药干预组,随着小檗碱、葛根素单独使用浓度的增加,IL-8、TNF-a水平较模型组明显降低,GLUT4含量显著增加,细胞内红色脂滴减少;单体联合用药后,IL-8、TNF-a、GLUT4水平及细胞内红色脂滴均较单独用药改善明显。[结论]葛根素、小檗碱组合用药在改善细胞病理、TNF-α、IL-8、GLUT4水平方面存在协同作用,其组合用药效果较单独用药效果增强。细胞实验中,基本能够确定在一定范围内,小檗碱与葛根素组合比例在140-110为佳。

[Objective]To discuss the combination effects of puerarin and berberine on NASHin vitro.[Methods]Liver cells HepG2 were cultured and induced by FFA to establish NASH model.Then different concentrations of puerarin and berberine combination with different proportions were meanwhile added.MTT was used to detect IL-8,TNF-α,and GLUT4 in supernatant.Oil red O staining was used to observe the adipose cells.[Results]（1）The levels of TNF-αand IL-8were significantly higher in model group than the control group,while GLUT4 significantly lower than control group.（2）Different concentrations of puerarin and berberine alone and combination in the intervention groups decreased lipid droplets significantly and increased GLUT4 level in a dose-dependent manner,with better results in the combination group than single usage of puerarin and berberine groups.[Conclusion]Combination of puerarin and berberine can improve cellular pathology and influence the levels of TNF-α,IL-8and GLUT4,showing a synergistic effect.