从肺肾同治探讨扶正祛风方经miRNA-30/TGF-β1/Smad保护足细胞防治特发性膜性肾病的作用及机制

Idiopathic membranous nephropathy (IMN) is characterized by podocyte injury. PM2.5 increases the risk of IMN. miRNA-30/TGF-β1/Smad iduces podocyte apoptosis as well as PM2.5-induced pulmonary impairment. We create Fuzhengqufeng Formula (composition: Astragalus, Dioscorea nipponica, Herba Siegesbeckiae, etc.). Our previous studies have confirmed the efficiency,including alleviate respiratory symptoms and down-regulating TGF-β1 in IMN rat models. We consider podocyte associated with lung in IMN kidney and suppose that Fuzhengqufeng Formula inhibit podocyte apoptosis and allivate PM2.5-induced pulmonary impairment via miRNA-30/TGF-β1/Smad. In this study, we would firstly treat C-BSA rat and PM2.5-induced pulmonary impairment model by Fuzhengqufeng Formula and confirm the regulation of miRNA-30/TGF-β1/Smad, podocyte apoptosis-related proteins and EMT symbol of lung. Secondly, we would establish podocyte injury model induced by C5b-9, with the intervention by rat serum of PM2.5. Then we would treat the model by Fuzhengqufeng Formula, and explore the influence of miRNA-30/TGF-β1/Smad and podocyte apoptosis-related proteins in vitro. The findings of this study will help to understand the mechanism of podocyte protection in IMN by Fuzhengqufeng Formula treatment as a form of treating liver and kidney simultaneously in TCM.

特发性膜性肾病（IMN）以足细胞损伤为病理特点，细颗粒物可增加发病风险。miRNA-30/TGF-β1/Smad是介导IMN足细胞凋亡及细颗粒物肺损伤的共同机制。我们创制扶正祛风方（组成：生黄芪、穿山龙、豨莶草等），发现该方改善IMN患者肺系症状，前期研究证实可下调足细胞TGF-β1表达。我们提出IMN肾小球足细胞与肺相关，假设该方通过miRNA-30/TGF-β1/Smad抑制足细胞凋亡、减轻细颗粒物肺损伤。本研究应用C-BSA大鼠模型及霾颗粒肺损伤大鼠模型，观察该方对足细胞miRNA-30/TGF-β1/Smad及凋亡相关蛋白、肺EMT表型标志物表达的影响；构建C5b-9诱导的足细胞亚溶破模型，以吸入雾霾颗粒大鼠血清干预足细胞，研究该方含药血清对足细胞miRNA-30/TGF-β1/Smad及凋亡相关蛋白表达的影响，从而探讨扶正祛风方基于肺肾同治保护足细胞防治IMN的科学内涵。

特发性膜性肾病（IMN）以足细胞损伤为病理特点，PM2.5可增加发病风险。miR-30/TGF-β1/Smad是介导足细胞凋亡及PM2.5肺损伤的共同机制。本研究应用C-BSA大鼠模型及PM2.5肺损伤大鼠模型，观察扶正祛风方对肺和肾miR-30/TGF-β1/Smad及凋亡标志物、肺EMT标志物的影响；构建吸入PM2.5大鼠血清干预及C5b-9诱导的足细胞损伤模型，研究该方含药血清对足细胞miR-30/TGF-β1/Smad及凋亡标志物的影响。研究发现PM2.5慢染毒导致大鼠肺组织损伤，增加EMT标志物α-SMA表达；加重C-BSA大鼠蛋白尿和肾组织损伤，提高足细胞凋亡率，减少足细胞特异性蛋白WT1表达；增加肺和肾组织TGF-β1、Smad2、Smad3、促凋亡蛋白Bax和Caspase-3表达，减少miR-30、Smad7和抑凋亡蛋白Bcl-2表达。扶正祛风方有效降低PM2.5慢染毒C-BSA大鼠蛋白尿，减轻肾组织损伤，降低足细胞凋亡率，上调WT1表达；有效减轻肺组织损伤，下调α-SMA表达；下调肺和肾组织TGF-β1、Smad2、Smad3 、Bax和Caspase-3表达，上调miR-30、Smad7和Bcl-2表达。吸入PM2.5大鼠血清干预及C5b-9诱导的足细胞损伤模型细胞凋亡率升高，TGF-β1、Smad2、Smad3、Bax、Caspase-3表达增加，miR-30、Smad7、Bcl-2和WT1表达减少。扶正祛风方含药血清有效降低足细胞损伤模型凋亡率，下调TGF-β1、Smad2、Smad3、Bax、Caspase-3表达，上调miR-30、Smad7、Bcl-2和WT1表达。提示扶正祛风方通过miR-30/TGF-β1/Smad抑制PM2.5环境下IMN足细胞凋亡，减轻肺损伤，阐释了扶正祛风方基于肺肾同治保护足细胞防治IMN的科学内涵。