Mechanism of oxidative stress induced by Gynura japonica to abnormal liver function in rats
Objective To investigate the oxidative stress mechanism of liver function abnormality in rats caused by Gynu-ra japonica exposure,so as to provide a theoretical basis for clinical treatment and prevention.Methods(i)For the ani-mal experiments,24 female SD rats were randomly divided into 2 groups:a treatment group and a control group,each group had 12 rats.In the treatment group,Gynura japonica decoction was administered by intragastric administration at a dose of 15 000 mg/kg,with a gavage volume of 10 mL/(kg·bw)for 1 time/d.The control group was gavaged with puri-fied water and fed normally.The 2 groups were executed on the 7th and 14th days,respectively.Blood was collected from the abdominal aorta and the blood biochemical indexes,including AST,ALT,ALP and ALB,were detected.The coagula-tion indexes,including APTT and PT,were also detected.The livers were extracted and observed using HE staining to de-tect pathological manifestations of the liver tissue.Finally,a tissue homogenizer was used to observe the liver tissue.The liver tissue was processed with a tissue homogeniser,and the supernatant was extracted and tested for malondialdehyde(MDA),superoxide dismutase(SOD),catalase(CAT)and glu-tathione peroxidase(GPX)using a sunrise enzyme marker.(ii)For the in vitro experiments,the rat hepatocytes were cultured with the following groups:blank control group,40 mg/mL Gynura japonica group,20 mg/mL Gynura japonica group,10 mg/mL Gynura japonica group,and 5 mg/mL Gynura japon-ica group.Cells were incubated in DMEM,40 mg/mL Gynura japonica,20 mg/mL Gynura japonica,10 mg/mL Gynura ja-ponica and 5 mg/mL Gynura japonica for 2 hours.The level of intracellular reactive oxygen species in hepatocytes was de-tected by flow cytometry,as well as the level of intracellular reactive oxygen species in hepatocyte mitochondria,using the Mitochondria SOX probe and flow cytometry.Results(i)The results of the animal experiments:at 14 d of gavage,compared with the control group,serum ALT(t=19.707,P=0.000),AST(t=25.498,P=0.000)and ALP(t=21.452,P=0.000)were elevated in the treated group,while serum ALB(t=5.971,P=0.000)was decreased.Additionally,plasma PT(t=4.778,P=0.004)and APTT(t=4.884,P=0.004)were prolonged.The pathological results clearly showed significant inflammatory infiltration with necrosis in the liver of the treated group,and the injury worsened with time.At 14 d of gavage,the liver tissue homogenate of the treatment group showed increased MDA content(t=12.155,P=0.000)and decreased SOD expression(t=17.281,P=0.000),decreased CAT expres-sion(t=9.412,P=0.000),and decreased GPX activity(t=10.294,P=0.000).(ii)The in vitro experiments demon-strated that the intrahepatocyte and mitochondrial levels of reactive oxygen species were significantly elevated in the Gynu-ra japonica 10 mg/mL and 20 mg/mL and 40 mg/mL groups(F-values of 280.841 and 45.780,respectively,with P=0.000 in both cases).Conclusion Gynura japonica exposure causes liver function impairment in rats,and the mechanism may be related to the altered level of oxidative stress in vivo.
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