【摘要】：目的：探索溶血磷脂酰膽堿(Lysophosphatidylcholine, LPC)和質(zhì)子(H+)刺激人臍靜脈內(nèi)皮細(xì)胞(Human umbilical vein endothelial cell, HUVEC)上細(xì)胞質(zhì)子感知受體G2A后對(duì)細(xì)胞間粘附因子1(Intercellular adhesion molecule-1, ICAM-1)、血管細(xì)胞粘附因子1(Vascular cell adhesion molecule-1, VCAM-1)表達(dá)量的影響,研究相關(guān)受體介導(dǎo)信號(hào)通路,并探討動(dòng)脈硬化中的作用。 方法：培養(yǎng)HUVEC,短發(fā)夾RNA (short hairpin RNA, shRNA)-G2A質(zhì)粒轉(zhuǎn)染HUVEC,并用G418篩選穩(wěn)定表達(dá)細(xì)胞株。PCR檢測(cè)G2A在HUVEC細(xì)胞及干擾G2A基因的HUVEC細(xì)胞中的表達(dá)。在LPC和質(zhì)子作用下檢測(cè)HUVEC細(xì)胞及G2A基因干擾的HUVEC細(xì)胞中鈣離子濃度、ICAM-1和VCAM-1表達(dá)量；檢測(cè)Gi蛋白抑制劑百日咳毒素(pertussis toxin, PTX)在此過程中的作用。 結(jié)果：獲得了穩(wěn)定干擾的G2A基因的HUVEC細(xì)胞。質(zhì)子刺激HUVEC相比對(duì)照HUVECG2A表達(dá)量降低。LPC刺激升高HUVEC胞內(nèi)Ca2+濃度,質(zhì)子能降低由LPC動(dòng)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(xì)胞后ICAM-1, VCAM-1表達(dá)量比刺激未干擾細(xì)胞后ICAM-1, VCAM-1表達(dá)量降低。LPC單獨(dú)刺激G2A干擾細(xì)胞引起ICAM-1, VACM-1表達(dá)量下降；共同刺激G2A干擾細(xì)胞ICAM-1, VACM-1表達(dá)量無變化。 結(jié)論：質(zhì)子刺激能降低HUVEC中G2A表達(dá)。G2A是在HUVEC里中ICAM-1、VCAM-1表達(dá)主要相關(guān)受體。G2A干擾的HUVEC細(xì)胞中質(zhì)子拮抗LPC介導(dǎo)的ICAM-1、VCAM-1變化。LPC活化的G2A與Gi蛋白相偶聯(lián),繼而升高胞內(nèi)Ca2+濃度；質(zhì)子拮抗LPC引起的Ca2+濃度上升。研究結(jié)果提示,雙配體受體對(duì)動(dòng)脈硬化發(fā)生中起重要的調(diào)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(xué)位授予單位】：內(nèi)蒙古大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R363

【共引文獻(xiàn)】

相關(guān)期刊論文 前6條

1 程慧嫻;徐苗苗;高瑋;惠康麗;金孝\，

本文編號(hào)：2252098