基于Th17/Treg细胞免疫平衡探讨升阳益胃汤治疗特发性膜性肾病的作用及机制

Study on the effect and mechanism of Shengyang Yiwei Decoction in treating idiopathic membranous nephropathy based on the immune balance of Th17/Treg cells

关键词:  升阳益胃汤  膜性肾病  足细胞  免疫平衡  Th17/Treg

Key words:  Shengyang Yiwei Decoction  membranous nephropathy  podocyte  immune balance  Th17/Treg

摘要:

[目的] 探讨升阳益胃汤对膜性肾病（MN）大鼠模型的保护作用及机制。[方法] 将40只雄性SD大鼠随机分为对照组10只和造模组30只，造模组尾静脉注射羊抗大鼠FX1A血清制备海曼（Heymann）肾炎模型。将造模成功的大鼠随机分为模型组、升阳益胃汤组和盐酸贝那普利组各10只，并予以相应药物灌胃。干预6周后，检测24 h尿蛋白（24 h-UTP）、总蛋白（TP）、白蛋白（Alb）、血清总胆固醇（TC）、三酰甘油（TG）、肌酐（Scr）、尿素氮（BUN）；取肾组织行苏木素-伊红（HE）、马松（Masson）和六胺银（PASM）染色，观察肾组织病理形态变化，蛋白免疫印迹法（Western blot）法检测肾组织白细胞介素-17A（IL-17A）、白细胞介素-10（IL-10）、α-辅肌动蛋白-4（α-actinin-4）、足糖萼蛋白[Podocalyxin（Podxl）]表达情况；取大鼠脾脏检测Th17和Treg细胞数量。[结果] 与对照组相比，模型组大鼠24 h-UTP、TC、TG水平显著升高，TP、Alb水平显著降低（P<0.01），Scr、BUN水平差别无统计学意义（P>0.05）；模型组肾脏病理改变明显；α-actinin-4、Podxl表达均明显减少（P<0.01、P<0.05），IL-17A水平显著升高，IL-10水平明显降低（P<0.01）；Th17％、Th17/Treg显著升高，Treg％显著降低（P<0.01）。与模型组相比，各治疗组24 h-UTP、TC、TG、TP水平显著降低（P<0.05或P<0.01），Alb水平明显升高（P<0.05）；肾脏病理改变减轻。其中，Z组α-actinin-4、Podxl表达明显增加（P<0.05或P<0.01）；IL-17A、Th17％、Th17/Treg水平明显降低，IL-10、Treg％水平升高（P<0.05或P<0.01）。[结论] 升阳益胃汤可降低膜性肾病大鼠蛋白尿，减轻肾脏病理损伤，并提高足细胞α-actinin-4、Podxl的表达，具有明显的肾脏保护作用，其作用机制可能与调节Th17/Treg免疫平衡有关。

Abstract:

[Objective] Exploring the protective effect and mechanism of Shengyang Yiwei Decoction on membranous nephropathy(MN) rat model. [Methods] Forty male SD rats were randomly divided into control group with 10 rats and the modeling group with 30 rats. Rats were iv sheep anti-rat FX1A serum by tail to establish Heymann nephritis model. The rats with successful modeling were randomly divided into model group,Shengyang Yiwei Decoction group and Benazepril hydrochloride group,with 10 rats in each group,and the corresponding drugs were given by gavage. After 6 weeks of intervention,24 h-UTP,TP,Alb,TC,TG,Scr and BUN were detected,while renal tissue was stained with HE,Masson and PASM to observe the pathological changes of renal tissue,and Western blot method was used to detect the expression of IL-17A,IL-10,α-actinin-4,and Podocalyxin(Podxl) in renal tissue. Meanwhile,rat spleen was taken to detect the quantity of Th17 and Treg cells. [Results] Compared with the C group,the 24 h-UTP,TC,TG levels of rats in the M group were significantly higher,and TP and Alb levels were significantly lower(P<0.01),whereas the differences in Scr and BUN levels were not statistically significant(P>0.05). Meanwhile,renal pathological changes were obvious in the M group,and α-actinin-4 and Podxl expression were significantly decreased(P<0.01,P<0.05). Additionally,IL-17A significantly increased and IL-10 decreased(P<0.01). And Th17% and Th17/Treg were significantly higher and Treg% was significantly lower(P<0.01). Compared with the M group,the levels of 24h-UTP,TC,TG,and TP were significantly reduced(P<0.01,P<0.05) and the level of Alb was significantly increased(P<0.05) in each treatment group. Simultaneously the pathological changes of kidney were improved. Among them,the expression of α-actinin-4 and Podxl was significantly increased in the Z group(P<0.05,P<0.01),while IL-17A,Th17% and Th17/Treg were significantly reduced,and IL-10 and Treg% were increased(P<0.01,P<0.05). [Conclusion] Shengyang Yiwei Decoction can reduce proteinuria,attenuate renal pathological injury and increase the expression of α-actinin-4 and Podxl in podocytes expressing,which has obvious renal protective effects in rats with membranous nephropathy. In addition,the mechanism of these protective effects may be related to the regulation of Th17/Treg immune balance.