体外培养大鼠心肌微血管内皮细胞缺氧模型的建立

发布时间：2019-05-19 20:28

【摘要】：目的建立大鼠心肌微血管内皮细胞(CMECs)缺氧模型。方法植块法培养CMECs,免疫细胞化学鉴定微血管内皮细胞特异性标志。将原代培养的CMECs在1%O2-94%N2-5%CO2低氧气体环境中培养24h制备低氧损伤模型(L组),以无血清常氧培养作为对照组(N组)。倒置相差显微镜观察缺氧前后细胞形态变化,MTT法测定细胞活力,Annexin V-FITC和PI双标记法测定细胞凋亡率。结果植块法培养的大鼠CMECs具备典型微血管内皮细胞特征;Ⅷ因子、CD31相关抗原免疫染色鉴定均阳性。缺氧前后细胞形态无明显变化;与N组比较,L组细胞活力明显增加(P0.05);细胞凋亡率明显降低(P0.05),其中早期凋亡降低最明显(P0.01)。结论本方法可以成功建立简便、易行的CMECs细胞缺氧模型。
[Abstract]:Objective to establish a rat model of (CMECs) hypoxia in myocardial microvascular endothelial cells. Methods the specific markers of microvascular endothelial cells were identified by CMECs, immunocytochemistry. The hypoxia injury model (group L) was established by culture of primary cultured CMECs in 1%O2-94%N2-5%CO2 anoxic gas environment for 24 h, and serum-free normoxic culture was used as control group (N group). The morphological changes of cells before and after hypoxia were observed by inverted phase contrast microscope, the cell vitality was measured by MTT assay, and the apoptosis rate was measured by Annexin V-FITC and PI double labeling. Results the rat CMECs cultured by block method had the characteristics of typical microvascular endothelial cells, and all of them were positive by immunostaining of factor VIII and CD31 related antigen. There was no significant change in cell morphology before and after hypoxia. Compared with group N, the activity of cells in group L was significantly increased (P 0.05), and the apoptosis rate was significantly decreased (P 0.05), especially in the early stage of apoptosis (P 0.01). Conclusion this method can successfully establish a simple and feasible hypoxia model of CMECs cells.
【作者单位】：山东中医药大学第一临床学院;山东中医药大学附属医院;
【基金】：国家自然科学基金资助项目(No.81173441)
【分类号】：R-332

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