葛根素和小檗碱联合用药干预NASH细胞模型的实验研究

• 作 者: 王允亮 叶杨 程佳伟 高康丽 赵唯含 刘丽娟 李军祥
• 机 构: 北京中医药大学东方医院消化内科,
• 出 处: 中国中西医结合消化杂志
• 发表日期: 2015年05期
• 关键词: NASH 细胞模型 葛根素 小檗碱 联合用药 空白对照组 模型组 葛根素组 检测细胞培养 油红

[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 explore the related dosages and effects of puerarin and berberine in the intervention of non-alcoholic steatohepatitis model. [Methods] HepG2 cells were induced by 500μmol / L FFA for 24 hours, and NASH cell model was established. NASH cells were treated with different concentrations of puerarin and berberine alone and in combination. MTT assay was used to detect the cell viability Safe drug concentration range; oil red O staining intracellular fatty degeneration; Elisa method detection of cell culture supernatant TNF-α, IL-8 and GLUT4 levels. [Results] The content of TNF-α and IL-8 in 1NASH cell model group was significantly higher than that of blank control group, while the content of GLUT4 was significantly lower than that of blank control group. 2 Different concentrations of puerarin and berberine alone and combination therapy group , With the increase of berberine and puerarin alone, the levels of IL-8 and TNF-a were significantly lower than those of the model group, the content of GLUT4 was significantly increased and the number of red lipid droplets was decreased; , TNF-a, GLUT4 levels and intracellular red lipid droplets were significantly improved compared with single drug treatment. [Conclusion] The combination therapy of puerarin and berberine has synergistic effect in improving the cytopathology, the levels of TNF-α, IL-8 and GLUT4, and the combined effect is better than the single drug. Cell experiments, the basic can be determined within a certain range, berberine and puerarin combination ratio of 1  40 ~ 1  10 is better.