文献类型：期刊文献

中文题名：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

英文题名：Effect and mechanism of extracts from oxytropis falcata on improving oxidative damage of podocytes induced by high glucose via regulating PI3K/AKT/Nrf2 pathway

作者：任梦涵[1];姚建莉[1];杨丽霞[2];舒畅[1]

第一作者：任梦涵

机构：[1]甘肃中医药大学,甘肃兰州730000;[2]绍兴市人民医院,浙江绍兴312000

第一机构：甘肃中医药大学

年份：2025

卷号：30

期号：10

起止页码：1361

中文期刊名：中国临床药理学与治疗学

外文期刊名：Chinese Journal of Clinical Pharmacology and Therapeutics

收录：;北大核心:【北大核心2023】；

基金：兰州市科技计划项目(2021-1-94);甘肃省中医药管理局项目(GZKP-2021-39)。

语种：中文

中文关键词：镰形棘豆提取物;糖尿病肾脏病;足细胞;氧化应激;PI3K/AKT/Nrf2

外文关键词：oxytropis falcata extract;diabetic kidney disease;podocyte;oxidative stress;PI3K/AKT/Nrf2

摘要：目的:通过PI3K/AKT/Nrf2通路探讨镰形棘豆提取物含药血清对高糖诱导的足细胞氧化应激损伤的作用。方法:体外培养大鼠肾足细胞,筛选D-葡萄糖造模浓度和时间以及镰形棘豆提取物含药血清最佳给药浓度和时间;将细胞分为正常组、高糖组、高糖+空白血清组、镰形棘豆组、抑制剂组、镰形棘豆+抑制剂组,采用罗丹明染色及电镜观察足细胞形态、病理学改变,流式细胞术检测细胞凋亡率,划痕实验检测细胞迁移能力,免疫荧光及荧光探针法分别检测细胞p-AKT荧光表达和ROS水平,ELISA检测细胞上清液中MDA、NO、SOD、T-AOC含量,Western blot检测细胞p-PI3K/PI3K、p-AKT/AKT、Nrf2蛋白表达,RT-qPCR检测细胞Nrf2、HO-1、GCLM、SODmRNA表达。结果:筛选出45mmol/LD-葡萄糖诱导48h为最佳造模条件,含药血清1倍稀释及48 h为最佳给药浓度和时间。与正常组比较,高糖组细胞足突广泛融合、相互粘连,凋亡率、迁移能力和ROS水平均升高(P<0.01),p-AKT荧光表达降低(P<0.01),MDA、NO含量增加,SOD、T-AOC含量显著降低(P<0.01),p-PI3K/PI3K、p-AKT/AKT、Nrf2蛋白表达及Nrf2、HO-1、GCLM、SOD mRNA表达均降低(P<0.01)。与高糖组比较,各给药组细胞足突融合、相互粘连情况有不同程度的改善,凋亡率、迁移能力和ROS水平均降低(P<0.05,P<0.01),p-AKT荧光表达增加(P<0.01),MDA、NO含量下降,SOD、T-AOC含量升高(P<0.01),p-PI3K/PI3K、p-AKT/AKT、Nrf2蛋白及Nrf2、HO-1、GCLM、SOD mRNA表达均升高(P<0.05,P<0.01),而抑制剂组细胞ROS水平升高(P<0.01),p-AKT荧光表达降低(P<0.05),NO含量升高,T-AOC含量降低(P<0.01),p-PI3K/PI3K、p-AKT/AKT、Nrf2蛋白及HO-1、GCLM、SODmRNA表达降低(P<0.05,P<0.01)。与镰形棘豆组比较,镰形棘豆组+抑制剂组细胞凋亡率、迁移能力和ROS水平均升高(P<0.05,P<0.01),p-AKT荧光表达降低(P<0.01),MDA、NO含量增加,SOD、T-AOC含量降低(P<0.05,P<0.01),p-PI3K/PI3K、p-AKT/AKT、Nrf2蛋白及HO-1、GCLM、SODmRNA表达均降低(P<0.05,P<0.01)。结论:镰形棘豆提取物可能通过激活PI3K/AKT/Nrf2信号通路,从而改善高糖诱导的足细胞形态、结构和功能的改变,抑制氧化应激反应,达到保护足细胞的作用。
AIM:To investigate the effect of drugcontaining serum of oxytropis falcata extract on oxidative 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 modelling and the optimal concentration and time of administration in the drug-containing serum of oxytropis 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 microscopy were used to observe the morphological and pathological changes of podocyte,flow cytometry was used to detect apoptosis rate,and cell migration ability was detected by scratch test.Immunofluorescence and fluorescence probe were used to detect the fluorescence expression of p-AKT and ROS level of cells,ELISA was used to detect the content of MDA,NO,SOD and T-AOC in the supernatant 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 condition,and the 1-fold dilution of the medicated serum of oxytropis falcata extract for 48 hours was the best concentration and intervention time.Compared with the normal group,the foot processes of the high glucose group were widely fused and adhered 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 contents of MDA and NO were dramatically enhanced,and the contents of SOD and T-AOC were significantly downregulated(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 declined(P<0.01).Compared with high glucose group,foot process fusion and adhesion of podocytes in oxytropis falcata group and oxytropis falcata+inhibitor group were improved in varying degrees,apoptosis rate,migration ability and intracellular 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 improved(P<0.01),the fluorescence expression of p-AKT decreased(P<0.05),the content of NO upregulated,the content of T-AOC downregulated,and the expression of p-PI3K/PI3K,p-AKT/AKT,Nrf2 protein and HO-1,GCLM,SOD mRNA decreased in inhibitor group(P<0.05,P<0.01).Compared with oxytropis falcata group,the apoptosis rate,migration ability and intracellular ROS level of podocytes in oxytropis falcata+inhibitor group markedly increased(P<0.05,P<0.01),p-AKT fluorescence expression 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).CONCLUSION:Oxytropis falcata extract may protect podocytes by activating the PI3K/AKT/Nrf2 signaling pathway,thereby improving the morphological,structural and functional changes of podocytes induced by high glucose and inhibiting oxidative stress response.