LncRNA H19/miR-675-5p通過靶向Mfn2調(diào)控血管平滑肌細(xì)胞增殖和凋亡
發(fā)布時間:2021-03-25 11:35
目的:血管平滑肌細(xì)胞(vascular smooth muscle cell,VSMC)是構(gòu)成血管壁組織結(jié)構(gòu)及維持血管張力的主要細(xì)胞成分,VSMC異常增殖和凋亡是動脈粥樣硬化等血管重塑性疾病的病理學(xué)基礎(chǔ)。長鏈非編碼RNA(long non-coding RNA,lncRNA)是一類長度超過200個核苷酸、沒有蛋白質(zhì)編碼功能的RNA,廣泛參與了VSMC的功能調(diào)控。我們通過轉(zhuǎn)錄組測序分析發(fā)現(xiàn),大量lncRNA在大鼠內(nèi)膜增生血管中表達(dá)出現(xiàn)異常,其中l(wèi)ncRNA H19表達(dá)水平異常升高,H19是否參與了血管內(nèi)膜增生的調(diào)控?其分子機(jī)制如何?目前還不清楚。本研究擬從分子、細(xì)胞和整體水平,探究H19在VSMC增殖、凋亡和血管內(nèi)膜增生中的功能及分子機(jī)制。方法:1對大鼠行頸總動脈球囊內(nèi)皮剝脫術(shù),構(gòu)建血管內(nèi)皮損傷模型,28天后將內(nèi)膜增生血管與正常血管取材,進(jìn)行轉(zhuǎn)錄組測序分析。對差異mRNA進(jìn)行GO和Pathway分析,并通過qRT-PCR驗證。2體外培養(yǎng)VSMC并分別轉(zhuǎn)染siRNA Control、siRNA H19、mi R-675-5p inhibitor、siRNA H19+mi R-675-5p ...
【文章來源】:河北醫(yī)科大學(xué)河北省
【文章頁數(shù)】:74 頁
【學(xué)位級別】:碩士
【部分圖文】:
大鼠內(nèi)膜增生血管轉(zhuǎn)錄組測序分析
30圖 2 H19 促進(jìn) VSMC 增殖并抑制 VSMC 凋亡Fig.2 H19 promote the proliferation and inhibit the apoptosis of VSMC.
37圖 4 Mfn2 是 miR-675-5p 的調(diào)控靶標(biāo)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.
【參考文獻(xiàn)】:
期刊論文
[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)
本文編號:3099610
【文章來源】:河北醫(yī)科大學(xué)河北省
【文章頁數(shù)】:74 頁
【學(xué)位級別】:碩士
【部分圖文】:
大鼠內(nèi)膜增生血管轉(zhuǎn)錄組測序分析
30圖 2 H19 促進(jìn) VSMC 增殖并抑制 VSMC 凋亡Fig.2 H19 promote the proliferation and inhibit the apoptosis of VSMC.
37圖 4 Mfn2 是 miR-675-5p 的調(diào)控靶標(biāo)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.
【參考文獻(xiàn)】:
期刊論文
[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)
本文編號:3099610
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