LncRNA H19/miR-675-5p通过靶向Mfn2调控血管平滑肌细胞增殖和凋亡
发布时间:2021-03-25 11:35
目的:血管平滑肌细胞(vascular smooth muscle cell,VSMC)是构成血管壁组织结构及维持血管张力的主要细胞成分,VSMC异常增殖和凋亡是动脉粥样硬化等血管重塑性疾病的病理学基础。长链非编码RNA(long non-coding RNA,lncRNA)是一类长度超过200个核苷酸、没有蛋白质编码功能的RNA,广泛参与了VSMC的功能调控。我们通过转录组测序分析发现,大量lncRNA在大鼠内膜增生血管中表达出现异常,其中lncRNA H19表达水平异常升高,H19是否参与了血管内膜增生的调控?其分子机制如何?目前还不清楚。本研究拟从分子、细胞和整体水平,探究H19在VSMC增殖、凋亡和血管内膜增生中的功能及分子机制。方法:1对大鼠行颈总动脉球囊内皮剥脱术,构建血管内皮损伤模型,28天后将内膜增生血管与正常血管取材,进行转录组测序分析。对差异mRNA进行GO和Pathway分析,并通过qRT-PCR验证。2体外培养VSMC并分别转染siRNA Control、siRNA H19、mi R-675-5p inhibitor、siRNA H19+mi R-675-5p ...
【文章来源】:河北医科大学河北省
【文章页数】:74 页
【学位级别】:硕士
【部分图文】:
大鼠内膜增生血管转录组测序分析
30图 2 H19 促进 VSMC 增殖并抑制 VSMC 凋亡Fig.2 H19 promote the proliferation and inhibit the apoptosis of VSMC.
37图 4 Mfn2 是 miR-675-5p 的调控靶标Fig.4 Mfn2 is the regulatory target of miR-675-5pA. The miR-675-5p binding site in the 3'UTR of Mfn2. B. The luciferase activity ofpGL3-MFN2-3'UTR reporter gene. *P < 0.05, compared with mutant control. C. Westernblot analysis shows the expression of Mfn2 in VSMC at 48 hours after transfection,normalized with -actin. D. Western blot analysis of Mfn2 protein expression level inVSMC at 48 hours after transfection. *P < 0.05, compared with control.
【参考文献】:
期刊论文
[1]A bioinformatics method for predicting long noncoding RNAs associated with vascular disease[J]. LI JianWei,GAO Cheng,WANG YuChen,MA Wei,TU Jian,WANG JunPei,CHEN ZhenZhen,KONG Wei,CUI QingHua. Science China(Life Sciences). 2014(08)
[2]miRNAs and lncRNAs in vascular injury and remodeling[J]. SONG XiaoWei,SHAN DongKai,CHEN Jian,JING Qing. Science China(Life Sciences). 2014(08)
本文编号:3099610
【文章来源】:河北医科大学河北省
【文章页数】:74 页
【学位级别】:硕士
【部分图文】:
大鼠内膜增生血管转录组测序分析
30图 2 H19 促进 VSMC 增殖并抑制 VSMC 凋亡Fig.2 H19 promote the proliferation and inhibit the apoptosis of VSMC.
37图 4 Mfn2 是 miR-675-5p 的调控靶标Fig.4 Mfn2 is the regulatory target of miR-675-5pA. The miR-675-5p binding site in the 3'UTR of Mfn2. B. The luciferase activity ofpGL3-MFN2-3'UTR reporter gene. *P < 0.05, compared with mutant control. C. Westernblot analysis shows the expression of Mfn2 in VSMC at 48 hours after transfection,normalized with -actin. D. Western blot analysis of Mfn2 protein expression level inVSMC at 48 hours after transfection. *P < 0.05, compared with control.
【参考文献】:
期刊论文
[1]A bioinformatics method for predicting long noncoding RNAs associated with vascular disease[J]. LI JianWei,GAO Cheng,WANG YuChen,MA Wei,TU Jian,WANG JunPei,CHEN ZhenZhen,KONG Wei,CUI QingHua. Science China(Life Sciences). 2014(08)
[2]miRNAs and lncRNAs in vascular injury and remodeling[J]. SONG XiaoWei,SHAN DongKai,CHEN Jian,JING Qing. Science China(Life Sciences). 2014(08)
本文编号:3099610
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