高表达整合素连接激酶对脐静脉内皮细胞的影响

发布时间：2018-05-16 15:34

本文选题：整合素连接激酶 + 脐静脉内皮细胞　；参考：《南京医科大学》2012年硕士论文

【摘要】：目的探讨高表达整合素连接激酶(Integrin-Linked Kinase, ILK)对体外培养人脐静脉内皮细胞(human umbilical vein endothelial cells, HUVECs)增殖及凋亡的影响。方法体外胰酶消化、培养人脐静脉内皮细胞；随机分为空病毒对照组与ILK病毒转染组。利用高表达整合素连接激酶的腺病毒转染脐静脉内皮细胞,诱导其在脐静脉内皮细胞中高表达。确定最佳的病毒感染复数。采用MTT法测定细胞增殖能力；流式细胞检测技术测定细胞增殖周期；EDU法测定内皮细胞DNA合成能力；流式细胞技术测定缺氧条件下内皮细胞的的凋亡情况,Western Blotting检测脐静脉内皮细胞的蛋白激酶B (Akt)、内皮一氧化氮合酶(eNOS)蛋白表达及其蛋白磷酸化水平。结果ILK在内皮细胞高表达后,MTT法、流式细胞法、EDU法检测结果均表明：整合素连接激酶病毒转染组细胞增殖能力明显高于空病毒转染对照组(P0.05或P0.01)。同时,ILK组细胞的凋亡率明显减少(P0.05)。 Western Blotting检测发现整合素连接激酶病毒转染组的蛋白激酶B、内皮一氧化氮合酶磷酸化水平较对照组明显升高(P0.05),而蛋白激酶B、内皮一氧化氮合酶的表达水平两组间无差异。结论整合素连接激酶能促进脐静脉内皮细胞的增殖抑制其凋亡,其机制可能通过激活下游Akt/eNOS通路。
[Abstract]:Objective to investigate the effect of integrin-linked kinase (ILK) expression on the proliferation and apoptosis of human umbilical vein endothelial cells (HUVECs) in vitro. Methods Human umbilical vein endothelial cells were cultured by trypsin digestion in vitro and randomly divided into empty virus control group and ILK virus transfection group. High expression of integrin-linked kinase was induced in umbilical vein endothelial cells (HUVECs) by transfection of adenovirus into umbilical vein endothelial cells (HUVEC). Determine the optimal number of viral infections. The proliferative ability of endothelial cells was measured by MTT and DNA synthesis by flow cytometry. Flow cytometry was used to detect the apoptosis of endothelial cells under hypoxia. The protein expression and protein phosphorylation of protein kinase B (PKB) and endothelial nitric oxide synthase (Enos) in umbilical vein endothelial cells (HUVEC) were detected by Western Blotting. Results after high expression of ILK in endothelial cells, the results of flow cytometry and flow cytometry showed that the proliferative ability of the cells transfected with integrin linked kinase virus was significantly higher than that of the control group (P0.05 or P0.01). At the same time, the apoptotic rate of ilk group decreased significantly (P 0.05). Western Blotting analysis showed that the levels of protein kinase B and endothelial nitric oxide synthase phosphorylation in the integrin linked kinase virus transfected group were significantly higher than those in the control group, but the expression level of protein kinase B and endothelial nitric oxide synthase had no difference between the two groups. Conclusion integrin-linked kinase can promote the proliferation of umbilical vein endothelial cells and inhibit its apoptosis, which may be mediated by activation of downstream Akt/eNOS pathway.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363

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