Didang Decoction-medicated serum enhances autophagy in high glucose-induced rat glomerular endothelial cells via the PI3K/Akt/mTOR signaling pathway
10.12122/j.issn.1673-4254.2025.03.03
- VernacularTitle:抵当汤含药血清通过PI3K/Akt/mTOR信号通路增强高糖诱导的大鼠肾小球内皮细胞自噬
- Author:
Yanyan DONG
1
;
Kejing ZHANG
;
Jun CHU
;
Quangen CHU
Author Information
1. 安徽中医药大学中医学院,安徽 合肥 230012;安徽中医药大学新安医学教育部重点实验室,安徽 合肥 230038
- Publication Type:Journal Article
- Keywords:
diabetic nephropathy;
Didang decoction;
autophagy;
PI3K/Akt/mTOR signaling pathway;
glomerular endothelial cells
- From:
Journal of Southern Medical University
2025;45(3):461-469
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of Didang Decoction-medicated serum on autophagy in high glucose(HG)-induced rat glomerular endothelial cells(RGECs)and explore the pathway that mediates its effect.Methods Primary RGECs were isolated and cultured using sequential sieving combined with collagenase digestion,followed by identification using immunofluorescence assay for factor Ⅷ.High glucose medium was used to induce RGECs to simulate a diabetic environment,and the effects of Didang Decoction-medicated serum and 3-MA(an autophagy inhibitor),either alone or in combination,on autophagy of HG-exposed cells were evaluated by observing autophagic vacuoles using monodansylcadaverine(MDC)staining.RT-qPCR and Western blotting were employed to measure mRNA and protein expression levels of Beclin-1,p62,LC3B,p-PI3K,p-Akt,and p-mTOR.Results Compared with the control cells,the HG-exposed RGECs showed significantly reduced autophagic fluorescence intensity,decreased Beclin-1 mRNA expression,increased p62 mRNA expression,downregulated Beclin-1 protein and LC3-Ⅱ/Ⅰ ratio,and upregulated p62,p-PI3K,p-Akt,and p-mTOR protein levels.Didang Decoction-medicated serum significantly enhanced autophagic fluorescence intensity in HG-exposed cells,increased Beclin-1 mRNA expression,decreased p62 mRNA expression,upregulated Beclin-1 protein,and downregulated p62,p-PI3K,p-Akt,and p-mTOR protein levels.Conclusion Didang Decoction-medicated serum enhances autophagy in HG-exposed RGECs by regulating the PI3K/Akt/mTOR signaling pathway,which sheds light on a new therapeutic strategy for diabetic nephropathy.
