摘要
目的在细胞模型中研究白芍总苷抗非酒精脂肪肝的作用机制。方法采用油酸干预HepG2细胞构建脂肪肝细胞模型,油红O染色观察细胞内脂肪滴。采用CCK-8检测白芍总苷对细胞增殖能力的影响,并筛选出最佳的作用浓度和作用时间。将细胞分为4组:对照组、模型组、白芍总苷预处理组和治疗组。生化实验检测细胞总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、丙二醛(malondialdehyde,MDA)含量及碱性磷酸酶(alkaline phosphatase,ALP)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSHPx)、丙氨酸氨基转移酶(alanine aminotransferase,ALT)活性。透射电子显微镜观察细胞线粒体的超微结构。Real-time PCR检测细胞酰基辅酶A氧化酶1(acyl coenzyme A oxidase1,ACOX1)基因表达水平。Western blot检测细胞线粒体融合蛋白1(Mitofusin 1,Mfn1)、线粒体融合蛋白2(Mitofusin 2,Mfn2)、视神经萎缩蛋白1(Optic atrophy 1,Opa1)和发动蛋白相关蛋白1(Dynamin-related protein 1,Drp1)表达水平。结果成功构建脂肪肝细胞模型,筛选出白芍总苷的最佳的作用浓度为80μg/mL,作用时间为24 h。与对照组比较,模型组TC、TG、MDA浓度及ALP和ALT活性均显著升高(P<0.05),GSH-Px活性显著降低(P<0.05),线粒体肿胀,嵴稀疏、断裂、溶解,逐渐空泡化,ACOX1 mRNA和Mfn1、Mfn2、Opa1蛋白表达水平显著降低(P<0.05),Drp1蛋白表达水平显著升高(P<0.05)。与模型组比较,白芍总苷预处理组和治疗组细胞TC、TG、MDA浓度及ALP和ALT活性均显著降低(P<0.05),GSH-Px活性显著升高(P<0.05),大部分线粒体膜结构较为完整,嵴结构清晰且紧密排列,ACOX1 mRNA和Mfn1、Mfn2、Opa1蛋白表达水平显著升高(P<0.05),Drp1蛋白表达水平显著降低(P<0.05)。结论白芍总苷对脂肪肝具有较好的防治作用,其作用机制与改善线粒体损伤,保护线粒体功能有关。
Objective To study the anti-fatty liver action mechanisms of total glucosides of paeony in a fatty liver cell model.Methods HepG2 cells were intervened with oleic acid to establish the model of fatty liver cells.The lipid droplets were observed by oil red O staining.CCK-8 was used to detect the effect of total glucoside of peony on cell proliferation ability and to screen the optimal concentration and duration of action.The cells were divided into four groups:control,model,total glucosides of paeony pretreatment and treatment.The total cholesterol(TC),triglyceride(TG),malondialdehyde(MDA)and the activities of alkaline phosphatase(ALP),glutathione peroxidase(GSH-Px),alanine aminotransferase(ALT)were detected by biochemical experiments.The mitochondrial ultrastructure of cells was observed by transmission electron microscopy.The gene expression of acyl coenzyme A oxidase 1(ACOX1)was detected by real-time PCR.The expression levels of mitofusin 1(mfn1),mitofusin 2(Mfn2),optical atrophy 1(OPA1)and dynamic related protein 1(drp1)were detected by western blot.Results Successfully constructing fatty liver cell model.The optimal concentration of total glycosides of peony was screened as 80μg/mL and the duration of action was 24 h.Compared with the control group,the concentrations of TC,TG,MDA and the activities of ALP and ALT were significantly increased(P<0.05),while the activity of GSH-Px was significantly decreased(P<0.05).Mitochondrial swelling,sparse cristae,fracture,dissolution,gradually vacuolated,and the expression of ACOX1 mRNA and the protein expression of Mfn1,Mfn2 and Opa1 were significantly decreased(P<0.05),and Drp1 protein expression level was significantly increased in the model group(P<0.05).Compared with the model group,the concentrations of TC,TG,MDA and the activities of ALP and ALT were significantly decreased(P<0.05),while the activity of GSH-Px was significantly increased(P<0.05).Most of the mitochondrial membrane structure was relatively complete,the cristae structure was clear and closely arranged,and the expression of ACOX1 mRNA and the protein expression of Mfn1,Mfn2,and Opa1were significantly increased(P<0.05),and Drp1 protein expression level was significantly decreased in the total glucosides of paeony pretreatment and treatment groups(P<0.05).Conclusion Total glucosides of paeony has a good preventive and therapeutic effect on fatty liver cells,and its mechanism is related to improving mitochondrial damage and protecting mitochondrial function.
作者
孙智珺
叶阳梅
王光新
Sun Zhijun;Ye Yangmei;Wang Guangxin(Wuhan Third Hospital,Wuhan 430000,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2022年第7期2748-2754,共7页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
关键词
脂肪肝
白芍总苷
线粒体结构
线粒体功能
Fatty liver
Total glucosides of paeony
Mitochondrial structure
Mitochondrial function
作者简介
通讯作者:王光新,副主任医师,硕士,主要研究方向:普通外科。
