Tanshinone Ⅰ Relieves Hepatic Ischemia-reperfusion Injury By Activating Nrf2/HO-1 Pathway

Objective: Explore Tanshinone Ⅰ(Tanshinone I,T-I)in hepatic ischemia-reperfusion injury(hepatic ischemia reperfusion injury,HIRI)mouse model of protection and reveal its action mechanism.Methods: Get 36 C57 mice ready,All 8 weeks old,Using random methods,Divide it into six groups,(sham,)Impositional surgery n=6,ischemia-reperfusion(IR,)group n=6,IR T-I(40 mg/kg,)n=6,IR T-I(20mg/kg,)n=6,IR T-I group(10 mg/kg,)n=6 and IR T-I groups(5 mg/kg,)n=6).All groups were intraperitoneally injected with olive oil.IR group + T-I dissolved in olive oil injection of different concentration of tanshinone Ⅰ,dosing 1 times daily,for seven days in a row,the last for 2 hours after 70% of liver ischemia-reperfusion injury model was established,after anesthesia,into the abdomen,in mechanical way,the first liver door left,the middle of the portal vein and hepatic artery branch clamp,form 70% of ischemia,reperfusion in mice after 6 hours of euthanasia,and collect the peripheral blood and liver tissue samples;According to the kit instructions,Detection of serum ALT、AST,to reflecting the level of liver function;The indexes of reduced malondialdehyde(MDA),superoxide dismutase(SOD),glutathione(GSH),and Caspase-3 in liver also need to be tested,to reflect oxidative stress levels;By using TUNEL apoptosis detection method,To reveal the apoptosis of hepatocytes;HE staining to evaluate the pathological injury of liver tissue;Immunohistochemical method was used to detect the expression of heme oxygenase-1(HO-1)and Caspase-3 protein,Western Blot detection of Nrf2 and HO-1 gene expression levels,Analyze the effect of Nrf2/HO-1 pathway on liver ischemia reperfusion injury.Metrology data are characterized as mean ±standard deviation.Enter data and data into SPSS22.0 software for processing and analysis,The metrological data were compared by one factor ANOVA,Comparison between the two groups was done through LSD-t tests,P <0.05 represented a significant difference.Results:(1)T-I alleviated HIRI in mice.Serum ALT and AST in the IR group were significantly higher than those in the sham group(P <0.01),there were significant differences between the two groups.IR+T-I(20mg/kg)group was significantly lower than IR group(P<0.01),and the difference was statistically significant.Therefore,20mg/kg T-I was determined as the best treatment concentration.He staining showed focal or large area degeneration and necrosis of liver cells and hepatic lobular structure disorder in the IR group.Compared with the IR group,the area of cell necrosis in the IR T-I group(20mg/kg)decreased significantly and the liver tissue structure was basically intact;(2)T-I enhance liver antioxidant capacity in HIRI mouse models.On the SOD and GSH indicators,IR T-I group(20mg/kg)was significantly higher than IR group(P<0.05),But at the MDA level,IR T-I group(20mg/kg)was significantly lower(P<0.05),There was significant difference between the two groups;(3)T-I inhibition of hepatocyte apoptosis in HIRI mouse model.As an indicator of Caspase-3 activity,IR group was significantly higher than sham group(P<0.01),There was significant difference between the two groups;IR T-I group(20mg/kg)was significantly lower than IR group(P<0.01),There was significant difference between groups.Detection of Caspase 3,by immunohistochemistry Found that by IR group,IR T-I(20mg/kg)group Caspase-3 protein expression level is lower;According to the TUNEL results,In terms of hepatocyte apoptosis,Compared to IR group,IR T-I(20mg/kg)group was more optimistic.(4)T-I attenuates HIRI.by activating Nrf2/HO-1 pathways The Western Blot results showed that IR group was significantly higher than sham group,but lower than 20mg/kg group.Conclusion: T-I attenuates hepatic ischemia-reperfusion injury by activating Nrf2/HO-1 signaling pathway,reducing hepatocyte apoptosis and inhibiting oxidative stress.