Chuangling Ye Ameliorates Diabetic Foot Ulcers by Regulating Ferroptosis via the Nrf2/SLC7A11/GPX4 Signaling Pathway:Mechanisms and Therapeutic Effects
10.14148/j.issn.1672-0482.2025.1344
- VernacularTitle:疮灵液通过Nrf2/SLC7A11/GPX4信号通路调控铁死亡改善糖尿病足溃疡的作用及机制研究
- Author:
Ya ZHAO
1
;
Xiao FENG
;
Cunyu ZHANG
;
Yue CHEN
;
Chaoqun MA
Author Information
1. 南京中医药大学附属医院,江苏 南京 210029;南京中医药大学第一临床医学院,江苏 南京 210023
- Publication Type:Journal Article
- Keywords:
Chuangling Ye;
diabetic foot ulcer;
Nrf2/SLC7A11/GPX4 pathway;
ferroptosis
- From:
Journal of Nanjing University of Traditional Chinese Medicine
2025;41(10):1344-1355
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE To investigate the effects of Chuangling Ye(CLY)on diabetic foot ulcer(DFU)model mice based on the nuclear factor erythroid 2-related factor 2(Nrf2)/solute carrier family 7 member 11(SLC7A11)/glutathione peroxidase 4(GPX4)pathway and explore its potential mechanism.METHODS In animal experiments,streptozotocin(STZ)-induced diabetic mice with full-thickness skin defects were divided into control,model,positive control(recombinant human epidermal growth factor,rhEGF),and CLY low-and high-dose groups(n=10).After 14 days of intervention,wound healing rate was measured.Serum inflammatory factors(IL-6,TNF-α,IL-1β)were detected by ELISA,while oxidative stress markers(SOD,MDA,GSH)and tissue ferrous iron(Fe2+)levels were measured by colorimetric assays.Mitochondrial ultrastructure was observed via transmission electron microscopy(TEM).Nrf2,SLC7A11,and GPX4 expression in wound tissues were analyzed by qPCR and Western blot.In cell experiments,a high glucose(HG)-induced ferroptosis model in human umbilical vein endothelial cells(HUVECs)was established and divided into control,HG,CLY,and CLY combined with pathway inhibitors(ML385,Erastin,RSL3)groups.CCK-8 was used to detect cell via-bility,FerroOrange was used to detect Fe2+content,DCFH-DA and C11-BODIPY 581/591 probes were used to detect intracellular ROS and lipid ROS content,and Western blot was used to detect the expression of proteins such as Nrf2/SLC7A11/GPX4.RESULTS Animal experiments showed that compared with the model group,the wound healing rate in the low-and high-dose CLY groups was significantly improved(P<0.01);the serum IL-6,IL-1β,and TNF-α levels were significantly decreased(P<0.01);the MDA con-tent was significantly reduced(P<0.01),and the SOD activity and GSH content were significantly restored(P<0.05).Colorimetric a-nalysis showed that the low-and high-dose CLY significantly reduced the abnormally elevated Fe2+levels in DFU wounds(P<0.05,P<0.001).Electron microscopy showed that the mitochondrial cristae structure was improved in the low-and high-dose groups.qPCR showed that high-dose CLY upregulated the expression levels of Rno-Nfe2l2(Nrf2),Rno-Slc7a11,Rno-Gpx4,and Rno-Acsl4 mR-NA,and inhibited the expression level of Rno-Acsl4 mRNA(P<0.001).Western blot results showed that CLY upregulated the expres-sion of Nrf2,SLC7A11,GPX4,and FTH1 in DFU wound tissues(P<0.01),while downregulated the level of ACSL4(P<0.01).Cell experiments showed that treatment with CLY increased the survival rate of high glucose-stimulated HUVECs(P<0.001).However,the protective effect of CLY was significantly inhibited after the addition of ML385,Erastin,or RSL3(P<0.01).Treatment with CLY significantly decreased the Fe2+content(P<0.001),which was reversed by ML385,Erastin,or RSL3.The levels of ROS and lipid ROS were also significantly reduced(P<0.001),while the addition of ML385,Erastin,or RSL3 partially weakened the antioxidant effect of CLY.Western blot results showed that CLY significantly upregulated the expression of Nrf2、SLC7A11 and GPX4(P<0.001)and downregulated the expression of 4-HNE and COX2(P<0.01),and ML385,Erastin,or RSL3 could reverse this protective effect.CONCLUSION CLY promotes DFU healing by activating the Nrf2/SLC7A11/GPX4 pathway to inhibit ferroptosis,mitigate oxidative stress,and suppress inflammation,providing a novel therapeutic target for DFU.
