CAJ | 학술논문

目的：探讨缺氧下大鼠心肌微血管内皮细胞（CMECs）血管新生能力的变化。方法将原代培养的大鼠CMECs在1％O2、94％N2、5％CO2低氧气体环境中培养24 h制备缺氧损伤模型（缺氧组），以无血清常氧培养CMECs作为对照组。采用划痕愈合试验检测细胞迁移能力；倒置相差显微镜计数细胞成管情况；噻唑蓝比色法绘制细胞生长曲线并检测细胞增殖率；动态观察寻找细胞迁移、成管、增殖的窗口期。 Real-time PCR法检测细胞增殖相关蛋白激酶（ERK）和细胞凋亡相关分子（p53）mRNA的表达。结果缺氧组细胞增殖率、迁移率和成管率均低于对照组，P均＜0．05。与对照组比较，缺氧组血管新生的顺序发生了改变，增殖的窗口期提前至第2天，与成管窗口期重叠。缺氧组、对照组ERK mRNA相对表达量分别为1．681±0．403、1．000，p53 mRNA相对表达量分别为0．458±0．100、1．000，缺氧组ERK mRNA表达高于对照组，p53 mRNA表达低于对照组（P均＜0．05）。结论缺氧24 h后大鼠CMECs血管新生能力明显降低，可能与p53基因表达下调、ERK基因表达上调有关。
목적：탐토결양하대서심기미혈관내피세포（CMECs）혈관신생능력적변화。방법장원대배양적대서CMECs재1％O2、94％N2、5％CO2저양기체배경중배양24 h제비결양손상모형（결양조），이무혈청상양배양CMECs작위대조조。채용화흔유합시험검측세포천이능력；도치상차현미경계수세포성관정황；새서람비색법회제세포생장곡선병검측세포증식솔；동태관찰심조세포천이、성관、증식적창구기。 Real-time PCR법검측세포증식상관단백격매（ERK）화세포조망상관분자（p53）mRNA적표체。결과결양조세포증식솔、천이솔화성관솔균저우대조조，P균＜0．05。여대조조비교，결양조혈관신생적순서발생료개변，증식적창구기제전지제2천，여성관창구기중첩。결양조、대조조ERK mRNA상대표체량분별위1．681±0．403、1．000，p53 mRNA상대표체량분별위0．458±0．100、1．000，결양조ERK mRNA표체고우대조조，p53 mRNA표체저우대조조（P균＜0．05）。결론결양24 h후대서CMECs혈관신생능력명현강저，가능여p53기인표체하조、ERK기인표체상조유관。
Objective To investigate the angiogenesis ability changes of rat cardiac microvascular endothelial cells ( CMECs) under hypoxic conditions .Methods The primarily cultured rat CMECs were cultured under hypoxia of 1%O2 , 94%N2 , and 5%CO2 for 24 h to prepare hypoxic injury models ( hypoxia group ) , meanwhile , we set the control group which was cultured under normoxic serum-free condition .Scratch healing assay was used to detect the cell migration , in-verted phase-contrast microscope was employed to count cell tube formation , MTT colorimetric assay was used to draw the cell growth curve and detect the cell proliferation rate , and dynamic observation method was applied to find the window pe-riods of cell migration , tube formation and proliferation .Real-time PCR was used to detect the expression of cell prolifera-tion-associated protein kinase (ERK) and apoptosis-related molecules (p53).Results The cell proliferation rate, migra-tion rate and tube formation rate of the hypoxia group were significantly lower than those of the control group ( all P<0.05).Compared with the control group , the angiogenesis order of the hypoxia group changed , the window period of prolif-eration became the second day in advance which was the same day of the tube formation window .The ERK mRNA relative expression levels of the hypoxia and control groups were 1.681 ±0.403 and 1.000, and p53 mRNA relative expression lev-els were 0.458 ±0.100 and 1.000.ERK mRNA expression of the hypoxia group was higher , p53 mRNA expression was lower than that of the control group (all P<0.05 ).Conclusion The angiogenesis abilities of rat CMECs significantly re-duce under hypoxic environment for 24 h, which may be related to the down-regulation of p53 gene and up-regulation of ERK gene.