论文部分内容阅读
目的观察不同浓度血管紧张素Ⅱ(AngⅡ)对诱导人脐静脉血管内皮细胞(HUVEC)衰老和P-选择素表达的影响。方法体外培养HUVEC,随机分为对照组和含不同浓度AngⅡ组(10-8～10-5mmol/L)共5组,用含不同浓度AngⅡ的培养基诱导细胞衰老,β-半乳糖苷酶(β-gal)染色法鉴定内皮细胞衰老状态;流式细胞技术分析细胞周期变化;CCK-8法检测细胞存活率情况;ELISA检测各组培养基上清中P-选择素表达含量。结果 AngⅡ培养细胞48h后,β-gal阳性染色率随着AngⅡ浓度的增加逐渐增多(P均<0.01);细胞周期多停滞于G0/G1期,S期细胞逐渐减少,与对照组相比,10-8和10-7mmol/LAngⅡ组无明显差异(P>0.05),10-6和10-5mmol/LAngⅡ组有统计学差异(P<0.05或P<0.01);细胞存活率与对照组相比明显下降;P-选择素表达含量随着AngⅡ浓度的增加明显增多(P均<0.01)。结论 AngⅡ诱导HUVEC衰老,并呈剂量依赖性的抑制细胞增殖与增加P-选择素的表达。
Objective To observe the effects of different concentrations of angiotensin Ⅱ on the senescence and P-selectin expression in human umbilical vein endothelial cells (HUVECs). Methods HUVECs were cultured in vitro and randomly divided into control group and AngⅡ group (10-8 ~ 10-5mmol / L) for 5 days. Cell senescence was induced by culture medium containing different concentrations of AngⅡ, and β-galactosidase β-gal staining was used to identify the aging status of endothelial cells. Flow cytometry was used to analyze cell cycle changes. CCK-8 was used to determine cell viability. ELISA was used to detect the expression of P-selectin in supernatant of each group. Results Compared with the control group, the positive staining rate of β-gal increased gradually with the increase of AngⅡ concentration (P <0.01) after 48 hours of AngⅡ culture. The cell cycle was arrested in G0 / G1 phase and the number of S phase cells was decreased. There was no significant difference between 10-8 and 10-7mmol / LAngⅡgroup (P> 0.05), while the difference between 10-6 and 10-5mmol / LAngⅡgroup was statistically significant (P <0.05 or P <0.01) P-selectin expression increased significantly with the increase of AngⅡ concentration (all P <0.01). Conclusion AngⅡ induced the senescence of HUVECs and inhibited the proliferation and increased the expression of P-selectin in a dose-dependent manner.