中国针灸2026，Vol.46Issue(4)：580-588,9.DOI:10.13703/j.0255-2930.20250624-k0004
基于Keap1/Nrf2/GPX4通路介导铁死亡探讨电针调控功能性消化不良大鼠十二指肠损伤的机制
Mechanism of electroacupuncture for attenuating duodenal injury in rats with functional dyspepsia via Keap1/Nrf2/GPX4 pathway-mediated ferroptosis
摘要
Abstract
Objective To investigate the mechanism of electroacupuncture(EA)at"Zusanli"(ST36)and"Taichong"(LR3)for attenuating duodenal injury in rats with functional dyspepsia(FD)based on ferroptosis mediated by Kelch-like epichlorohydrin-associated protein 1(Keap1)/nuclear factor E2-related factor 2(Nrf2)/glutathione peroxidase 4(GPX4)pathway.Methods A total of 21 SPF-grade SD male rats were randomly divided into a normal group,a model group and an EA group,7 rats in each one.In the model and EA groups,FD(liver stagnation and spleen deficiency)models were prepared by dual-factor combination method,using mild tail clamping stimulation and intermittent diet.After model preparation,in the EA group,EA was operated at bilateral"Zusanli"(ST36)and"Taichong"(LR3),with a disperse-dense wave,a frequency of 2 Hz/100 Hz and for 20 min each session,once a day,and the duration of intervention was composed of 14 days.The general condition of the rats was observed.The gastric emptying rate and intestinal propulsion rate were measured.The pathomorphologic changes of duodenal tissue were observed by HE staining method;the positive expression of Keap1,Nrf2 and GPX4 in duodenal tissue was detected using immunohistochemistry and the positive expression of reactive oxygen species(ROS)in duodenal tissue was detected using immunofluorescence.The levels of ferrous ions(Fe2+)in duodenal tissue,and malondialdehyde(MDA),reduced glutathione(GSH)and superoxide dismutase(SOD)in serum were detected using biochemical methods,ATP level in serum was detected using ELISA,and the mRNA expression of Keap1,Nrf2,and GPX4 in duodenal tissue was detected by real-time fluorescence quantitative PCR(RT-qPCR);and the protein expression of Keap1,Nrf2,GPX4,acyl-CoA synthase long-chain family 4(ACSL4)and solute carrier family 7 member 11(SLC7A11)was detected using Western blot.Results Compared with the normal group,the rats in the model group exhibited the decrease in food intake,body weight,gastric emptying rate and intestinal propulsion rate(P＜0.05).In the duodenal tissue,the intestinal villi were disordered,and the inflammatory cells infiltrated.The positive expression of ROS,and the contents of Fe2+and MDA were elevated(P＜0.05),the levels of GSH,SOD and ATP decreased(P＜0.05),the mRNA expression of Nrf2 and GPX4 and the protein expression of Nrf2,GPX4 and SLC7A11 were reduced(P＜0.05),while the mRNA expression of Keap1 and the protein expression of Keap1 and ACSL4 were elevated(P＜0.05).When compared with the model group,the rats in the EA group showed the increase in food intake,body weight,gastric emptying rate and intestinal propulsion rate(P＜0.05).In the duodenal tissue,the structure of the intestinal mucosa was intact basically,without inflammatory cell infiltration in the lamina propria.The positive expression of ROS,and the contents of Fe2+and MDA were dropped(P＜0.05),the contents of GSH,SOD and ATP rose(P＜0.05),the mRNA expression of Nrf2 and GPX4 and the protein expression of Nrf2,GPX4 and SLC7A11 were elevated(P＜0.05),while the mRNA expression of Keap1 and the protein expression of Keap1 and ACSL4 were reduced(P＜0.05).Conclusion Electroacupuncture at"Zusanli"(ST36)and"Taichong"(LR3)alleviates lipid peroxidation and oxidative stress injury in the duodenum of FD rats,maintains intracellular iron homeostasis,and inhibits ferroptosis probably by regulating Keap1/Nrf2/GPX4 pathway,so as to attenuate FD.关键词
功能性消化不良/电针/铁死亡/氧化应激/十二指肠损伤/Keap1/Nrf2/GPX4Key words
functional dyspepsia/electroacupuncture/ferroptosis/oxidative stress/duodenal injury/Keap1/Nrf2/GPX4引用本文复制引用
冯依凡,梁婷婷,田跃龙,韩磊,刘继东,曲怡,薛亚楠,王建波,李阳,王天朗,王熙涵,张家豪..基于Keap1/Nrf2/GPX4通路介导铁死亡探讨电针调控功能性消化不良大鼠十二指肠损伤的机制[J].中国针灸,2026,46(4):580-588,9.基金项目
国家自然科学基金面上项目:82474634 （）
辽宁省科技计划联合计划项目:2024JH2/102600199 （）
