基于IL-6/JAK2/STAT3信号通路探讨海昆肾喜胶囊治疗慢性肾脏病肾纤维化作用机制OA

Objective To investigate the mechanism of Haikun Shenxi Capsules in attenuating renal fibrosis in chronic kidney disease(CKD)based on IL-6/JAK2/STAT3 signaling pathway.Methods Totally 48 male C57BL/6 mice were randomly divided into the blank group,model group,Haikun Shenxi Capsules group(200 mg/kg)and losartan potassium group(10 mg/kg).Except for the blank group,mice in the other groups were fed a 0.2%adenine-containing diet for 4 weeks to induce CKD.Mice in the treatment groups received the corresponding drugs by daily gavage from the start of model induction,once a day for 8 weeks.The levels of serum creatinine,blood urea nitrogen,serum uric acid,alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were detected;renal index and urinary albumin creatinine ratio(ACR)were detected;HE,Masson and Sirius red staining were used to observe the renal morphology and fibrosis,renal ultrastructure was observed by transmission electron microscopy,the expressions of Fibronectin and E-cadherin were detected by immunofluorescence,the key targets and pathways were screened by transcriptomics,and the contents of interleukin(IL)-6,IL-1β and tumor necrosis factor(TNF)-α in renal tissue were detected by ELISA,the mRNA expressions of IL-6,JAK2,STAT3 were detected by RT-qPCR,and the protein expressions of IL-6,JAK2,p-JAK2,STAT3 and p-STAT3 were detected by Western blot.Results Compared with the blank group,the levels of serum creatinine,blood urea nitrogen,serum uric acid,ALT,renal index and ACR in the model group significantly increased(P<0.05,P<0.01);the renal tissue structure was damaged,which showed that the renal tubular lumen was dilated,the brush border fell off,the interstitial collagen fibers proliferated,and the collagen deposition increased significantly(P<0.01);the renal tubular epithelial cells were necrotic,cytoplasmic electron density decreased,vacuolation and nucleolysis,the expression of Fibronectin was significantly increased,and the expression of E-cadherin significantly decreased(P<0.01).Compared with the model group,the levels of serum creatinine,blood urea nitrogen,blood uric acid,renal index and ACR in Haikun Shenxi Capsules group and losartan potassium group significantly decreased(P<0.05,P<0.01);the vacuolation,lumen expansion and collagen deposition of renal tubules were improved,the expression of Fibronectin significantly decreased,and the expression of E-cadherin significantly increased(P<0.05,P<0.01).Transcriptomic sequencing showed that Haikun Shenxi Capsules could regulate 295 key differentially expressed genes related to CKD that were primarily enriched in the JAK-STAT signaling pathway.ELISA showed that both Haikun Shenxi Capsules and losartan potassium could significantly reduce the contents of renal IL-6,IL-1β and TNF-α(P<0.05,P<0.01).RT-qPCR showed that both treatments reduced the mRNA expression of IL-6 and STAT3(P<0.05,P<0.01).Western blot analysis further showed that both treatments could suppress IL-6 protein expression and the phosphorylation of JAK2 and STAT3(P<0.05,P<0.01).Conclusion Haikun Shenxi Capsules may delay CKD progression by inhibiting the IL-6/JAK2/STAT3 signaling pathway,reducing inflammatory cytokine levels,and thereby attenuating renal inflammation and fibrosis.