芪黄健脾滋肾颗粒调控补体C1q影响巨噬细胞极化缓解系统性红斑狼疮小鼠肾损伤OA

Objective:To study the effect of Qihuang Jianpi Zishen(芪黄健脾滋肾)Granules on macrophage po-larization in the mouse model of systemic lupus erythematosus(MRL/lpr)based on complement 1q(C1q).Methods:MRL/lpr mice were randomized into model,Qihuang Jianpi Zishen Granules(3.9 g/kg),prednisone acetate(3 mg/kg),and C1q inhibitor(40 U/kg)groups(n=6),and C57BL/6 mice were included in the blank control group.The drugs were administered at a fixed time daily for 8 weeks.Pathological changes of the renal tissue were observed by HE staining,PAS staining,and Masson staining,and the collagen volume fraction(CVF)was calculated.The levels of creatinine(CREA),blood urea nitrogen(BUN),and urinary protein(PRO)were measured by biochemical assay kits.Enzyme-linked immunosorbent assay was employed to determine the levels of C1q,interleukin(IL)-10,transforming growth factor β1(TGFβ1),tumor necrosis factor-α(TNF-α),and IL-6 in the renal homogenate.The protein and mR-NA levels of arginase 1(Arg1),IL-10,IL-6,and inducible nitric oxide synthase(iNOS)in the renal tissue were deter-mined by Western blot and RT-qPCR,respectively.The proportions of CD86+M1 and CD206+M2-type macrophages in the renal tissue were determined by flow cytometry.The levels of M1-type macrophages expressing F4/80+iNOS and M2-type macrophages expressing F4/80+Arg1 were determined by the double immunofluorescence labeling method.Results:Compared with the blank control group,the model group showed pathological changes such as glomerular basement mem-brane thickening,partial renal interstitial edema,and inflammatory cell infiltration,elevated levels of CVF,PRO,CREA,BUN,anti-ds-DNA antibody,TNF-α,and IL-6(P<0.01),declined levels of C1q,TGFβ1,and IL-10(P<0.01),up-reg-ulated protein and mRNA levels of IL-6 and iNOS(P<0.01),down-regulated protein and mRNA levels of Arg1 and IL-10(P<0.01),and increased M1/M2 ratio(P<0.01)due to the increased proportion of M1-type macrophages and the decreased proportion of M2-type macrophages(P<0.01).Compared with the model group,Qihuang Jianpi Zishen Gran-ules(3.9 g/kg)ameliorated the pathological changes and recovered the glomerular structure and tubular arrangement in the renal tissue,lowered the levels of CVF,PRO,CREA,BUN,anti-ds-DNA antibody,TNF-α,and IL-6(P<0.01),raised the levels of C1q,TGFβ1,and IL-10(P<0.05),down-regulated the protein and mRNA levels of IL-6 and iNOS(P<0.01),up-regulated the protein and mRNA levels of Arg1 and IL-10(P<0.01),and decreased the M1/M2 ratio(P<0.01)by reducing the proportion of M1 macrophages and increasing the proportion of M2 macrophages(P<0.01).Conclusion:Qihuang Jianpi Zishen Granules can protect the kidney of MRL/lpr mice by regulating macrophage polariza-tion via C1q.