摘要
目的探讨转染miR-885-5p 48 h后对人心肌成纤维细胞(HEH2)增殖的调控作用以及对基质金属蛋白酶2(MMP2)基因和特异性蛋白1(SP1)基因的表达的影响,为临床心血管疾病研究提供研究线索。方法培养HEH2细胞待细胞长到60%时通过脂质体转染方法对细胞转染miR-885-5p和阴性对照(NC),48h后用CCK8细胞活力试剂盒来检查细胞增殖情况。实时荧光定量PCR法检查miR-885-5p对MMP2和SP1的RNA表达的影响。结果CCK8结果表明转染48 h后阴性对照组细胞活率为(100±8)%,miR-885-5p组细胞活率为(139±16)%,提示miR-885-5p促进HEH2细胞的增殖(n=6,P<0.05),HEH2细胞转染miR-885-5p后,心肌重构相关基因MMP2和SP1的miRNA表达增高(P<0.05)。结论miR-885-5p能够促进人心肌成纤维细胞的增殖,同时促进细胞中MMP2和SP1基因的表达,提示miR-885-5p抑制剂可能是心肌纤维化相关疾病的新策略。
Objective To investigate the regulation of human cardiac fibroblasts(HEH2)by transfection of miR-885-5 p and the effects on the expression of the protease 2(MMP2)gene and the SP1 gene to provide clues for clinical cardiovascular disease research.Methods HEH2 cells were transfected into miR-885-5 p and negative control(NC)by liposome transfection when cells were grown to 60%.After 48 hours,CCK8 cell viability kit was used to check cell proliferation.The effect of miR-885-5 p on RNA expression of MMP2 and SP1 was examined by real-time fluorescent quantitative PCR.Results The results of CCK8 showed that the cell viability of the negative control group was(100±8)% after 48 hours of transfection and the cell viability of miR-885-5 p group was(139±16)%,suggesting that miR-885-5 p promotes the proliferation of HEH2 cells(n=6.P<0.05);the expression of miRNA in myocardial remodeling related genes MMP2 and SP1 was increased after HEH2 cells were transfected with miR-885-5 p(P<0.05).Conclusion MiR-885-5 p can promote the proliferation of human cardiac fibroblasts and promote the expression of MMP2 and SP1 genes in cells,suggesting that miR-885-5 p inhibitor may be a new strategy for myocardial fibrosis-related diseases.
作者
张新生
刘超群
刘梦瑶
滕松
许何丽
窦文静
王喜燕
王玉华
张越
ZHANG Xin-sheng;LIU Chao-qun;LIU Meng-yao;TENG Song;XU He-li;DOU IVen-jing;IVANG Xi-yan;WANG Yu-hua;ZHANG Yue(Department of Occupational Health,School of Public Health,Shenyang Medical College,Shenyang 110001,China)
出处
《中国心血管病研究》
CAS
2019年第12期1084-1087,共4页
Chinese Journal of Cardiovascular Research
基金
国家自然科学基金(81070128)
教育部留学启动基金(20121707)
辽宁省重点研发计划(2018225013)
辽宁省杰出青年学者成长计划(教育厅优秀人才项目No.L2011112)
沈阳市“高层次创新人才计划”项目(RC170408)
沈阳中青年高层次人才创新支持计划(RC180379)
沈阳医学院优秀人才项目(20113067)
沈阳医学院硕士研究生科研创新基金项目(Y20190501、Y20190507)~~
