Page 23 - 南京医科大学学报自然科学版

第43卷第9期 南京医科大学学报（自然科学版）
2023年9月 Journal of Nanjing Medical University（Natural Sciences） ·1201 ·

·基础研究·

达格列净通过Rffl 抑制STAT1/TGF⁃β1信号通路改善糖尿病肾

病肾小管上皮细胞EMT和纤维化

徐 洋，王 敏，张恒璐，周 莉，陆卫平 *

南京医科大学附属淮安第一医院内分泌科，江苏 淮安 223300

［摘 要］ 目的：探讨达格列净对糖尿病肾病肾小管上皮细胞的上皮细胞⁃间充质转化和纤维化的影响及分子机制。方法：体
外培养人肾小管上皮细胞HK⁃2，分为对照组、高糖组、低剂量达格列净+高糖组和高剂量达格列净+高糖组。用Western blot与
RT⁃PCR 分别检测 E3 泛素连接酶 Rffl 的表达水平。Western blot 检测各组上皮细胞钙黏蛋白（epithelial cadherin，E⁃cadherin）、
α⁃平滑肌肌动蛋白（α⁃smooth muscle actin，α⁃SMA）、纤连蛋白（Fibronectin）、转化生长因子⁃β1（transforming growth factor β1，
TGF⁃β1）、信号转导与转录激活因子1（signal transducer and activator of transcription 1，STAT1）的表达水平。在HK⁃2细胞中过
表达Rffl后，Western blot 检测E⁃cadherin、α⁃SMA、Fibronectin、TGF⁃β1、STAT1的表达水平。结果：高糖组中Rffl的表达水平显
著低于对照组，加入达格列净后Rffl表达升高；与对照组相比，高糖组细胞中E⁃cadherin表达水平降低，Fibronectin、α⁃SMA表达
水平升高，过表达Rffl后E⁃cadherin表达水平升高，Fibronectin、α⁃SMA表达水平降低；与高糖组相比，低剂量达格列净+高糖组
和高剂量达格列净+高糖组中E⁃cadherin表达水平均升高，Fibronectin、α⁃SMA表达水平均降低，且呈一定的剂量依赖性；与对
照组相比，高糖组细胞中STAT1、TGF⁃β1的表达水平升高，而在过表达Rffl或达格列净作用后则明显降低。结论：达格列净通
过上调 Rffl的表达抑制STAT1/TGF⁃β1信号通路，改善糖尿病肾病肾小管上皮细胞的上皮细胞⁃间充质转化和纤维化。
［关键词］ 达格列净；糖尿病肾病；Rffl；TGF⁃β1；EMT；纤维化
［中图分类号］ R587.2 ［文献标志码］ A ［文章编号］ 1007⁃4368（2023）09⁃1201⁃07
doi：10.7655/NYDXBNS20230903

Dapagliflozin improves renal tubular epithelial EMT and fibrosis in diabetic nephropathy
by inhibiting STAT1/TGF⁃β1 signaling pathway via Rffl

XU Yang，WANG Min，ZHANG Henglu，ZHOU Li，LU Weiping *
Department of Endocrinology and Metabolism，the Affiliated Huai’an No.1 People’s Hospital of Nanjing Medical
University，Huai’an 223300，China

［Abstract］ Objective：The current study aims to investigate the effect and molecular mechanism of dapagliflozin on epithelial ⁃
mesenchymal transformation and fibrosis of renal tubular epithelial cells in diabetic kidney disease. Methods：Human renal tubular
epithelial cells HK⁃2 were cultured in vitro and divided into control group，high glucose group，low dose dapagliflozin + high glucose
group and high dose dapagliflozin+high glucose group. The expression levels of Rffl were detected by Western blot and RT⁃PCR，
respectively. The expression levels of epithelial cadherin（E⁃cadherin），α⁃smooth muscle actin（α⁃SMA），Fibronectin，transforming
growth factor β1（TGF⁃β1）and signal transducer and activator of transcription 1（STAT1）were detected by Western blot. After Rffl
was overexpressed in HK⁃2 cells，the expression levels of E⁃cadherin，α⁃SMA，Fibronectin，TGF⁃β1 and STAT1 were detected by
Western blot. Results：The expression level of Rffl was significantly lower in the high glucose group than in the control group，and the
expression of Rffl was increased after adding dapagliflozin. Compared with the control group，the expression level of E⁃Cadherin in the
high glucose group was decreased，while the expression level of Fibronectin and α⁃SMA was increased. After Rffl was overexpressed in
cells，the expression level of E ⁃ Cadherin was increased，while the expression levels of Fibronectin and α ⁃ SMA were decreased.
Compared with high glucose group，the expression levels of E⁃Cadherin in low⁃dose dapagliflozin+high glucose group and high⁃dose

［基金项目］ 国家自然科学基金（81800644）；淮安市“533英才”工程科研项目资助计划（HAA201744）
∗
通信作者（Corresponding author），E⁃mail：hyhalwp@sina.com