miRNA-139调控Wnt/β-catenin信号通路探讨建中补肾消癥汤抗肾间质纤维化机制  

Investigation on Mechanism of Jianzhong Bushen Xiaozheng Decoction Against Renal Interstitial Fibrosis in Regulating Wnt/β-catenin Signaling Pathway by MiRNA-139

作　　者：刘武 丁宁 姜赫 李子腾[2] 张诏 LIU Wu;DING Ning;JIANG He;LI Zi-teng;ZHANG Zhao(College of Traditional Chinese Medicine(TCM),Shandong University of TCM,Ji'nan 250355,China;College of Acupuncture and Massage,Shandong University of TCM,Ji'nan 250355,China)

机构地区：[1]山东中医药大学中医学院,济南250355 [2]山东中医药大学针灸推拿学院,济南250355

出　　处：《中国实验方剂学杂志》2021年第4期47-55,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金面上项目(81273635);山东省中医药科技发展计划项目(2019-0043);山东省自然科学基金面上项目(ZR2020MH344)。

摘　　要：目的:探讨建中补肾消癥汤在调控miRNA-139作用于Wnt/β-连环蛋白(β-catenin)信号通路以调节肾间质纤维化的作用机制。方法:将120只小鼠随机分为假手术组,UUO组,建中补肾消癥汤低、中、高剂量组,尿毒清组,采用单侧输尿管梗阻(UUO)动物模型。造模术后3 d开始灌胃给药,假手术组,UUO组每天予以等量蒸馏水灌胃,建中补肾消癥汤低、中、高剂量组分别按6,12,24 g·kg^(-1)·d^(-1)给予建中补肾消癥汤水溶液灌胃,尿素清组予6.2 g·kg^(-1)·d^(-1)给予尿毒清颗粒水溶液灌胃,观察14,21,28 d后小鼠的形态变化,观察小鼠一般体征及梗阻侧肾间质纤维化指数,采用苏木素-伊红(HE)染色及马松(Masson)染色观察肾组织病理形态变化,实时荧光定量聚合酶链式反应(Real-time PCR)检测梗阻侧肾组织中miRNA-139的表达量,蛋白免疫印迹法(Western blot)检测梗阻侧肾组织中β-catenin,纤溶酶原激活物抑制剂-1(PAI-1)的蛋白表达,免疫组化检测梗阻侧肾组织中基质金属蛋白酶-7(MMP-7)蛋白的表达。结果:14,21,28 d后,与假手术组比较,UUO组小鼠β-catenin,PAI-1蛋白表达升高(P<0.05),miRNA-139,MMP-7蛋白表达降低(P<0.05),与UUO组比较,经尿毒清组及各中药组治疗后小鼠β-catenin,PAI-1蛋白表达明显降低(P<0.05),miRNA-139,MMP-7蛋白表达明显升高(P<0.05),且建中补肾消癥汤高剂量组的疗效优于其他剂量及尿毒清组(P<0.05)。结论:建中补肾消癥汤可能通过对miRNA-139的干预作用,调控Wnt/β-catenin通路起到抗肾纤维作用,达到保护肾功能,延缓慢性肾脏病(CKD)进展的效果。Objective: To investigate the mechanism of Jianzhong Bushen Xiaozheng decoction in regulating the effect of miRNA139 on Wnt/β-catenin signaling pathway for renal interstitial fibrosis. Method:The 120 mice were randomly divided into sham operation group,unilateral ureteral obstruction(UUO)group,Jianzhong Bushen Xiaozheng decoction low,middle,high dose group,and Niaoduqing group. The UUO animal model was established to observe the morphological changes in mice. Intragastic administration was started from day 3 after modeling. The sham operation group and UUO group received the same amount of distilled water every day. The low,medium and high-dose groups received Jianzhong Bushen Xiaozheng decoction solution at6,12,24 g·kg^(-1)·d^(-1) respectively. The Niaoduqing group received 6.2 g·kg^(-1)·d^(-1) Niaoduqing granule solution.After 14 d and 21 d,28 d,the morphological changes,general signs and renal interstitial fibrosis index of the obstructed side were observed,hematoxylin-eosin(HE)staining and Masson staining were used to observe the pathological changes of renal tissue. Real-time fluorescent quantitative polymerase chain reaction(Real-time PCR)method was used to detect the miRNA-139 expression in renal tissue volume,Western blot was used to detect expression of beta serial proteins(β-catenin)and fibrinolytic enzyme activators inhibitor-1(PAI-1)in renal tissues,and immunohistochemical assay was used for detection of matrix metalloproteinases-7(MMP-7)protein expression at the obstruction side. Result:After 14,21 and 28 days,the expression levels of β-catenin and PAI-1 in UUO group were higher than those in sham operation group(P<0.05),while the expression levels of miRNA139 and MMP-7 protein were lower than those of sham operation group(P<0.05). The expression levels of β-catenin and PAI-1 proteins in mice after treatment in Niaoduqing group and the traditional Chinese medicine groups were lower than those in the UUO group(P<0.05),the expression of miRNA139 and MMP-7 proteins increased(P<0.05),and the effi

关 键 词：miRNA-139 Wnt/β-连环蛋白(β-catenin)信号通路 肾间质纤维化 建中补肾消癥汤 实验研究 

分 类 号：R2-0[医药卫生—中医学] R22[医药卫生—中医基础理论]