灵芝miRNA对人皮肤成纤维细胞衰老的作用研究  

Effect of Ganoderma lucidum miRNA on the senescence of human skin fibroblasts

作　　者：余雯斌 徐晓渊 盛清[1] YU Wenbin;XU Xiaoyuan;SHENG Qing(College of Life Sciences and Medicine,Zhejiang Sci-Tech University,Hangzhou 310018,China)

机构地区：[1]浙江理工大学生命科学与医药学院,杭州310018

出　　处：《浙江理工大学学报（自然科学版）》2024年第1期120-129,共10页Journal of Zhejiang Sci-Tech University(Natural Sciences)

基　　金：浙江省基础公益研究计划项目(LGF18H250004)。

摘　　要：为探究灵芝中miRNA对细胞衰老的干预作用及影响,利用过氧化氢(H_(2)O_(2))诱导人皮肤成纤维细胞(Human skin fibroblasts,HSF)构建了细胞衰老模型,将灵芝特有的miRNA Glu-miR-01和Glu-miR-03转染HSF衰老细胞,分析细胞衰老相关氧化应激指标,并通过Western blotting检测衰老相关蛋白表达水平的变化。结果显示:构建HSF细胞衰老模型的最适条件为H_(2)O_(2)浓度0.8mmol/L,诱导时间3h;Glu-miR-01和Glu-miR-03均能促使衰老细胞中超氧化物歧化酶(Superoxide dismutase,SOD)和总抗氧化能力(Total antioxidant capacity,T-AOC)水平上升,丙二醛(Malondialdehyde,MDA)含量下降,细胞衰老相关β-半乳糖苷酶(Senescence associated-β-Galactosidase,SA-β-gal)染色阳性率降低,线粒体膜电位升高;衰老信号通路中P53、P21和P16蛋白的表达显著降低,Rb蛋白表达显著升高。该结果表明灵芝中Glu-miR-01和Glu-miR-03对HSF衰老细胞具有保护作用,并对衰老相关的p53/p21/Rb信号通路具有调控作用,为阐明灵芝miRNA对细胞衰老的分子作用机制提供了理论依据。To investigate the intervention effect and impact of miRNA in Ganoderma lucidum(G.lucidum)on cell aging,we used hydrogen peroxide(H_(2)O_(2))to induce human skin fibroblasts(HSF)and constructed a cell senescence model.The HSF senescent cells were transfected with miRNA Glu-miR-01 and Glu-miR-03,which were unique to G.lucidum,and the oxidative stress indexes related to cell aging were analyzed.The change of the expression levels of age-related proteins was detected by Western blotting.The results indicated that the optimal conditions for establishing the HSF cell senescence model were 0.8mmol/L H_(2)O_(2)and 3hinduction time.Both Glu-miR-01and Glu-miR-03could enhance the activity of superoxide dismutase(SOD)and total antioxidant capacity(T-AOC),reduce the content of malondialdehyde(MDA),decrease the positive rate of senescence-β-galactosidase(SA-β-gal)staining and increase mitochondrial membrane potential in senescence cells.In the aging signaling pathway,the expression of P53,P21and P16proteins was significantly downregulated,while the expression of Rb protein was significantly upregulated.These findings suggest that Glu-miR-01and Glu-miR-03from G.lucidum exert protective effects on HSF senescent cells,and regulate the p53/p21/Rb signaling pathway associated with aging.This provides a theoretical basis for elucidating the molecular mechanism of G.lucidum miRNA on cell senescence.

关 键 词：灵芝 MIRNA 过氧化氢 氧化应激 细胞衰老 p53/p21/Rb信号通路 

分 类 号：Q936[生物学—微生物学]