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
- Author:
Yanyan DONG
1
;
Kejing ZHANG
1
;
Jun CHU
1
;
Quangen CHU
1
Author Information
1. School of Traditional Chinese Medicine, Anhui University of Chinese Medicine, Hefei 230012, China.
- Publication Type:Journal Article
- Keywords:
Didang decoction;
PI3K/Akt/mTOR signaling pathway;
autophagy;
diabetic nephropathy;
glomerular endothelial cells
- MeSH:
Animals;
Autophagy/drug effects*;
Signal Transduction/drug effects*;
Rats;
TOR Serine-Threonine Kinases/metabolism*;
Drugs, Chinese Herbal/pharmacology*;
Proto-Oncogene Proteins c-akt/metabolism*;
Endothelial Cells/metabolism*;
Phosphatidylinositol 3-Kinases/metabolism*;
Glucose;
Cells, Cultured;
Kidney Glomerulus/cytology*;
Rats, Sprague-Dawley
- From:
Journal of Southern Medical University
2025;45(3):461-469
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVES: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.
CONCLUSIONS: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.
