安徽医科大学学报2025，Vol.60Issue(12)：2207-2214,8.DOI:10.19405/j.cnki.issn1000-1492.2025.12.003
LncRNA RMRP通过调控miR-766-5p对氧糖剥夺/再灌注诱导的小鼠HL-1心肌细胞铁死亡的影响
Effect of LncRNA RMRP on ferroptosis induced by oxygen glucose deprivation/reperfusion in mouse HL-1 cardiomyocytes by regulating miR-766-5p
摘要
Abstract
Objective To investigate the effect and mechanism of long non-coding RNA RMRP(LncRNA RMRP)on oxygen-glucose deprivation/reperfusion(OGD/R)-induced ferroptosis in mouse HL-1 cardiomyocytes by regula-ting miR-766-5p.Methods HL-1 cells were cultured in vitro,and OGD/R models were established.The expres-sion levels of LncRNA RMRP in HL-1 cells at various reperfusion time points were subsequently quantified using qRT-PCR.The LncRNA RMRP small RNA interference fragment(si-RMRP)and its corresponding negative con-trol(si-NC),as well as the miR-766-5p inhibitor and its respective negative control(inhibitor-NC),were trans-fected into HL-1 cells.Subsequently,the cells were subjected to OGD/R treatment.CCK-8 assay was employed to evaluate cell viability.Assay kits were employed to measure the levels of lactate dehydrogenase(LDH)in the cell supernatant,as well as the intracellular levels of malondialdehyde(MDA),superoxide dismutase(SOD),gluta-thione(GSH),and ferrous ion(Fe2+).qRT-PCR analysis was conducted to assess the expression levels of Ln-cRNA RMRP and miR-766-5p.Western blot analysis was conducted to assess the expression levels of proteins asso-ciated with ferroptosis including GPX4,SLC7A11,and FTH1.Dual-luciferase reporter assays were performed to investigate the sponge adsorption relationship between LncRNA RMRP and miR-766-5p.Results As reperfusion time extended,the expression level of LncRNA RMRP in cells progressively increased(P＜0.01).Treatment with OGD/R significantly inhibited the viability of HL-1 cells,reduced the expression of miR-766-5p(P＜0.01),ele-vated the levels of LDH in the supernatant,as well as MDA and Fe2+levels within the cells,and decreased the ac-tivities of SOD and GSH in cells(P＜0.01).Additionally,OGD/R treatment downregulated the protein expres-sion levels of GPX4,SLC7A11,and FTH1(P＜0.01).Silencing LncRNA RMRP reversed these effects by enhan-cing the viability of HL-1 cells,increasing miR-766-5p expression(P＜0.01),reducing LDH in the supernatant,as well as MDA and Fe2+levels within the cells,and promoting SOD and GSH activities in cells(P＜0.01).Fur-thermore,silencing LncRNA RMRP upregulated the protein expression levels of GPX4,SLC7A11,and FTH1(P＜0.01).The dual-luciferase reporter assay confirmed that LncRNA RMRP could regulate the expression of miR-766-5p through a sponge adsorption mechanism.Partial inhibition of miR-766-5p inhibitor expression could mitigate the improvement effect caused by LncRNA RMRP silencing on OGD/R-induced ferroptosis in HL-1 cells.Conclusion Silencing LncRNA RMRP inhibits OGD/R-induced ferroptosis in HL-1 cells,potentially through the sponge-me-diated regulation of miR-766-5p expression.关键词
氧糖剥夺/再灌注/心肌/HL-1细胞/铁死亡/长链非编码RNA RMRP/miR-766-5pKey words
oxygen-glucose deprivation/reperfusion/myocardium/HL-1 cells/ferroptosis/long non-coding RNA RMRP/miR-766-5p分类
医药卫生引用本文复制引用
何蕾,孙兴兰,吴应兴,许源,彭翔,胡晨恺..LncRNA RMRP通过调控miR-766-5p对氧糖剥夺/再灌注诱导的小鼠HL-1心肌细胞铁死亡的影响[J].安徽医科大学学报,2025,60(12):2207-2214,8.基金项目
江西省自然科学基金青年基金项目(编号:20232BAB216008) （编号:20232BAB216008）
中国国家留学基金项目(编号:202506820050) Natural Science Foundation of Jiangxi Province(No.20232BAB216008) （编号:202506820050）
Funding from China Scholarship Council(No.202506820050) （No.202506820050）
