摘要
目的研究血管紧张素Ⅱ(AngⅡ)诱导的人脐静脉内皮细胞(HUVECs)衰老及对白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)分泌的影响,进一步探讨AngⅡ在动脉粥样硬化(atherosclerosis,AS)发生、发展过程中所发挥的作用。方法体外培养HUVECs,采用CCK-8法检测细胞存活率,用Ang Ⅱ(终浓度10-6mol/L)干预,分为实验对照组、AngⅡ诱导组。以衰老相关β-半乳糖苷酶活性和细胞的增殖能力2种衰老标志物为主要观察指标,其中衰老相关β-半乳糖苷酶活性采用免疫化学染色方法,流式细胞术检测细胞周期来反映细胞的增殖能力;酶联免疫吸附法(ELISA)检测培养液中IL-6、TNF-α浓度。结果与对照组相比,AngⅡ诱导组存活的细胞数为对照组的(77.15±6.83)%;(81.80±0.92)%的细胞呈现β-半乳糖苷酶阳性染色,流式细胞仪检测细胞周期停滞于G0-G1[(89.4±3.4)%],证实细胞衰老;IL-6、TNF-α分泌增加(P均<0.01)。结论 AngⅡ可以诱导HUVECs衰老,其机制可能与IL-6、TNF-α分泌增加有关。
Objective To observe the release of interleukin-6 and tumor necrosis factor-α and the aging of senescent human umbilical vein endothelial cells (HUVECs) induced by angiotensin Ⅱ (AngⅡ). Methods HUVECs were cultured in vitro and the cell viability was observed by CCK-8.HUVECs were intervened by AngⅡand divided into two groups: the control group, AngⅡ group (stimulated with AngⅡ10-6 mol/L for 48 h). Two ageing markers of ageing associated β-galactosidase activity and cell proliferation activity were adopted as main indexes.β-galactosidase activity was evaluated by immunocytochemical method and cell proliferation activity was measured with flowcytometry. The levels of interleukin-6 and tumor necrosis factor-α in the cell culture were determined with ELISA. Results The cell viability by AngⅡ-induced cells was (77.15±6.83) %, the positive cell number of β-gal staining was significantly higher in AngⅡ-induced cells (81.80±0.92) % than that in the control cells,The cell cycle was at G0/G1(68.4±3.4)%,suggestting cell senescence. The levels of interleukin-6 and tumor necrosis factor-α were significantly elevated with the AngⅡ (P<0.01). Conclusions AngⅡ induced HUVECs senescence and one of vascular endothelial cell senescence mechanisms may be relevant to the up-regulated levels of interleukin-6 and tumor necrosis factor-α.
出处
《实用老年医学》
CAS
2013年第1期15-18,共4页
Practical Geriatrics
基金
广东省自然科学基金(8151001002000022)
广州市科技计划项目(2010GN-E00221)
