胡黄连中环烯醚萜苷类成分体外抗非酒精性脂肪性肝炎活性研究

Anti-NASH activity of Iridoid Glycosides in Picrorhizae Rhizoma in vitro

目的探究胡黄连苷Ⅰ、Ⅱ、Ⅲ、Ⅳ体外抗非酒精性脂肪性肝炎(NASH)的活性。方法用1 mmol/L游离脂肪酸(FFA)诱导HepG2细胞24 h,建立体外NASH模型;MTT法检测胡黄连苷Ⅰ、Ⅱ、Ⅲ、Ⅳ对细胞活力的影响,并依此优选3个安全给药浓度(0.25、0.50、1.00 mg/mL)干预体外NASH模型;油红O染色法镜下观察细胞内脂滴的变化情况;试剂盒法检测细胞培养上清液中超氧化物歧化酶(SOD)、丙二醛(MDA)、白介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)的含量变化。结果胡黄连苷Ⅰ、Ⅱ、Ⅲ、Ⅳ对HepG2细胞的活力影响较小。FFA刺激24 h后,模型组与对照组比较,细胞内被染成明显的红色颗粒状脂滴,细胞培养上清液中SOD水平显著下降,MDA、TNF-α、IL-8水平显著上升,均有显著性差异(P<0.05、0.01)。与模型组比较,不同浓度的胡黄连苷Ⅰ、Ⅱ、Ⅲ、Ⅳ干预组,细胞内红色颗粒状脂滴减少,且以胡黄连苷Ⅱ最为明显;SOD水平显著上升,MDA、TNF-α、IL-8水平显著下降(P<0.05、0.01),且以胡黄连苷Ⅱ、Ⅲ、Ⅳ较为明显,但没有显著的剂量相关性。结论胡黄连苷Ⅰ、Ⅱ、Ⅲ、Ⅳ中胡黄连苷Ⅱ的体外抗NASH的活性较显著。

Objective To discuss the anti-NASH activity of PicrosideⅠ,PicrosideⅡ,PicrosideⅢand PicrosideⅣin vitro.Methods Liver cells HepG2 were cultured and induced by FFA to establish NASH cell model.The cell viability were measured by MTT to select the safe drug concentration.Then safe concentrations of PicrosideⅠ,PicrosideⅡ,PicrosideⅢand PicrosideⅣwere meanwhile added.The fat deposition in cells was observed by oil-red O staining method.The content of SOD,MDA,IL-8 and TNF-αwere tested by Elisa.Results Three treatment groups were seted with the concentration of 0.25,0.50 and 1.00 mg/mL.Compared with control group,the NASH cell model group was stained with obvious red granular lipid droplets.The levels of SOD were significantly higher in model group than the control group,while MDA,TNF-αand IL-8 significantly lower than control group(P<0.05 and 0.01).Compared with the model group,the red granular lipid droplets decreased in the intervention groups of different concentrations of PicrosideⅠ,ⅡⅢandⅣ,especially Picroside II.SOD levels increased significantly,MDA,TNF-a and IL-8 levels decreased significantly(P<0.05,0.01),and PicrosideⅡⅢandⅣwere more obvious,but there was no significant dose correlation.Conclusion PicrosideⅡcan improve cellular pathology and influence the levels of SOD,MDA,IL-8 and TNF-αobviously,showing potential value on NASH treatment.