湖南中医药大学学报2025，Vol.45Issue(3)：425-431,7.DOI:10.3969/j.issn.1674-070X.2025.03.005
补肾活血汤含药血清调控髓核细胞外泌体中miR-222-3p抑制髓核细胞凋亡的研究
Effects of BuShen Huoxue Decoction-medicated serum on regulating miR-222-3p expression in exosomes and inhabiting nucleus pulposus cell apoptosis
摘要
Abstract
Objective To investigate the inhibitory effects of Bushen Huoxue Decoction-medicated serum on nucleus pulposus cell apoptosis by analyzing its regulation of miR-222-3p expression in exosomes derived from nucleus pulposus cell.Methods(1)Nucleus pulposus cells were randomly divided into PBS group,blank serum group,and medicated serum group.The blank serum group was supplemented with 10%normal serum,the medicated serum gro up with 10%Bushen Huoxue Decoction-medicated serum,and the PBS group with an equal volume of PBS.After 48 h,samples were collected,and exosomes were extracted.Transmission electron microscopy was used to identify the morphological structure of exosomes.Western blot was used to determine protein expressions of lysosome-associated membrane protein 3(CD63)and tumor susceptibility gene 101(TSG101)in exosomes.Real-time PCR was applied to examine the miR-222-3p expression in exosomes.(2)Exosomes were co-cultured with tumor necrosis factor-α(TNF-α)to induce nucleus pulposus cell models.The nucleus pulposus cells were divided into blank group,model group,blank exosome group,medicated exosome group,and Anti-miR-222-3p exosome group.After 24 h of co-culture,the apoptosis rate of nucleus pulposus cells was measured by flow cytometry,the expression of miR-222-3p in nucleus pulposus cells was determined by real-time PCR,and the protein expressions of p53,cytochrome C(Cytc),and cysteine-aspartic acid protegase 1(Caspase-1)were checked by Western blot among different groups.Results(1)Compared with the blank serum group,the medicated serum group showed increased number of exosomes(P<0.05),elevated relative protein expression levels of exosomal marker proteins CD63 and TSG101(P<0.05),and decreased miR-222-3p expression in exosomes derived from nucleus pulposus cells.(2)Compared with the model group and the blank exosome group,the apoptosis rate of nucleus pulposus cells further decreased,the miR-222-3p expression in nucleus pulposus cells decreased(P<0.05),and the relative protein expression levels of p53,Cytc,and Caspase-1 decreased(P<0.05)in the medicated exosome group and the Anti-miR-222-3p exosome group.Conclusion(1)Bushen Huoxue Decoction-medicated serum can effectively prompt the secretion of exosomes by rat nucleus pulposus cells and inhibit miR-222-3p expression in exosomes derived from these cells.(2)Bushen Huoxue Decoction-medicated exosomes can alleviate the pathological damage of degenerative nucleus pulposus cells induced by TNF-α and reduce nucleus pulposus cell apoptosis.The mechanism may be closely associated with the inhibition of expressions of miR-222-3p as well as apoptosis-related proteins p53,Cytc,and Caspase-1.关键词
补肾活血汤/髓核细胞/外泌体/miR-222-3p/细胞凋亡Key words
Bushen Huoxue Decoction/nucleus pulposus cells/exosomes/miR-222-3p/apoptosis分类
医药卫生引用本文复制引用
谭婉俊,吴官保,彭兴宁,冯帅华..补肾活血汤含药血清调控髓核细胞外泌体中miR-222-3p抑制髓核细胞凋亡的研究[J].湖南中医药大学学报,2025,45(3):425-431,7.基金项目
湖南省自然科学基金面上项目(2022JJ30025) （2022JJ30025）
湖南省卫生健康委员会一般课题(W20243066). （W20243066）
