本文選題：整合素連接激酶 + 臍靜脈內(nèi)皮細胞��； 參考：《南京醫(yī)科大學(xué)》2012年碩士論文

【摘要】：目的探討高表達整合素連接激酶(Integrin-Linked Kinase, ILK)對體外培養(yǎng)人臍靜脈內(nèi)皮細胞(human umbilical vein endothelial cells, HUVECs)增殖及凋亡的影響。 方法體外胰酶消化、培養(yǎng)人臍靜脈內(nèi)皮細胞；隨機分為空病毒對照組與ILK病毒轉(zhuǎn)染組。利用高表達整合素連接激酶的腺病毒轉(zhuǎn)染臍靜脈內(nèi)皮細胞,誘導(dǎo)其在臍靜脈內(nèi)皮細胞中高表達。確定最佳的病毒感染復(fù)數(shù)。采用MTT法測定細胞增殖能力；流式細胞檢測技術(shù)測定細胞增殖周期；EDU法測定內(nèi)皮細胞DNA合成能力；流式細胞技術(shù)測定缺氧條件下內(nèi)皮細胞的的凋亡情況,Western Blotting檢測臍靜脈內(nèi)皮細胞的蛋白激酶B (Akt)、內(nèi)皮一氧化氮合酶(eNOS)蛋白表達及其蛋白磷酸化水平。 結(jié)果ILK在內(nèi)皮細胞高表達后,MTT法、流式細胞法、EDU法檢測結(jié)果均表明：整合素連接激酶病毒轉(zhuǎn)染組細胞增殖能力明顯高于空病毒轉(zhuǎn)染對照組(P0.05或P0.01)。同時,ILK組細胞的凋亡率明顯減少(P0.05)。 Western Blotting檢測發(fā)現(xiàn)整合素連接激酶病毒轉(zhuǎn)染組的蛋白激酶B、內(nèi)皮一氧化氮合酶磷酸化水平較對照組明顯升高(P0.05),而蛋白激酶B、內(nèi)皮一氧化氮合酶的表達水平兩組間無差異。 結(jié)論整合素連接激酶能促進臍靜脈內(nèi)皮細胞的增殖抑制其凋亡,其機制可能通過激活下游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.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R363

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