【摘要】：目的：探索溶血磷脂酰膽堿(Lysophosphatidylcholine, LPC)和質子(H+)刺激人臍靜脈內皮細胞(Human umbilical vein endothelial cell, HUVEC)上細胞質子感知受體G2A后對細胞間粘附因子1(Intercellular adhesion molecule-1, ICAM-1)、血管細胞粘附因子1(Vascular cell adhesion molecule-1, VCAM-1)表達量的影響,研究相關受體介導信號通路,并探討動脈硬化中的作用。 方法：培養(yǎng)HUVEC,短發(fā)夾RNA (short hairpin RNA, shRNA)-G2A質粒轉染HUVEC,并用G418篩選穩(wěn)定表達細胞株。PCR檢測G2A在HUVEC細胞及干擾G2A基因的HUVEC細胞中的表達。在LPC和質子作用下檢測HUVEC細胞及G2A基因干擾的HUVEC細胞中鈣離子濃度、ICAM-1和VCAM-1表達量；檢測Gi蛋白抑制劑百日咳毒素(pertussis toxin, PTX)在此過程中的作用。 結果：獲得了穩(wěn)定干擾的G2A基因的HUVEC細胞。質子刺激HUVEC相比對照HUVECG2A表達量降低。LPC刺激升高HUVEC胞內Ca2+濃度,質子能降低由LPC動員的Ca2+濃度。PTX能抑制由LPC引起的Ca2+濃度上升。LPC刺激G2A干擾細胞后ICAM-1, VCAM-1表達量比刺激未干擾細胞后ICAM-1, VCAM-1表達量降低。LPC單獨刺激G2A干擾細胞引起ICAM-1, VACM-1表達量下降；共同刺激G2A干擾細胞ICAM-1, VACM-1表達量無變化。 結論：質子刺激能降低HUVEC中G2A表達。G2A是在HUVEC里中ICAM-1、VCAM-1表達主要相關受體。G2A干擾的HUVEC細胞中質子拮抗LPC介導的ICAM-1、VCAM-1變化。LPC活化的G2A與Gi蛋白相偶聯,繼而升高胞內Ca2+濃度；質子拮抗LPC引起的Ca2+濃度上升。研究結果提示,雙配體受體對動脈硬化發(fā)生中起重要的調控作用。
[Abstract]:Objective: to investigate the effects of lysophosphatidylcholine (Lysophosphatidylcholine, LPC) and proton (H) (H) on cytoplasmic adhesion factor 1 (Intercellular adhesion molecule-1, ICAM-1) and vascular cell adhesion factor 1 (Vascular cell adhesion molecule-1, (Vascular cell adhesion molecule-1,) on human umbilical vein endothelial cells (Human umbilical vein endothelial cell, HUVEC). The effect of VCAM-1 expression, To study the signal pathway mediated by related receptors and to explore the role of atherosclerosis. Methods: the expression of G2A in HUVEC cells and HUVEC cells which interfered with G2A gene was detected by G418 screening and stable expression cell line. The expression of ICAM-1 and VCAM-1 in HUVEC cells and G2A gene interfering HUVEC cells were detected by LPC and proton, and the role of Gi protein inhibitor pertussis toxin (pertussis toxin, PTX) in this process was detected. Results: stable interfering G 2A gene HUVEC cells were obtained. Proton stimulated HUVEC decreased compared with control HUVECG2A expression. LPC-stimulated increased intracellular Ca2 concentration of HUVEC. Proton could reduce the concentration of Ca2 mobilized by LPC. PTX could inhibit the increase of Ca2 concentration induced by LPC. The expression of ICAM-1, VCAM-1 in G2A interfering cells stimulated by LPC was lower than that of ICAM-1, VCAM-1 after stimulation of undisturbed cells. Co-stimulation of G 2A interfered with the expression of ICAM-1, VACM-1 in the cells. Conclusion: proton stimulation can reduce the expression of G2A. G2A in HUVEC. G2A is a major receptor. G2A interferes with the expression of G2A in HUVEC. Proton antagonizes ICAM-1,VCAM-1 changes mediated by LPC. G2A activated by LPC is coupled with Gi protein, and then increases the concentration of Ca2. Proton antagonizes the increase of Ca2 concentration induced by LPC. The results suggest that double ligand receptors play an important role in the pathogenesis of atherosclerosis.
【學位授予單位】：內蒙古大學
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R363

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1 程慧嫻;徐苗苗;高瑋;惠康麗;金孝\，

本文編號：2252098