重组旋毛虫半胱氨酸蛋白酶抑制剂在脓毒症急性肝损伤治疗中的研究OA

Objective:To investigate the potential role of recombinant trichinella spiralis cystatin protease inhibitor(r TS-Cys)in the treatment of acute liver injury in sepsis.Methods:Male C57BL/6 mice were randomly divided into 7 groups(n=10)[Sham group,CLP group,CLP+r TS-Cys group(including 10,20,30,40 μg),CLP+DEX group].The CLP and CLP+r TS-Cys groups were used to establish a mouse model of sepsis-induced acute liver injury by cecal ligation and puncture.The Sham group underwent laparotomy without other treatment,and the abdomen was closed immediately.One hour later,100 μL PBS was intraperitoneally injected in the CLP group mice,and 100 μL PBS containing different doses r TS-Cys was intraperitone-ally injected in the CLP+r TS-Cys groups mice,and the same amount of dexamethasone was used in the CLP+DEX group.After 24 hours,the mice were sacrificed and the pathological changes of the liver were evaluated by H&E staining.The expression lev-els of TNF-α,IL-6,IL-10 and TGF-β in serum were detected by ELISA.The levels of ALT and AST in serum were detected by automatic biochemical analyzer.The expression of NF-κB/MAPK signaling pathway proteins was detected by Western blot.Results:Survival analysis suggested that r TS-Cys could improve the survival rate of septic mice,The 20 μg group of mice exhibit-ed the highest survival rate at 72 h post-operation and proceeded to the subsequent experiments.H&E staining results showed that the liver tissue structure of the CLP group was disordered and inflammatory cell infiltration was observed.Compared to the CLP group,the pathological damage of the liver tissue in the CLP+r TS-Cys group and CLP+DEX group was alleviated,and the in-flammatory cell infiltration was significantly reduced.ELISA analysis demonstrated distinct patterns in inflammatory markers.Se-rum TNF-α and IL-6 levels were significantly elevated in the CLP group compared to the Sham group,with subsequent reduction observed in the CLP+r TS-Cys group and CLP+DEX group.Conversely,anti-inflammatory cytokines IL-10 and TGF-β showed progressive increases across groups,with CLP levels exceeding Sham values,and CLP+r TS-Cys group and CLP+DEX group exhibiting the highest levels.Liver function parameters,including ALT and AST,were markedly elevated in the CLP group com-pared to Sham controls,while treatment with r TS-Cys and DEX effectively lowered these enzyme levels below those observed in the CLP group.Western blot results showed that the expression of NF-κB/MAPK signaling pathway protein was up-regulated in the CLP group,and the expression of this pathway in the CLP+r TS-Cys group was lower than that in the CLP group.Conclu-sion:rTS-Cys can inhibit the expression of pro-inflammatory cytokines and promote the expression of regulatory cytokines by down-regulating the NF-κB/MAPK signaling pathway,thereby protecting sepsis-induced acute liver injury.