miR-126多基因靶向调控血管再生的研究
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摘要
目的:探讨microRNA miR-126多基因靶向调控血管再生的作用。方法:该实验将miR-126转染人脐静脉血管内皮细胞(HUVEC)后,随机分成A,B两组(A实验组:miRNA+HUVEC;B对照组:negative-miRNA control+HUVEC),荧光标记法和Western bolt检测2组细胞中VEGF和ANG-1 mRNA和蛋白表达, Matrigel实验检测2组细胞成管能力。裸鼠体内实验验证miR-126促进血管再生的能力。结果:A组细胞miRNA-126高表达,VEGF和ANG-1 mRNA和蛋白的表达增强(P<0.05),体外成管腔能力增强(P<0.05);在裸鼠体内异位再生血管环境中血管形成率增强(P<0.05)。结论:miR-126可促进血管化形成,其机制与增强相关靶基因表达升高有关。
Objective: To study the effects of miR-126 in the vasclurization by regulating multiple angiogenesis genes of human umbilical vein endothelial cells(HUVECs). Methods: miR-126 was transfected into HUVECs. The cells were divided into group A(experimental group: micro RNA-126+HUVECs) and B(control group: microRNA negative control+HUVECs).The expression of VEGF and ANG-1 mRNA and protein of the 2 groups was examined by fluorimetry and Western blot. Results: miR-126 over expression was observed in group A, the tube formation ability in vitro and stable micro-vessel formation in vivo were stronger in group A than in group B(P<0.05). The angiogenesis in nude mice of group A was more than that of group B. Conclusion: MiR-126 may induce angiogenesis and vascularization of HUVECs by regulating multiple angiogenesis genes.
Objective: To study the effects of miR-126 in the vasclurization by regulating multiple angiogenesis genes of human umbilical vein endothelial cells(HUVECs). Methods: miR-126 was transfected into HUVECs. The cells were divided into group A(experimental group: micro RNA-126+HUVECs) and B(control group: microRNA negative control+HUVECs).The expression of VEGF and ANG-1 mRNA and protein of the 2 groups was examined by fluorimetry and Western blot. Results: miR-126 over expression was observed in group A, the tube formation ability in vitro and stable micro-vessel formation in vivo were stronger in group A than in group B(P<0.05). The angiogenesis in nude mice of group A was more than that of group B. Conclusion: MiR-126 may induce angiogenesis and vascularization of HUVECs by regulating multiple angiogenesis genes.
引文
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[3] Carmeliet P.Mechanisms of angiogenesis and arteriogenesis[J].Nat Med,2000,6(4):389-395.
[4] Chen RR,Silva EA,Yuen WW,et al.Spatio- temporal VEGF and PDGF delivery patterns blood vessel formation and maturation[J].Pharm Res,2007,24(2):258-264.
[5] Jain RK,Au P,Tam J,et al.Engineering vascularized tissue[J].Nat Biotechnol,2005,23(7):821-823.
[6] Zisch AH,Lutolf MP,Hubbell JA.Biopolymeric delivery matrices for angiogenic growth factors[J].Cardiovasc Pathol,2003,12(6):295-310.
[7] 林诗晗,胡雪刚,吕红兵.人牙髓干细胞与根尖牙乳头干细胞miRNAs表达谱的差异分析[J].实用口腔医学杂志,2018,34(2):239-243.
[8] Schipani E,Maes C,Carmeliet G,et al.Regulation of osteogenesis- angiogenesis coupling by HIFs and VEGF[J].J Bone Miner Res,2009,24(8):1347-1353.
[9] Lin L,Shen Q,Leng H,et al.Synergistic inhibition of endochondral bone formation by silencing Hif1α and Runx2 in trauma- induced heterotopic ossification[J].Mol Ther,2011,19(8):1426-1432.
[10] 林成,刘宝林,刘彦普,等.VEGF和TGF- β1促进非血管化骨移植同期种植骨结合的实验研究[J].实用口腔医学杂志,2011,27(1):12-16.
[11] Ehrbar M,Metters A,Zammaretti P,et al.Endothelial cell proliferation and progenitor maturation by fibrin- bound VEGF variants with differential susceptibilities to local cellular activity[J].J Control Release,2005,101(1-3):93-109.
[12] Lian JB,Stein GS,van Wijnen AJ,et al.MicroRNA control of bone formation and homeostasis[J].Nat Rev Endocrinol,2012,8(4):212-227.
[13] Gallach S,Calabuig- Fariňas S,Jantus- Lewintre E,et al.MicroRNAs:Promising New Antiangiogenic Targets in Cancer[J].Biomed Res Int,2014,2014:878450.
[14] Boon RA,Seeger T,Heydt S,et al.MicroRNA- 29 in aortic dilation:Implications for aneurysm formation[J].Circ Res,2011,109(10):1115-1119.
[15] Wang S,Aurora AB,Johnson BA,et al.The endothelial- specific microRNA miR- 126 governs vascular integrity and angiogenesis[J].Dev Cell,2008,15(2):261-271.
[16] Fish JE,Santoro MM,Morton SU,et al.miR- 126 regulates angiogenic signaling and vascular integrity[J].Dev Cell,2008,15(2):272-284.
[17] Radom- Aizik S,Zaldivar FP Jr,Haddad F et al.Impact of brief exercise on circulating monocyte gene and microRNA expression:Implications for atherosclerotic vascular disease[J].Brain Behav Immun,2014,39:121-129.