紫绀及非紫绀型先心病患儿来源的hMSCs促血管生成能力比较  

Comparison of pro-angiogenetic ability of hMSCs from children with cyanotic and acyanotic congenital heart disease

作　　者：蔡俊 赵俊涛[1] 张伟峰[1] CAI Jun;ZHAO Jun-tao;ZHANG Wei-feng(the 7th People's Hospital of Zhengzhou, Zhengzhou 450000, China)

机构地区：[1]郑州市第七人民医院

出　　处：《临床医学研究与实践》2019年第25期3-5,共3页Clinical Research and Practice

摘　　要：目的比较紫绀型先天性心脏病(C-CHD)患儿和非紫绀型先天性心脏病(A-CHD)患儿来源的骨髓干细胞(hMSCs)在乏氧环境下的促血管生成能力,并探讨其发生机制。方法 hMSCs取自C-CHD患儿(C组)和A-CHD患儿(A组),体外于乏氧环境下(1%O2,5%CO2,94%N2)培养。以CCK8法比较两组细胞的增殖能力;以结晶紫法比较两组细胞的克隆形成能力;以Western Blot法检测两组细胞中血管内皮细胞生长因子A(VEGFA)及其受体(VEGFR)的蛋白表达水平;乏氧无血清条件下,以ELISA法检测两组细胞分泌VEGFA的情况。结果在乏氧环境下,相比于A组,C组细胞具有更强的细胞增殖能力、更强的克隆形成能力(P<0.05);C组细胞中的VEGFA、VEGFR蛋白表达水平明显高于A组(P<0.05);在无氧无血清的应激情况下,相比于A组,C组细胞个体可分泌更多的VEGFA(P<0.05)。结论 C-CHD患儿来源的hMSCs具有更好的促血管生成能力,这可能与其天然乏氧诱导细胞体内VEGFA合成增多,VEGFR表达上调,乏氧应激情况下可分泌更多的VEGFA有关。Objective To compare the pro-angiogenitic ability of human mesenchymal stem cells (h-MSCs) from children with cyanotic congenital heart disease (C-CHD) and acyanotic congenital heart disease (A-CHD) in hypoxic environment, and explore its mechanism. Methods The h-MSCs were isolated from children with C-CHD (group C) and A-CHD (group A) and cultured in hypoxic environment in vitro (1% O2, 5% CO2, 94% N2). CCK8 assay was used to compare the proliferation ability of cells in the two groups;crystal violet method was used to compare the clonogenic ability of cells in the two groups;Western Blot assay was used to detect the protein expression levels of vascular endothelial growth factor A (VEGFA) and its receptor (VEGFR) of cells in the two groups;under hypoxic and serum-free conditions, the secretion of VEGFA was detected by ELISA. Results In hypoxic environment, compared with the group A, the group C had stronger cell proliferation and cloning formation ability (P<0.05). The protein expression levels of VEGFA and VEGFR in the group C were significantly higher than those in the group A (P<0.05). Under the condition of anaerobic and serum-free stress, compared with the group A, individual cells in the group C secreted more VEGFA (P<0.05). Conclusion C-CHD-derived hMSCs presented the superiority for pro-angiogenesis, and the possible mechanism was the up-regulation of VEGF and VEGFR, and the enhanced secretion of VEGF under hypoxic stress.

关 键 词：紫绀 骨髓干细胞 先天性心脏病 血管化 

分 类 号：R725.4[医药卫生—儿科]