海南医科大学学报2025，Vol.31Issue(7)：481-489,9.DOI:10.13210/j.cnki.jhmu.20250210.001
益智仁抑制PFKFB3介导的糖酵解对糖尿病肾病上皮间充质转化的影响
Effect of inhibition of PFKFB3-mediated glycolysis on epithelial mesenchymal transition in diabetic kidney disease by Alpiniae Oxyphyllae Fructus
摘要
Abstract
Objective:To investigate the potential therapeutic effect of Alpiniae Oxyphyllae Fructus on epithelial mesenchymal transition in diabetic kidney disease using in vivo and in vitro models and to study its mechanism of action.Methods:In vivo exper-iments:db/m mice were used as a blank group,and db/db mice were divided into the model group,the positive drug control group(the Empagliflozin group),and the low,medium,and high dose groups of Alpiniae Oxyphyllae Fructus.The pathological changes of renal tissues were observed by H&E staining,and the mRNA and protein expression levels of PFKFB3,E-cadherin,Col3α1 and Vimentin in renal tissues were detected by RT-qPCR and Western blot.In vitro experiments:HK-2 cells were divided into the blank group,the model group,the different dose groups(low,medium,and high dose groups)of Alpiniae Oxyphyllae Fructus-containing serum,and the positive control group of Empagliflozin.After 24 h of drug intervention,the migration ability of cells in each group was firstly assessed by scratch assay,and the optimal drug concentration was determined accordingly.Subse-quently,the mRNA and protein expression levels of PFKFB3,E-cadherin,Col3α1 and Vimentin in HK-2 cells were detected by RT-qPCR and Western blot;the expression of the key protein Col3α1 in EMT was further verified by immunofluorescence.Re-sults:In vivo experiments:H&E staining revealed that,in comparison to the blank group,the model group exhibited an increase in glomerular volume,thickening of the basement membrane,swelling of tubular epithelial cells,and narrowing of the tubular lu-men.After intervention with Alpiniae Oxyphyllae Fructus and Empagliflozin,renal pathological changes were observed to be re-duced to varying degrees.The model group showed a significant downregulation of E-cadherin mRNA and protein expression(P<0.05)compared to the blank group,while the expression levels of PFKFB3,Col3α1,and Vimentin were significantly upregulated in each administered group(P<0.05).In contrast to the model group,the administered groups demonstrated a significant upregu-lation of E-cadherin expression(P<0.05)and a significant downregulation of PFKFB3,Col3α1,and Vimentin expression(P<0.05).In vitro experiments:The model group's cell migration ability was significantly enhanced(P<0.05)compared to the blank group.The migration ability of HK-2 cells was reduced in all serum-containing groups and the Empagliflozin group(P<0.05),with the 10%serum-containing group showing the most pronounced reduction.The model group displayed a significant downregu-lation of E-cadherin mRNA and protein expression(P<0.05)and an upregulation of PFKFB3,Col3α1,and Vimentin mRNA and protein expression(P<0.05)compared to the blank group.In the drug-administered groups,E-cadherin expression was signif-icantly upregulated(P<0.05),while the expression of PFKFB3,Col3α1,and Vimentin was significantly downregulated(P<0.05)when compared to the model group.Additionally,the fluorescence expression level of Col3α1 was elevated in the model group compared to the blank group,and this level decreased following drug treatment.Conclusion:Alpiniae Oxyphyllae Fructus improves epithelial mesenchymal transition in diabetic nephropathy by inhibiting PFKFB3-mediated abnormal glycolysis.关键词
益智仁/糖尿病肾病/糖酵解/上皮间充质转化Key words
Alpiniae Oxyphyllae Fructus/Diabetic kidney disease/Glycolysis/Epithelial-mesenchymal transition分类
中医学引用本文复制引用
李小燕,袁勇,王晶,马天鹏,李丽丹,颜子杰,肖曼,谢毅强..益智仁抑制PFKFB3介导的糖酵解对糖尿病肾病上皮间充质转化的影响[J].海南医科大学学报,2025,31(7):481-489,9.基金项目
This study was supported by the National Natural Science Foundation of China-General Project(82174334) （82174334）
Key Experiment on Traditional Chinese Medicine Regulation of Metabolic Diseases in Sichuan Province,2022 Open Fund(SZKF202203) 国家自然科学基金-面上项目(82174334) （SZKF202203）
代谢性疾病中医药调控四川省重点实验2022年开放基金(SZKF202203) （SZKF202203）
