益气活血通络方调控NR4A1/TGF-β1负反馈环路抑制单侧输尿管梗阻大鼠肾小管上皮间充质转化的机制  

Mechanism of Yiqi Huoxue Tongluo Recipe regulating NR4A1/TGF-β1 negative feedback loop to inhibit epithelial-mesenchymal transition in the renal tubules of UUO rats

作　　者：王洪双 何珍 张孟娟 方芳 刘佳志 王筝 钟艳 王香婷 WANG Hongshuang;HE Zhen;ZHANG Mengjuan;FANG Fang;LIU Jiazhi;WANG Zhenog;ZHONG Yan;WANG Xiangting(Hebei University of Chinese Medicine,Shijiazhuang 050200,China;Hebei Key Laboratory of Integrative Medicine on Liver-Kidney Patterns,Shijiazhuang 050091,China;Institute of Integrative Medicine,Hebei University of Chinese Medicine,Shijiazhuang 050091,China)

机构地区：[1]河北中医药大学,石家庄050200 [2]河北省中西医结合肝肾病证研究重点实验室,石家庄050091 [3]河北中医药大学中西医结合研究所,石家庄050091

出　　处：《中华中医药杂志》2023年第10期4891-4896,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：河北省中医药联合基金重点项目(No.H2022423351);河北省自然科学基金项目(No.H2021423006)。

摘　　要：目的:观察益气活血通络方调控孤核受体4A(NR4A1)/转化生长因子-β1(TGF-β1)负反馈环路对单侧输尿管梗阻(UUO)大鼠肾小管上皮间充质转化(EMT)的影响。方法:将48只雄性Wistar大鼠随机分为假手术组、UUO组、氯沙坦组、益气活血通络方组,每组12只。除假手术组外,其余各组大鼠予以UUO造模。14 d后取材行HE和Masson染色观察肾脏组织病理形态;免疫荧光检测α-平滑肌肌动蛋白(α-SMA)和E-钙黏附蛋白(E-cadherin)共定位,Western blot检测α-SMA、E-cadherin、NR4A1、TGF-β1的表达。体外采用TGF-β1刺激人肾小管上皮细胞(HK-2),予以过表达NR4A1及益气活血通络方含药血清干预,Western blot检测NR4A1、α-SMA、E-cadherin、TGF-β1及vimentin表达变化。结果:与假手术组比较,UUO组肾小管扩张、萎缩、炎症细胞浸润及胶原沉积增多;TGF-β1、α-SMA、vimentin表达显著升高(P<0.05),NR4A1、E-cadherin表达显著降低(P<0.05);与UUO组比较,各治疗组肾小管损伤程度明显改善,胶原沉积减少;NR4A1、E-cadherin表达显著上调(P<0.05),TGF-β1、α-SMA、vimentin表达显著降低(P<0.05)。TGF-β1刺激HK-2后,NR4A1、E-cadherin表达显著降低(P<0.05),α-SMA、vimentin、TGF-β1表达显著升高(P<0.05),过表达NR4A1后,E-cadherin表达显著升高(P<0.05),α-SMA、vimentin、TGF-β1表达显著降低(P<0.05),中药可增加NR4A1表达来降低α-SMA、vimentin、TGF-β1表达,增加E-cadherin的表达(P<0.05)。结论:益气活血通络方可通过调控NR4A1/TGF-β1负反馈环路抑制UUO大鼠EMT改善肾纤维化。Objective:To observe the effect of the NR4A1/TGF-β1 negative feedback loop on epithelial-mesenchymal transition(EMT)in rats with unilateral ureteral obstruction(UUO),which is regulated by Yiqi Huoxue Tongluo Recipe.Methods:Forty-eight male Wistar rats were randomly divided into sham group,UUO group,losartan group and Yiqi Huoxue Tongluo Recipe group,12 rats in each group.Except for the sham group,the unilateral ureter of each group was ligated to replicate the renal fibrosis model.Fourteen days after sampling,the renal function of each group was examined;the histomorphological changes of the kidney were observed by HE and Masson staining;Immunofluorescence double staining detection Co-localization of α-smooth muscle actin(α-SMA)and E-cadherin,the expression of α-SMA,E-calcadherin,nuclear receptor subfamily 4 group A member 1(NR4A1),transforming growth factor-β1(TGF-β1)were detected by Western blot.TGF-β1 was used to stimulate renal tubular epithelial cells(HK-2)in vitro,and NR4A1 and drug-containing serum were overexpressed to intervene,and changes in NR4A1,α-SMA,E-cadherin,TGF-β1 and vimentin expression were detected by Western blot.Results:Compared with the sham group,the UUO group showed increased renal tubular dilation,atrophy,infammatory cell infiltration,and collagen deposition,the expression of TGF-β1 and α-SMA was significantly increased(P<0.05).while the expression of NR4A1 and E-cadherin was significantly reduced(P<0.05).Compared with the UUO group,the degree of renal tubular injury was significantly improved and collagen deposition was reduced in each treatment group,the expression of NR4Al and E-cadherin significantly upregulated(P<0.05)and TGF-β1 and α-SMA was significantly reduced(P<0.05).After TGF-β1 stimulation of HK-2,the expression of NR4A1 and E-cadherin was significantly reduced(P<0.05),α-SMA,vimentin and TGF-β1 was significantly upregulated.While the expression of E-cadherin was significantly increased after overexpression of NR4A1(P<0.05),α-SMA,vimentin,TGF-β1 was significa

关 键 词：益气活血通络方 单侧输尿管梗阻 上皮间充质转化 肾纤维化 孤核受体4A1 转化生长因子-β1 

分 类 号：R285.5[医药卫生—中药学]