miRNA-7658-5p通过调控氧化应激反应促进同型半胱氨酸诱导的巨噬细胞凋亡OA

AIM:To investigate the function and regulatory mechanism of microRNA-7658-5p(miRNA-7658-5p)in homocysteine(Hcy)-induced apoptosis of mouse macrophages(RAW264.7 cells),as well as its effects on oxida-tive stress and mitochondrial membrane potential.METHODS:Mouse RAW264.7 macrophages were cultured in vitro,and divided into a control group(Hcy:0 µmol/L)and an Hcy-treated group(Hcy:100 µmol/L).Flow cytometry was used to detect the intracellular level of reactive oxygen species(ROS).JC-1 fluorescent staining was employed to observe changes in mitochondrial structure and membrane potential.Western blot was performed to detect the expression levels of apoptosis-related proteins B-cell lymphoma 2(Bcl-2).Bcl-2-associated X protein(Bax);and qRT-PCR was used to de-termine the transcriptional level of miRNA-7658-5p.Furthermore,the cells were transfected with miRNA-7658-5p mim-ic,inhibitor,and negative control,and the above indicators were re-analyzed.Finally,Western blot and flow cytometry were used to detect the expression of apoptosis-related proteins and the cell apoptosis rate.RESULTS:Compared with the control group,the Hcy-treated group showed significantly increased expression of miRNA-7658-5p,expression of Bax protein,and intracellular ROS accumulation(P<0.01),while the expression of Bcl-2 protein and mitochondrial mem-brane potential were significantly decreased(P<0.01).Functional verification experiments showed that overexpression of miRNA-7658-5p enhanced Hcy-induced macrophage apoptosis(P<0.01),whereas inhibition of its expression significant-ly alleviated apoptosis(P<0.01).CONCLUSION:miRNA-7658-5p promotes Hcy-induced macrophage apoptosis by regulating oxidative stress.