复方美洲大蠊胶囊通过调节JAK2/STAT3信号通路平衡Th17/Treg轴缓解大鼠湿热泄泻OA

Objective:This paper aims to evaluate the therapeutic efficacy of the compound formula capsules of Periplaneta americana on dampness-heat diarrhea(DHD)in rats and investigate its effects on the JAK2/STAT3 signaling pathway.Methods:DHD model was established through a high-sugar and high-fat diet,heterologous antigen sensitiza-tion,and 2,4,6-trinitrobenzene sulfonic acid(TNBS)enema.Rats were divided into model control,positive control group treated with Xianglian Pills at 300 mg/kg,groups treated with compound formula capsules of Periplaneta americana at 283 mg/kg and 566 mg/kg,and a normal control group.After 14 days of treatment,the following indexes were as-sessed:rat general condition,body weight,fecal pellet count,urine volume,and rectal temperature,as well as food and water intake were monitored by using metabolic cages combined with traditional Chinese medicine syndrome scoring.The disease activity index(DAI),organ index,and colonic mucosal damage index(CMDI)were calculated.The colonic pa-thology was observed and scored via hematoxylin-eosin(HE)staining.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of IL-6,IL-8,COX-2,TNF-α,and CRP in the serum,as well as the contents of TGF-β,MPO,IL-2,IL-17,VEGF,bFGF,and PDGF in colon tissues.The protein expressions of IL-6,JAK2,P-JAK2,STAT3,P-STAT3,and SOCS3 in colon tissues were detected by western blot.Flow cytometry was used to detect the expression of T cell subsets(CD3+,CD4+,and CD8+)and intracellular cytokines(IFN-γ,IL-4,IL-17,and Foxp3)in the spleen and mesenteric lymph nodes.Results:Compared with those in the normal control group,rats in the model control group ex-hibited typical damp-heat diarrhea symptoms.The indices of spleen,liver,kidney,and colon were significantly increased(P<0.01),while CMDI and HS pathological scores were significantly elevated(P<0.01).The contents of IL-6,IL-8,COX-2,TNF-α,and CRP in serum,as well as the contents of MPO,IL-2,IL-17,VEGF,bFGF,and PDGF in colon tissues were significantly elevated(P<0.01),while TGF-β contents in colon tissues were significantly reduced(P<0.01).The IFN-γ and IL-17 contents in spleen and mesenteric lymph nodes were significantly increased(P<0.05 or P<0.01),while IL-4 and Foxp3 contents were significantly reduced(P<0.05).The phosphorylation expression of the JAK2/STAT3 pathway was significantly up-regulated(P<0.01),while SOCS3 protein expression was significantly down-regula-ted(P<0.05).Compared with the model control group,groups with different doses of Compound formula capsules of Periplaneta americana showed significantly improved symptoms in DHD rats.In the group treated at 566 mg/kg,the DAI scores,the indices of liver,kidney,and colon organ,CMDI,as well as HS pathological scores were significantly reduced(P<0.01).The contents of IL-6,IL-8,COX-2,TNF-α,and CRP in serum,as well as MPO,IL-2,IL-17,VEGF,bFGF,and PDGF in colon tissues were significantly reduced(P<0.01),while the content of TGF-β in colon tissues was signifi-cantly increased(P<0.01).The IFN-γ and IL-17 contents in the spleen and mesenteric lymph nodes were significantly decreased(P<0.05 or P<0.01),while IL-4 and Foxp3 contents were increased(P<0.05).GF and PDGF contents were significantly reduced(P<0.01),while TGF-β content in colon tissues was increased(P<0.01).The proportion of IL-17+CD4+T cells and IL-17/IFN-γ contents were decreased in the spleen and mesenteric lymph nodes(P<0.05 or P<0.01),while the proportion of Foxp3+CD4+T cells was increased(P<0.05).The phosphorylation expression of the JAK2/STAT3 pathway was significantly down-regulated(P<0.01),while SOCS3 protein expression was up-regulated(P<0.05).Conclusion:The compound formula capsules of Periplaneta americana alleviates DHD by inhibiting JAK2/STAT3 activation,rebalancing Th17/Treg immunity,and suppressing inflammatory responses.