Jiang Hong, Gao Mengya, Liu Jiaqi, Yang Lijuan, Liu Lei
Department of Nephrology, The First Affiliated Hospital of Bengbu Medical University, Bengbu, Anhui, People's Republic of China.
Department of Physiology, Bengbu Medical University, Bengbu, Anhui, People's Republic of China.
Diabetes Metab Syndr Obes. 2025 Jun 10;18:1903-1914. doi: 10.2147/DMSO.S515034. eCollection 2025.
Investigating the effects and mechanisms of dapagliflozin on pyroptosis of renal tubular epithelial cells under high-glucose conditions through the regulation of the Caspase-3/GSDME signaling pathway, providing experimental evidence for the clinical treatment of diabetic kidney disease.
Human renal tubular epithelial cells (HK-2) were cultured in vitro and divided into control group (5mmol/L D-glucose), high-glucose group (30mmol/L D-glucose), dagliflozin group (2.5μmol/L dagliflozin), and monoinhibitor group (20μmol/L caspase-1 inhibitor), dual inhibitor group (20μmol/L caspase-1 inhibitor + 50μmol/L caspase-3 inhibitor), and SiRNA transfection group. All groups were intervened for 48h. The cell viability was detected by cell counting kit-8 and the glucose and dagliflozin concentrations of the intervention were determined. Caspase-1, caspase-3, GSDMD, GSDME, GSDME-N, caspase-8, NF-κB were detected by Western blot. Detection of cellular pyroptosis in each group by flow cytometry.
Compared with the control group, the D-glucose group showed decreased cell viability, increased cell pyroptosis, and increased levels of caspase-1, caspase-3, GSDMD, GSDME, GSDME-N, caspase-8, NF-κB, and other related proteins (P<0.05). Compared with the D-glucose group, the rate of cellular pyroptosis and the levels of caspase-1, caspase-3, GSDMD, GSDME, GSDME-N and other related proteins were decreased in the dagliflozin group and the dual inhibitor group (<0.05). Compared with the transfected control group, the cellular pyroptosis rate and the levels of caspase-1, caspase-3, GSDMD, GSDME, GSDME-N, and other related proteins were then further reduced in the transfected group targeting SGLT2 knockdown (<0.05).
In the proximal tubular cells of diabetic kidney disease, dagliflozin inhibited high glucose-induced pyroptosis of human HK-2, and its mechanism of action may be related to the inhibition of caspase 3/GSDME pathway signaling.
通过调控半胱天冬酶-3/ Gasdermin-E（Caspase-3/GSDME）信号通路，研究达格列净对高糖条件下肾小管上皮细胞焦亡的影响及机制，为糖尿病肾病的临床治疗提供实验依据。
体外培养人肾小管上皮细胞（HK-2），分为对照组（5mmol/L D-葡萄糖）、高糖组（30mmol/L D-葡萄糖）、达格列净组（2.5μmol/L达格列净）、单抑制剂组（20μmol/L半胱天冬酶-1抑制剂）、双抑制剂组（20μmol/L半胱天冬酶-1抑制剂+50μmol/L半胱天冬酶-3抑制剂）和小干扰RNA（SiRNA）转染组。所有组干预48小时。采用细胞计数试剂盒-8检测细胞活力，并测定干预的葡萄糖和达格列净浓度。通过蛋白质免疫印迹法检测半胱天冬酶-1、半胱天冬酶-3、Gasdermin-D（GSDMD）、Gasdermin-E（GSDME）、裂解的Gasdermin-E（GSDME-N）、半胱天冬酶-8、核因子-κB（NF-κB）。通过流式细胞术检测各组细胞焦亡情况。
与对照组相比，D-葡萄糖组细胞活力降低，细胞焦亡增加，半胱天冬酶-1、半胱天冬酶-3、GSDMD、GSDME、GSDME-N、半胱天冬酶-8、NF-κB等相关蛋白水平升高（P<0.05）。与D-葡萄糖组相比，达格列净组和双抑制剂组细胞焦亡率及半胱天冬酶-1、半胱天冬酶-3、GSDMD、GSDME、GSDME-N等相关蛋白水平降低（P<0.05）。与转染对照组相比，靶向沉默钠-葡萄糖协同转运蛋白2（SGLT2）的转染组细胞焦亡率及半胱天冬酶-1、半胱天冬酶-3、GSDMD、GSDME、GSDME-N等相关蛋白水平进一步降低（P<0.05）。
在糖尿病肾病近端肾小管细胞中，达格列净抑制高糖诱导的人HK-2细胞焦亡，其作用机制可能与抑制半胱天冬酶3/GSDME信号通路有关。
