目的 研究肾必宁颗粒对IgA肾病(Immunoglobulin A nephropathy,IgAN)大鼠肾脏保护及对肾组织Toll样受体4/髓样分化因子88/核转录因子-KB(TLR4/MyD88/NF-KB)表达的影响.方法 将42只SD大鼠随机分为空白组、模型组、肾必宁低剂量组、肾必宁高剂量组、泼尼松组.除空白组外建立IgAN大鼠模型,从建模第4周开始,造模组接受相应药物或生理盐水灌胃,于第17周末麻醉后取材,全自动生化分析仪检测24小时尿蛋白定量(24hUTP)、尿红细胞计数(URBC)、血清白蛋白(ALB)、谷丙转氨酶(ALT)、血肌酐(Scr)、血尿素氮(BUN),光镜观察肾组织病理形态改变,免疫荧光观察系膜区IgA沉积情况;ELISA法检测血清中肿瘤坏死因子α(TNF-α)及白介素-6(IL-6)的含量;Western blot、荧光定量PCR法分别检测肾脏组织中TLR4、MyD88、NF-KB蛋白和mRNA表达.结果 与空白组比较,模型组大鼠系膜细胞增生、大量肾小管扩张,小管上皮细胞坏死,IgA电子致密物沉积增多,呈不规则颗粒及团块状分布;ALB水平降低,24hUTP、URBC、ALT、Scr和BUN水平均升高(P＜0.05或P＜0.01);血清TNF-α及IL-6含量显著升高(P＜0.01);肾组织中TLR4、MyD88和NF-κB蛋白及mRNA的表达显著升高(P＜0.01).与模型组比较,泼尼松组、肾必宁颗粒各组大鼠肾脏病理减轻,IgA电子致密物沉积减少;ALB水平升高,24hUTP、URBC、ALT、Scr和BUN水平均降低(P＜0.05或P＜0.01);血清TNF-α及IL-6含量显著降低(P＜0.01);肾组织中TLR4、MyD88和NF-KB蛋白及mRNA的表达均显著降低(P＜0.01).结论 肾必宁颗粒能有效改善IgAN肾脏损伤,可能通过调控TLR4/MyD88/NF-κB信号通路抑制炎症因子释放发挥其治疗作用.
Effects of Shenbining Granules on Renal Protection and TLR4/MyD88/NF-κB Signal Pathway in IgA Nephrotic Rats
Objective To study the effects of Shenbining granules on the renal protection of IgA nephropathy(IgAN)rats and toll like receptor 4/myeloid differentiation factor 88/nuclear transcription factor(TLR4/MyD88/NF-κB)signaling pathway.Meth-ods 42 SD rats were randomly divided into blank group,model group,low-dose group of Shenbining,high-dose group of Shenbining and prednisone group.Establishment of IgAN rat models except for the blank group.From the 4th week of modeling,the model rats received the corresponding drug intragastric intervention.At the end of the 17th week,the samples were taken after anesthesia.The 24 hour urinary protein(24hUTP),urinary red blood cell count(URBC),serum albumin(ALB),alanine transaminase(ALT),blood creatinine(Scr),and blood urea nitrogen(BUN)were detected by automatic biochemical analyzer,and the pathological changes of renal tissue were observed by light microscope,Immunofluorescence observation of IgA deposition in the mesangial area;The content of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in serum were detected u-sing ELISA;The expression levels of TLR4,MyD88,NF-κB proteins,and mRNA in kidney tissue were detected using Western blot and fluorescence quantitative PCR techniques,respectively.Results Compared with the blank group,the model group showed proliferation of mesangial cells,extensive dilation of renal tubules,necrosis of tubular epithelial cells,and increased deposition of IgA electron dense substances,distributed in irregular granules and clumps;The ALB level decreased,while the 24-hour UTP,URBC,ALT,Scr,and BUN levels all increased(P＜0.05 or P＜0.01);The levels of TNF-α and IL-6 in serum significant-ly increase(P＜0.01);The expression of TLR4,MyD88,NF-κB protein and mRNA in renal tissue significantly increased(P＜0.01).Compared with the model group,the pathological changes in the kidneys of rats in the prednisone group and Shenbining granules groups were alleviated,and the deposition of IgA electron dense substances was reduced;The level of ALB increased,while the levels of 24-hour UTP,URBC,ALT,Scr,and BUN decreased(P＜0.05 or P＜0.01);The levels of TNF-α and IL-6 in serum significantly reduced(P＜0.01);The expression of TLR4,MyD88,NF-κB protein and mRNA in renal tissue significantly reduced(P＜0.01).Conclusion Shenbining granules can effectively improve IgAN renal injury,which may play a role by regulating the TLR4/MyD88/NF-κB signaling pathway to inhibit the release of inflammatory factors.
IgA nephropathyShenbining GranulesTLR4/MyD88/NF-κB signaling pathway
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