Caveolin-1在电离辐射诱导血管内皮细胞早衰中的作用研究  

Study on the role of Caveolin-1 in ionizing radiation-induced premature senescence of vascular endothelial cells

作　　者：伊如罕 陆雪[1] 蔡恬静[1] 高玲[1] Yi Ruhan;Lu Xue;Cai Tianjing;Gao Ling(Key Laboratory of Radiological Protection and Nuclear Emergency,China CDC,National Institute for Radiological Protection,Chinese Center for Disease Control and Prevention,Beijing 100088,China)

机构地区：[1]中国疾病预防控制中心辐射防护与核安全医学所辐射防护与核应急中国疾病预防控制中心重点实验室,北京100088

出　　处：《中华放射医学与防护杂志》2025年第3期163-169,共7页Chinese Journal of Radiological Medicine and Protection

摘　　要：目的探讨Caveolin-1(CAV-1)在电离辐射诱导血管内皮细胞早衰中的作用机制。方法在人微血管内皮细胞(HMEC-1)中,通过慢病毒转染方法,利用嘌呤霉素筛选构建稳定敲低CAV-1的细胞模型,根据是否感染慢病毒shRNA-CAV-1将细胞分为对照(NC)组和sh-CAV-1组。采用Western blot检测CAV-1蛋白表达水平。0、2、4 Gy X射线照射人微血管内皮细胞后24、48和72 h,利用β-半乳糖苷酶染色试剂盒检测细胞β-半乳糖苷酶染色情况;利用Western blot检测衰老标志蛋白p53蛋白和p21蛋白表达情况;用CCK-8试剂盒检测细胞活力变化;通过血管生成实验检测血管内皮细胞功能。结果2、4 Gy X射线照射后48 h,CAV-1蛋白表达显著下降(t=3.50、3.89,P<0.05)。0、2、4 Gy X射线照射后,与NC组相比,sh-CAV-1组β-半乳糖苷酶染色显著增加(t=12.91、11.54、6.04,P<0.05);在HMEC-1细胞中敲低CAV-1,细胞衰老相关蛋白p53表达水平显著下降(t=4.09、3.13、3.43,P<0.05),p21蛋白表达水平显著增加(t=-3.63、-3.33、-3.06,P<0.05)。与NC组相比,CAV-1敲低的HMEC-1细胞活力显著下降(t=2.97~25.89,P<0.05);血管生成能力降低(t=3.39~39.68,P<0.05)。结论CAV-1通过正向调控p53,负调控p21参与电离辐射诱导血管内皮细胞早衰发生过程。Objective To explore the role of Caveolin-1(CAV-1)in radiation-induced premature senescence of vascular endothelial cells.Methods A cell model with stable knockdown of CAV-1 was constructed in human microvascular endothelial cells(HMEC-1)by lentiviral transfection using puromycin screening.The cells were divided into NC group and sh-CAV-1 group based on whether they were infected with lentivirus shRNA-CAV-1.The protein expression levels of CAV-1,p53 and p21 were detected by Western blot at 24,48,and 72 h after 0,2,and 4 Gy X-ray irradiation.Theβ-galactosidase staining kit was used to detectβ-galactosidase in cells.CCK-8 kit was used to detect cell viability,and vascular endothelial cell function was detected by vascular tube-forming assay.Results CAV-1 protein expression was significantly decreased at 48 h after 2 and 4 Gy X-ray irradiation(t=3.50,3.89,P<0.05),andβ-galactosidase in sh-CAV-1 group was significantly increased at 72 h after 0,2 and 4 Gy X-ray irradiation(t=12.91,11.54,6.04,P<0.05)compared with the NC group.Knockdown of CAV-1 resulted in the decrease in the expression level of the cellular senescence-associated protein p53 protein(t=4.09,3.13,3.43,P<0.05),but increase in the expression level of p21 protein(t=-3.63,-3.33,-3.06,P<0.05).Compared with the NC group,knockdown CAV-1 significantly decreased cell viability(t=2.97-25.89,P<0.05)and reduced vessel-forming capacity(t=3.39-39.68,P<0.05).Conclusions CAV-1 is involved in the process of radiation-induced premature senescence of vascular endothelial cells through positive regulation of p53 and negative regulation of p21.

关 键 词：电离辐射 X射线 血管内皮细胞 早衰 CAVEOLIN-1 

分 类 号：R73[医药卫生—肿瘤]