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目的:观察氧化苦参碱对高脂诱导的HepG2细胞脂质沉积的干预作用及其机制。方法:高脂干预法培养HepG2细胞建立非酒精性脂肪性肝(nonalcoholic fatty liver disease,NAFLD)体外细胞模型。造模成功后随机分为高脂组(Pa组)、氧化苦参碱组(Omt组)和二甲双胍组(Met组),同时设立正常培养基对照组(N组)。药物干预48 h后收集细胞,利用相应试剂盒及2,7-二氯氢化荧光素二乙酸酯(2,7-dichlorofuorescin diacetate,DCFH-DA)荧光探针评估各组细胞内的脂质沉积水平和氧化应激状态;通过PCR和Western blot分别检测参与脂代谢关键分子的基因和蛋白表达。结果:与Pa组相比,氧化苦参碱及二甲双胍干预后可显著降低细胞内甘油三酯(triglyceride,TG)含量(POmt=0.023,PMet=0.043),上调肉碱脂酰转移酶-1(carnitine palmitoyl transferase-1,CPT-1)的m RNA表达(POmt=0.000,PMet=0.000),下调脂肪酸转位酶(fatty acid translocase,FAT/CD36)的m RNA(POmt=0.000,PMet=0.000)及蛋白表达水平(POmt=0.000,PMet=0.000);DCFH-DA荧光染色、总超氧化物歧化酶(total superoxide dismutase,T-SOD)、谷胱甘肽过氧化物酶(glutathion peroxidase,GSH-Px)及丙二醛(malondialdehyde,MDA)的测定结果显示药物干预可显著增强T-SOD(POmt=0.007,PMet=0.007)及GSH-Px(POmt=0.183,PMet=0.020)的活性,减少活性氧(reactive oxygen species,ROS)和MDA(POmt=0.016,PMet=0.001)的生成。结论:氧化苦参碱可改善高脂诱导的HepG2细胞细胞内脂质沉积,可能是通过改善脂代谢及细胞内氧化应激状态实现的。
Objective: To observe the intervention of oxymatrine on lipid-induced lipid deposition in HepG2 cells and its mechanism. Methods: HepG2 cells were cultured with high fat diet to establish an in vitro cell model of nonalcoholic fatty liver disease (NAFLD). The rats were randomly divided into high fat group (Pa group), oxytocin group (Omt group) and Metformin group (Met group). Normal control group (N group) was also established. After 48 h of drug intervention, the cells were harvested and the intracellular lipid deposition was assessed using the corresponding kit and 2,7-dichlorofuorescin diacetate (DCFH-DA) fluorescent probe Level and oxidative stress. The gene and protein expressions of key molecules involved in lipid metabolism were detected by PCR and Western blot, respectively. Results: Compared with Pa group, oxymatrine and metformin significantly reduced triglyceride (TG) content (POmt = 0.023, PMet = 0.043) and upregulated the activity of carnitine acyl transferase -1 (POmt = 0.000, PMet = 0.000), and m RNA of fatty acid translocase (FAT / CD36) (POmt = 0.000, PMet = 0.000). The fluorescence intensity of DCFH-DA, total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH- Px) and malondialdehyde (MDA) showed that the drug intervention could significantly increase the activity and reduce the activity of T-SOD (POmt = 0.007, PMet = 0.007) and GSH-Px (POmt = 0.183, PMet = Generation of reactive oxygen species (ROS) and MDA (POmt = 0.016, PMet = 0.001). Conclusion: Oxymatrine can improve lipid-induced intracellular lipid deposition in HepG2 cells, which may be through the improvement of lipid metabolism and intracellular oxidative stress.