宁夏医科大学学报2024,Vol.46Issue(1) :7-14.DOI:10.16050/j.cnki.issn1674-6309.2024.01.002

Hcy促血管平滑肌细胞增殖迁移相关的miRNAs筛选研究

Screening of miRNAs Associated with Hcy Promoting Proliferation and Migration of Vascular Smooth Muscle Cells

马星 桂娜 马婷 王秀玉 莫廷润 张鸣号
宁夏医科大学学报2024,Vol.46Issue(1) :7-14.DOI:10.16050/j.cnki.issn1674-6309.2024.01.002

Hcy促血管平滑肌细胞增殖迁移相关的miRNAs筛选研究

Screening of miRNAs Associated with Hcy Promoting Proliferation and Migration of Vascular Smooth Muscle Cells

马星 1桂娜 2马婷 3王秀玉 3莫廷润 3张鸣号1
扫码查看

作者信息

  • 1. 宁夏医科大学基础医学院,银川 750004;国家卫生健康委员会代谢性心血管疾病研究重点实验室,银川 750004;宁夏血管损伤与修复研究重点实验室,银川 750004
  • 2. 宁夏医科大学基础医学院,银川 750004;宁夏贺兰县人民医院,贺兰 750200
  • 3. 宁夏医科大学基础医学院,银川 750004
  • 折叠

摘要

目的 明确同型半胱氨酸(homocysteine,Hcy)对人源血管平滑肌细胞(vascular smooth muscle cell,VSMC)增殖迁移的影响以及分析Hcy干预下VSMC增殖迁移相关差异miRNAs,为探讨Hcy促VSMC增殖迁移的分子机制提供依据.方法 体外培养VSMC,分为Control组和 100 μmol·L-1 Hcy组.采用CCK-8 法测量VSMC的增殖活性;采用划痕实验分析VSMC在 0h和 48h的迁移情况.高通量测序分析各组VSMC中差异化表达的miRNAs,采用miRanda和TargetScan软件进行靶基因预测,使用BLAST软件将预测靶基因序列与GO和KEGG数据库比对,分析靶基因的功能,获得与VSMC增殖和迁移相关的靶基因信息.通过RT-qPCR对其中表达上调的miRNAs进行验证.结果 Hcy干预能增加VSMC的增殖活力(P<0.01),且VSMC的划痕面积减少(P<0.01).两组细胞共检测到 576 个miRNAs,其中已知miRNAs 404 个,新预测miRNAs 172 个.与Control组相比,Hcy组存在 88 个差异表达的miRNAs,其中表达上调的 20 个,表达下调的 68 个.与VSMC增殖和迁移相关的miRNAs有 20 个,表达上调的包括miRNA39、miR NA206 和miRNA212-5p,表达下调的包括miR NA35等17个.RT-qPCR结果显示,Hcy组miRNA212-5p的表达上调(P<0.05).结论 Hcy致VSMC增殖迁移过程中存在差异表达的miRNAs,miR NA212-5p可能参与了Hcy对VSMC的作用.

Abstract

Objective To determine the effects of homocysteine(Hcy)on proliferation and migration of human vascular smooth muscle cell(VSMC)and analyze the differences related to the proliferation and migration of VSMC under the intervention of Hcy.Methods VSMC was cultured in v itro and divided into Control group and 100 μmol·L-1 Hcy intervention group.The proliferative activity of VSMC was measured by CCK-8 method.The migration of VSMC at 0 h and 48 h was analyzed by scratch test.The differentiated expression of miRNAs in VSMC in each group was analyzed by high-throughput sequencing,target gene prediction was performed us-ing miRanda and TargetScan software,and the function of target genes was analyzed by BLAST software com-pared with GO and KEGG databases to obtain the target gene information related to VSMC proliferation and mi-gration.The up-regulated miRNAs were verified by RT-q PCR.Results Hcy intervention could significantly increase the proliferative activity of VSMC(P<0.01),and the scratch area of VSMC was significantly reduced(P<0.01),suggesting that 100 μmol·L-1 Hcy intervention could promote the proliferation and migration of VSMC,which was consistent with the results of previous studies.In terms of mechanism,a total of 576 miRNAs were detected in both groups of cells,of which 404 were known and 172 were newly predicted.Compared with the Control group,there were 88 differentially expressed miRNAs in the Hcy group,of which 20 were up-regu-lated and 68 were down-regulated.Among them,there were 20 miRNAs related to VSMC proliferation and mi-gration,including miR NA39,miR NA206,and miRNA212-5p whose expression was up-regulated,and 17 miR NA35 whose expression was down-regulated.The results of RT-qPCR showed that the expressions of miR-NA212-5p was significantly increased in the Hcy group(P<0.05).Conclusion There are differentially ex-pressed miRNAs in the proliferation and migration of VSMC induced by Hcy.miR NA212-5p may be involved in the effect of Hcy on VSMC.

关键词

微小RNA/靶基因预测/高通量测序/同型光胱氨酸/血管平滑肌细胞

Key words

miRNAs/target gene prediction/high-throughput sequencing/homocysteine/vascular smooth mus-cle cell

引用本文复制引用

基金项目

宁夏回族自治区重点研发计划项目(2021BEG03093)

宁夏回族自治区科技惠民专项项目(2022CMG03036)

宁夏自然科学基金项目(2022AAC03130)

宁夏医科大学校级科研项目(XT2022028)

出版年

2024
宁夏医科大学学报
宁夏医科大学

宁夏医科大学学报

CSTPCD
影响因子:0.84
ISSN:1674-6309
参考文献量19
段落导航相关论文