基于AP-1/TGF-β1/Smad3信号通路探讨左归降糖益肾方对糖尿病肾病小鼠的肾脏保护作用*

作者：陈莉莉1，2，胡淑娟1，李旭华2，彭 瑶1，2，周彤艺1，喻 嵘1，彭亚军2

单位：1.湖南中医药大学，湖南 长沙 410208； 2.湖南中医药大学第一附属医院，湖南 长沙 410007

引用：引用：陈莉莉，胡淑娟，李旭华，彭瑶，周彤艺，喻嵘，彭亚军.基于AP-1/TGF-β1/Smad3信号通路探讨左归降糖益肾方对糖尿病肾病小鼠的肾脏保护作用[J].中医药导报,2026,32(5):41-48.

DOI：10.13862/j.cn43-1446/r.2026.05.007

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摘要：

目的：探讨左归降糖益肾方对糖尿病肾病小鼠的肾脏保护作用及可能的作用机制。方法：50只db/db小鼠，普通饲料喂养8周后，建立糖尿病肾病小鼠模型。模型小鼠随机分为模型组、左归降糖益肾方低剂量组、左归降糖益肾方中剂量组、左归降糖益肾方高剂量组及达格列净组，每组10只。10只db/m小鼠设为正常组。左归降糖益肾方低、中、高剂量组分别予左归降糖益肾方药液（6.4、12.8、25.6 g/kg）灌胃，达格列净组小鼠予达格列净溶液（1.3 mg/kg）灌胃，干预8周。正常组和模型组小鼠予等体积生理盐水灌胃。给药期间，观察小鼠空腹血糖（FBG）、尿微量白蛋白/肌酐比值（UACR）变化。干预8周后，检测白蛋白（ALB）、丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、血肌酐（Scr）、尿素氮（BUN）、尿酸（UA）；苏木精-伊红（HE）染色、过碘酸希夫染色（PAS）、马松（Masson）染色及透射电镜（TEM）观察小鼠肾组织病理改变；实时荧光定量聚合酶链式反应（Real-time PCR）检测肾组织紧密连接蛋白1（ZO-1） mRNA、激活蛋白1（AP-1） mRNA、转化生长因子-β1（TGF-β1） mRNA、Smad家族成员3（Smad3） mRNA表达水平；蛋白质印迹法（Western blotting）检测肾组织ZO-1、AP-1、TGF-β1、Smad3蛋白表达水平。结果：第2、4、6、8周，模型组小鼠FBG高于正常组（P<0.05）；第6、8周，左归降糖益肾方低、中、高剂量组及达格列净组小鼠FBG低于模型组（P<0.05）。第4、8周，模型组小鼠UACR水平高于正常组（P<0.05）；第4周，左归降糖益肾方低、高剂量组小鼠UACR水平低于模型组（P＜0.05）；第8周，左归降糖益肾方低、中、高剂量组及达格列净组小鼠UACR水平低于模型组（P<0.05）。模型组小鼠血清Scr、BUN、UA、ALB、ALT、AST与正常组比较，差异均无统计学意义（P＞0.05）；左归降糖益肾方低、中、高剂量组及达格列净组小鼠血清Scr、BUN、UA、ALB、ALT、AST与模型组比较，差异均无统计学意义（P＞0.05）。正常组小鼠肾小球、肾小管结构完整；与正常组比较，模型组小鼠肾小球足突融合，系膜细胞和基质增生，基底膜增厚，间质纤维增生；与模型组比较，左归降糖益肾方低、中、高剂量组及达格列净组小鼠肾小球及其内皮细胞、足细胞病变改善。模型组小鼠肾组织糖原阳性面积占比和胶原纤维阳性面积占比大于正常组（P＜0.05）；左归降糖益肾方低、中、高剂量组及达格列净组小鼠肾组织糖原阳性面积占比和胶原纤维阳性面积占比小于模型组（P＜0.05）。模型组小鼠肾组织ZO-1 mRNA、Smad3 mRNA相对表达量高于正常组（P<0.05）；左归降糖益肾方低、中、高剂量组和达格列净组小鼠肾组织ZO-1 mRNA、Smad3 mRNA相对表达量低于模型组（P<0.05）。模型组小鼠肾组织ZO-1、AP-1、Smad3蛋白相对表达量高于正常组（P<0.05）；左归降糖益肾方低、中、高剂量组及达格列净组小鼠肾组织Smad3蛋白相对表达量低于模型组（P<0.05）。结论：左归降糖益肾方可能通过调控AP-1/TGF-β1/Smad3信号通路，改善糖代谢，保护足细胞，从而延缓糖尿病肾病进展。

关键词：糖尿病肾病；左归降糖益肾方；2型糖尿病；db/db小鼠；AP-1/TGF-β1/Smad3信号通路

Abstract：

Objective: To investigate the renal protective effect and possible mechanism of Zuogui Jiangtang Yishen formula (ZGJTYSF) in mice with diabetic kidney disease. Methods: Totally 50 db/db mice were fed with normal diet for 8 weeks to establish a diabetic kidney disease model. The model mice were randomly divided into a model group, a low-dose ZGJTYSF group, a medium-dose ZGJTYSF group, a high-dose ZGJTYSF group, and a dapagliflozin group, with 10 mice in each group. A total of 10 db/m mice served as the normal group. The low-dose ZGJTYSF group, medium-dose ZGJTYSF group, and high-dose ZGJTYSF group were administered the corresponding drugs (6.4,12.8,25.6 g/kg) by gavage respectively, while the dapagliflozin group was administered with dapagliflozin solution (1.3 mg/kg) by gavage. All groups received continuous administration for 8 weeks. The normal group and model group were administered with an equal volume of normal saline by gavage. During the administration period, changes in fasting blood glucose (FBG) and urinary albumin-to-creatinine ratio (UACR) were observed. After 8 weeks of drug administration, albumin (ALB), alanine aminotransferase (ALT), aspartate aminotransferase (AST), serum creatinine (Scr), blood urea nitrogen (BUN), and uric acid (UA) were measured. Hematoxylin-eosin (HE), periodic acid-Schiff (PAS), Masson staining, and transmission electron microscopy (TEM) were used to observe the pathological changes of renal tissue. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect the expressions of Zonula occludens-1 (ZO-1) mRNA, Activator protein 1 (AP-1) mRNA, Transforming growth factor-β1 (TGF-β1) mRNA, and Smad family member 3 (Smad3) mRNA in renal tissue. Western blotting was used to detect the expression of ZO-1, AP-1, TGF-β1, and Smad3 proteins. Results: FBG in the model group was significantly higher than that in the normal group at weeks 2nd, 4th, 6th, and 8th (P<0.05). FBG in the low-dose ZGJTYSF group, medium-dose ZGJTYSF group, high-dose ZGJTYSF group, and dapagliflozin group were significantly lower than that in the model group at weeks 6th and 8th (P<0.05). UACR in the model group were significantly higher than that in the normal group at weeks 4th and 8th (P<0.05). UACR in the low-dose ZGJTYSF group and high-dose ZGJTYSF group was significantly lower than that in the model group at week 4th (P<0.05). UACR in the low-dose ZGJTYSF group, medium-dose ZGJTYSF group, high-dose ZGJTYSF group, and dapagliflozin group was significantly lower than that in the model group at week 8th (P<0.05). There were no significant differences in Scr, BUN, UA, ALB, ALT and AST between the model group and the normal group (P>0.05). Similarly, no statistically significant differences were observed in these indicators among the low-dose ZGJTYSF group, medium-dose ZGJTYSF group, high-dose ZGJTYSF group, and dapagliflozin group compared with the model group (P>0.05). The glomeruli and tubules of mice were intact in the normal group. Compared with the normal group, the model group exhibited podocyte foot process fusion, mesangial cells proliferation, mesangial matrix expansion, glomerular basement membrane thickening, and interstitial fibrosis. Compared with the model group, the pathological lesions of glomeruli as well as endothelial cells and podocytes were improved in the low-dose ZGJTYSF group, medium-dose ZGJTYSF group, high-dose ZGJTYSF group, and dapagliflozin group. The positive areas of glycogen and collagen fibers in the model group were significantly larger than those in the normal group (P<0.05). The positive areas of glycogen and collagen fibers in the low-dose ZGJTYSF group, medium-dose ZGJTYSF group, high-dose ZGJTYSF group, and dapagliflozin group were significantly smaller than those in the model group (P<0.05). Compared with the normal group, the relative expression levels of ZO-1 mRNA and Smad3 mRNA in renal tissue were significantly higher in the model group (P<0.05). The relative expression levels of ZO-1 mRNA and Smad3 mRNA in renal tissue were significantly lower in the low-dose ZGJTYSF group, medium-dose ZGJTYSF group, high-dose ZGJTYSF group, and dapagliflozin group than those in the model group (P<0.05). Compared with the normal group, the relative expression levels of ZO-1, AP-1 and Smad3 proteins in renal tissue were significantly higher in the model group (P<0.05). The relative expression level of Smad3 protein in renal tissue in the low-dose ZGJTYSF group, medium-dose ZGJTYSF group, high-dose ZGJTYSF group, and dapagliflozin group was significantly lower than that in the model group (P<0.05). Conclusion: Zuogui Jiangtang Yishen formula may delay the progression of diabetic kidney disease by regulating the AP-1/TGF-β1/Smad3 signaling pathway, improving glucose metabolism, and protecting podocytes.

Key words：diabetic kidney disease; Zuogui Jiangtang Yishen formula; type 2 diabetes; db/db mouse; AP-1/TGF-β1/Smad3 signaling pathway

发布时间：2026-05-23

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