Effect and mechanism of extracts from oxytropis falcata on improv-ing oxidative damage of podocytes induced by high glucose via regu-lating PI3K/AKT/Nrf2 pathway
10.12092/j.issn.1009-2501.2025.10.008
- VernacularTitle:镰形棘豆提取物调节PI3K/AKT/Nrf2通路改善高糖诱导足细胞氧化损伤的作用及机制研究
- Author:
Menghan REN
1
;
Jianli YAO
;
Lixia YANG
;
Chang SHU
Author Information
1. 甘肃中医药大学,兰州 730000,甘肃
- Publication Type:Journal Article
- Keywords:
oxytropis falcata extract;
diabetic kid-ney disease;
podocyte;
oxidative stress;
PI3K/AKT/Nrf2
- From:
Chinese Journal of Clinical Pharmacology and Therapeutics
2025;30(10):1361-1373
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To investigate the effect of drug-containing serum of oxytropis falcata extract on oxi-dative stress injury of podocyte induced by high glucose through PI3K/AKT/Nrf2 pathway.METH-ODS:The rat renal podocyte was cultured in vitro,and the concentration and time of D-glucose mod-elling and the optimal concentration and time of administration in the drug-containing serum of oxy-tropis falcata extract were screened.The cells were divided into normal group,high glucose group,high glucose+blank serum group,oxytropis falcata group,inhibitor group,oxytropis falcata+inhibitor group.Rhodamine staining and electron microsco-py were used to observe the morphological and pathological changes of podocyte,flow cytometry was used to detect apoptosis rate,and cell migra-tion ability was detected by scratch test.Immuno-fluorescence and fluorescence probe were used to detect the fluorescence expression of p-AKT and ROS level of cells,ELISA was used to detect the con-tent of MDA,NO,SOD and T-AOC in the superna-tant of cells,and Western Blot was used to detect the protein expression of p-PI3K/PI3K,p-AKT/AKT and Nrf2 in cells.The mRNA expressions of Nrf2,HO-1,GCLM and SOD were detected by RT-qPCR.RESULTS:It was selected that 45 mmol/LD-glucose induction for 48 hours was the best modeling con-dition,and the 1-fold dilution of the medicated se-rum of oxytropis falcata extract for 48 hours was the best concentration and intervention time.Com-pared with the normal group,the foot processes of the high glucose group were widely fused and ad-hered to each other,the apoptosis rate,migration ability and intracellular ROS level were significantly increased(P＜0.01),the fluorescence expression of p-AKT was markedly decreased(P＜0.01),the con-tents of MDA and NO were dramatically enhanced,and the contents of SOD and T-AOC were signifi-cantly downregulated(P＜0.01).The protein expres-sion of p-PI3K/PI3K,p-AKT/AKT,Nrf2 and the mRNA expression of Nrf2,HO-1,GCLM and SOD markedly declined(P＜0.01).Compared with high glucose group,foot process fusion and adhesion of podo-cytes in oxytropis falcata group and oxytropis falca-ta+inhibitor group were improved in varying de-grees,apoptosis rate,migration ability and intracel-lular ROS level significantly declined(P＜0.05,P＜0.01),p-AKT fluorescence expression increased(P＜0.01),NO content decreased,T-AOC level elevated(P＜0.01);the content of MDA decreased and the activity of SOD notably rose(P＜0.01),the protein expression of p-PI3K/PI3K,p-AKT/AKT,Nrf2 and the mRNA expression of Nrf2,HO-1,GCLM and SOD markedly increased in oxytropis falcata group(P＜0.05,P＜0.01);the level of ROS in podocytes im-proved(P＜0.01),the fluorescence expression of p-AKT decreased(P＜0.05),the content of NO upregu-lated,the content of T-AOC downregulated,and the expression of p-PI3K/PI3K,p-AKT/AKT,Nrf2 pro-tein and HO-1,GCLM,SOD mRNA decreased in in-hibitor group(P＜0.05,P＜0.01).Compared with oxy-tropis falcata group,the apoptosis rate,migration ability and intracellular ROS level of podocytes in oxytropis falcata+inhibitor group markedly in-creased(P＜0.05,P＜0.01),p-AKT fluorescence ex-pression declined(P＜0.01),MDA and NO content increased,SOD and T-AOC content decreased(P＜0.05,P＜0.01),p-PI3K/PI3K,p-AKT/AKT,Nrf2 protein and Nrf2,HO-1,GCLM,SOD mRNA expression all dramatically downregulated(P＜0.05,P＜0.01).CON-CLUSION:Oxytropis falcata extract may protect podocytes by activating the PI3K/AKT/Nrf2 signal-ing pathway,thereby improving the morphological,structural and functional changes of podocytes in-duced by high glucose and inhibiting oxidative stress response.
