摘要：
This study was aimed to explore the renoprotective effects of Tang-Shen-Ping (TSP) on RhoA/ROCK signaling pathway in KKAy mice with diabetic kidney disease (DKD).A total of 60 female 10-week SPF degree KKAy mice,which were fed with KK special food for 10 weeks,were made into DKD model.Mice were randomly divided in the model group,irbesartan group,low-,medium-and high-dose TSP group (0.525 g· kg-1,1.05 g· kg-1,and 2.1 g· kg-1).Ten female C57BL/6J mice were used as the normal control group.Mice of each group were intragastrically administered with corresponding medicine,respectively,while mice of the control group and the model group were given deionized water of the equal volume.The body weight was measured and the 24-hour urine protein quantification was detected every 4 weeks.At the end of the 26th week,all mice were sacrificed and the biochemical indicators,such as fasting blood glucose (FBG),serum blood urea nitrogen (BUN),serum creatinine (Scr),and triglyceride (TG) were measured.HE staining,Mallory staining and PAS staining were used to observe the pathological morphology of kidney tissues.Immunohistochemistry (IHC) and in situ hybridization (ISH) were used in the detection of transforming growth factor-β1 (TGF-β1),Ras homolog gene family member A (RhoA),Rho-associated coiled-coil-containing protein kinase 1 (ROCK1),α-smooth muscle actin (α-SMA),E-Cadherin (E-Cad) mRNA and protein expression.The results showed that compared with the model group,there were significant differences on body weight,the ratio of kidney weight to body weight,and urinary protein in the middle-and high-dose TSP group (P ＜ 0.01);the renal pathological damage was obviously decreased;contents of FBG,BUN,Scr and TG decreased (P ＜ 0.01);mRNA and protein expression of E-Cadherin increased;mRNA and protein expression of TGF-β1,RhoA,ROCK1 and α-SMA decreased with significant difference in the middle-and high-dosc TSP group (P ＜ 0.01).It was concluded that the renoprotective effects and epithelial-mesenchymal transdifferentiation (EMT) of renal tubular epithelial cells of TSP on DKD KKAy mice may be related to the regulation of RhoA/ROCK signaling pathway.%目的:探讨糖肾平对糖尿病肾病(Diabetic Kidney Disease,DKD)KKAy小鼠肾保护作用及其对RhoA/ROCK信号通路的影响.方法:雌性10周龄SPF级KKAy小鼠60只,KK鼠料诱导10周建立DKD模型,随机分为模型组、厄贝沙坦组、糖肾平低(0.525 g·kg-1)、中(1.05 g·kg-1)、高(2.1 g·kg-1)剂量组,10只雌性C57BL/6J小鼠为正常组.各治疗组灌胃给药,正常组、模型组灌胃等体积去离子水,每4周称体质量并测24 h尿蛋白定量.第26周小鼠麻醉后摘眼球取血,称肾质量、测空腹血糖(Fasting Blood Glucose,FBG)、血尿素氮(Blood Urea Nitrogen,BUN)、血肌酐(Serum creatinine,Scr)和甘油三酯(Triglyceride,TG)含量;HE、Mallory和PAS染色观察肾组织病理;免疫组化、原位杂交测肾组织转化生长因子-β1(Transforming growth factor-β1,TGF-β1)、Ras同源基因家族成员A (Ras homolog gene familymember A,RhoA)、Rho相关卷曲蛋白激酶(Rho-assoeiatedcoiledcoi1-containing protein kinase 1,ROCK1)、α-平滑肌肌动蛋白(α-Smooth Muscle Actin,α-SMA)、E-钙黏素(E-Cadherin,E-Cad)mRNA及蛋白表达.结果:与模型组相比,各治疗组体质量、肾质量/体质量、尿蛋白降低,糖肾平中、高剂量组有显著性差异(P＜0.01);肾病理损害明显减轻,FBG、BUN、Scr、TG含量降低(P＜0.01),E-Cadherin mRNA及蛋白表达增加,TGF-β1、RhoA、ROCK1和α-SMA mRNA和蛋白表达减少,糖肾平中、高剂量组差异显著(P＜0.01).结论:糖肾平对DKD KKAy小鼠肾的保护作用及抑制肾小管上皮细胞转分化的作用,可能与调节RhoA/ROCK信号通路有关.
