| Objective: To explore the effect relationship of hepatocyte ferroptosis in the process of ACLF and the therapeutic mechanism of Jiedu Huayu granule regulating SIRT1/Nrf2 signal to interfere with hepatocyte ferroptosis in ACLF.Methods: 75 SPF grade SD rats were randomly divided into five groups,15 rats in the control group(Group A),15 rats in the ACLF group(Group B),15 rats in JDHY group(Group C),15 rats in the ACLF+Res group(Group D),and 15 rats in the JDHY+Res group(Group E).The ACLF rat model was replicated with 50% carbon tetrachloride vegetable oil solution(CCl4: olive oil=1:1 ratio)+LPS/D-Gal N.The rats in the JDHY group and the JDHY+Res group were given 16.28/kg Jiedu Huayu granules by gavage 3 days before the model formation,twice a day,at an interval of 12 h,and lasted until 48 h after the model formation.The amount of gavage fluid was 2ml/100 g body weight per day.The other groups were given the same amount of distilled water instead.After modeling,serum and liver tissue of rats in each group were collected.HE and Masson staining were used to detect liver tissue pathology,biochemical indicators of AST,ALT and TBIL in serum of rats were detected by biochemical instrument,ferroptosis concentration was detected by enzyme-labeled instrument,and related pathway proteins SIRT1,Nrf2 and ferroptosis key proteins FTH1 and TfR1 were detected by Western blot.Results: The serum biochemical indexes showed that ALT,AST and TBIL in the ACLF group,the JDHY group,the ACLF+Res group and the JDHY+Res group were significantly higher than those in the control group(P < 0.05).Compared with the ACLF group,ALT,AST and TBIL in the JDHY group and the JDHY+Res group decreased significantly(P < 0.05),while there was no significant difference between the ACLF+Res group and the ACLF group(P >0.05).Compared with the JDHY group,ALT,AST and TBIL in the ACLF+Res group increased,while ALT,AST and TBIL in the JDHY+Res group decreased,with no significant difference.HE and Masson staining results showed that in the ACLF group,pseudolobules were formed in liver tissue,cells were arranged in disorder,liver cells were edema,degeneration and necrosis were obvious,severe inflammatory cell infiltration was accompanied by fibrous tissue proliferation,liver tissue was flaky necrosis and apoptosis;In the ACLF+Res group,the arrangement of hepatocytes around the central vein of liver tissue was disordered,pseudolobules were formed,the edema and degeneration of hepatocytes were alleviated,mild inflammatory cell infiltration was observed,and the number of bile ducts was decreased compared with that in the ACLF group;In the JDHY group,the hepatocytes around the central vein were more regular,with pseudolobule formation,edema and degeneration of hepatocytes were significantly reduced,and the number of inflammatory cells and bile ducts were decreased compared with that in the ACLF+Res group;The overall injury level of the JDHY+Res group was lower than that of the JDHY group.The results of iron content detection showed that compared with the control group,the iron content in the ACLF group,the JDHY group,the ACLF+Res group and the JDHY+Res group was significantly higher(P < 0.05);The iron content of the JDHY group,the ACLF+Res group and the JDHY+Res group was significantly lower than that of the ACLF group(P < 0.05);The iron content of the JDHY+Res group was lower than that of the JDHY group(P < 0.05).Western blot showed that the expression of SIRT1 and Nrf2 protein in liver tissue of the JDHY group,the ACLF+Res group and the JDHY+Res group was higher than that of the ACLF group(P < 0.05),while the expression of FTH1 and TFR1 protein was lower than that of the ACLF group(P < 0.05);The protein expression of SIRT1 in the ACLF+Res group and the JDHY+Res group was higher than that in the JDHY group(P < 0.05),and the expression of Nrf2 in the ACLF+Res group and the JDHY+Res group was higher(P <0.05).The difference in the expression of SIRT1 in the JDHY+Res group was statistically significant(P < 0.05).The protein expression of FTH1 and TFR1 in the ACLF+Res group and the JDHY+Res group was lower than that in the JDHY group(P < 0.05);Compared with the ACLF+Res group,the expression of SIRT1 and Nrf2 protein in liver tissue of the JDHY+Res group increased,and the expression of FTH1 and TFR1 decreased,with statistical significance(P < 0.05).Conclusion(s): Ferroptosis may be an important way of hepatocyte death in ACLF,and the occurrence of ferroptosis is not conducive to the prognosis and development of the disease.However,after the intervention of Jiedu Huayu granules,the expression of SIRT1/Nrf2 signal is up-regulated,the content of iron ions in hepatocytes is reduced,and the expression of ferroptosis related proteins is inhibited,suggesting that Jiedu Huayu granule antagonizing ACLF may inhibit the occurrence of hepatocyte ferroptosis by up-regulating the SIRT1/Nrf2 signal,Alleviate the pathological damage of liver tissue and improve the liver function of ACLF rats. |
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