当归芍药散通过调节db/db小鼠肾脏巨噬细胞极化减轻糖尿病肾病炎症反应的作用机制OA

Objective:To observe the effects of Danggui Shaoyaosan on macrophage polarization and renal inflammation in db/db mice with diabetic kidney disease(DKD),and to explore its renal protective effects and underlying mechanisms.Methods:Eight db/m mice were assigned to the normal group,and forty db/db mice were randomly divided into a model group,low-,medium-,and high-dose Danggui Shaoyaosan groups(8.39,16.77,33.54 g·kg-1),and an irbesartan group(0.025 g·kg-1).All mice were administered treatment by gavage for 12 consecutive weeks.General conditions of the mice were observed during the intervention.At the end of the 12-week intervention,24-h urine samples were collected using metabolic cages,after which the mice were anesthetized for sample collection.Blood was collected by enucleation and centrifuged to obtain serum for the determination of glycated serum protein(GSP),serum creatinine(SCr),blood urea nitrogen(BUN),total cholesterol(TC),and triglycerides(TG).The urinary albumin-to-creatinine ratio(UACR)was measured.Renal pathological changes were observed using hematoxylin-eosin(HE)staining,periodic acid-Schiff(PAS)staining,and Masson staining.Enzyme-linked immunosorbent assay(ELISA)was used to detect serum tumor necrosis factor-α(TNF-α),interleukin-10(IL-10),and monocyte chemoattractant protein-1(MCP-1)levels.Immunofluorescence(IF)was performed to detect F4/80 expression in renal tissue,and immunohistochemistry(IHC)was used to assess CD206 expression.Real-time quantitative polymerase chain reaction(Real-time PCR)was employed to measure the mRNA expression of TNF-α,IL-10,inducible nitric oxide synthase(iNOS),and arginase-1(Arg-1).Western blot analysis was used to detect the protein expression of iNOS,Arg-1,CD86,and CD206 in renal tissue.Results:Compared with the normal group,the model group showed increased levels of GSP,UACR,SCr,BUN,TC,and TG,elevated levels of the inflammatory factor TNF-α and the chemokine MCP-1,and decreased IL-10 levels(P＜0.01).Pathological examination revealed glomerular hypertrophy,mesangial cell proliferation with marked mesangial expansion,inflammatory cell infiltration,vacuolar degeneration of renal tubular epithelial cells,prominent glycogen deposition,and increased collagen fiber deposition.In addition,relative F4/80 fluorescence intensity was enhanced,CD206 expression in the glomeruli and renal interstitium was reduced,and TNF-α and iNOS mRNA expression was increased.IL-10 and Arg-1 mRNA expression was decreased,iNOS and CD86 protein expression was increased,and Arg-1 and CD206 protein expression was decreased(P＜0.05,P＜0.01).Compared with the model group,the Danggui Shaoyaosan groups and the irbesartan group showed decreased levels of GSP,UACR,SCr,BUN,TC,and TG,reduced serum TNF-α and MCP-1 levels,and increased IL-10 levels.Renal pathological damage was improved to varying degrees.Relative F4/80 fluorescence intensity was reduced,CD206 expression in the glomeruli and renal interstitium was increased,and TNF-α and iNOS mRNA expression was decreased.IL-10 and Arg-1 mRNA expression was increased,iNOS and CD86 protein expression was reduced,and Arg-1 and CD206 protein expression was increased(P＜0.05,P＜0.01).Conclusion:Danggui Shaoyaosan can improve renal function and alleviate renal pathological damage in db/db mice.Its mechanism may be related to inhibiting Ml pro-inflammatory macrophage polarization,promoting M2 anti-inflammatory macrophage polarization,reducing inflammatory responses,delaying the progression of renal fibrosis,improving renal pathological injury,and thereby exerting renal protective effects.