| 关子易,刘萌萌,高义哲,等.X射线对血管内皮细胞GATA4表达及细胞衰老的影响[J].中华放射医学与防护杂志,2026,46(5):490-496.Guan Ziyi,Liu Mengmeng,Gao Yizhe,et al.Effects of X-ray irradiation on GATA4 expression and cellular premature senescence in human umbilical vein endothelial cells[J].Chin J Radiol Med Prot,2026,46(5):490-496 |
| X射线对血管内皮细胞GATA4表达及细胞衰老的影响 |
| Effects of X-ray irradiation on GATA4 expression and cellular premature senescence in human umbilical vein endothelial cells |
| 投稿时间：2025-05-08 |
| DOI：10.3760/cma.j.cn112271-20250508-00161 |
| 中文关键词: 电离辐射 内皮细胞 衰老 p62 GATA4 |
| 英文关键词:Ionizing radiation Endothelial cells Premature senescence p62 GATA4 |
| 基金项目:国家自然科学基金(31570852,82574030);北京市自然科学基金(7202139) |
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| 中文摘要: |
| 目的 探究不同剂量X射线对人脐静脉内皮细胞衰老的影响。方法 体外实验采用0.188、0.375、0.75、2、4 Gy X射线照射后收集人脐静脉内皮细胞EA.hy926。采用CCK-8试剂盒检测细胞增殖能力。使用β-半乳糖苷酶染色检测细胞衰老比例。采用Western blot方法检测蛋白p21、p16、p62、GATA4表达变化。采用实时荧光定量检测衰老相关分泌表型因子IL-33、IFIT-1、IFI-44 mRNA表达变化。体内实验采用2和4 Gy X射线对C57BL/6J小鼠全身照射,照后采用Western blot检测心脏组织p21、p62和GATA4表达变化。结果 照射后48 h,所有剂量组细胞增殖受抑制(t=6.07~17.10,P＜0.01);72 h时,低剂量(0.188和0.375 Gy)照射促进增殖(t=5.52、6.24,P＜0.05),高剂量(2和4 Gy)照射抑制增殖(t=4.18、3.39,P＜0.01)。2和4 Gy组β-半乳糖苷酶染色阳性细胞比例显著增加(t=12.34、11.49,P＜0.05)。0.75、2和4 Gy组p21蛋白表达升高(t=12.66、14.34、17.54,P＜0.05);2和4 Gy组p21和p16蛋白表达在照射后7 d显著增加(t=3.48、23.58,P＜0.05)。0.375、0.75、2和4 Gy照射后72 h,p62蛋白表达降低(t=9.88~51.12,P＜0.05),GATA4蛋白表达升高(t=7.10~25.91,P＜0.05)。2和4 Gy组小鼠心脏组织中p21蛋白表达增加(t=8.21、12.13,P＜0.05),p62/GATA4通路激活,p62蛋白减少(t=11.16、7.46,P＜0.05),GATA4蛋白增加(t=13.66、8.61,P＜0.05)。不同剂量照射后72 h,2、4 Gy组SASP因子IL-33 mRNA表达增加(t=28.98、21.17,P＜0.05);0.375~4 Gy组IFIT-1和IFI-44 mRNA表达增加(t=46.40~62.87、11.28~22.07,P＜0.05)。结论 X射线可诱导人脐静脉内皮细胞衰老,并激活p62/GATA4通路。 |
| 英文摘要: |
| Objective To investigate the effects of different doses of X-rays on senescence in human umbilical vein endothelial cells. Methods Human umbilical vein endothelial cells (EA.hy926) were collected after irradiation with 0.188, 0.375, 0.75, 2, and 4 Gy of X-rays. Cell proliferation was assessed using the CCK-8 assay kit. The proportion of senescent cells was determined via β-galactosidase staining. Western blot analysis was employed to detect changes in p21, p16, p62, and GATA4 protein expression. Real-time quantitative PCR measured mRNA expression levels of senescence-associated secretory phenotype factors IL-33, IFIT-1, and IFI-4. In vivo experiments involved whole-body irradiation of C57BL/6J mice with 2 and 4 Gy X-rays. Post-irradiation Western blot analysis assessed cardiac tissue expression of p21 and p62/GATA4. Results At 48 h after irradiation, cell proliferation was inhibited in all dose groups (t=6.07-17.10, P<0.01). At 72 h, low doses (0.188 and 0.375 Gy) promoted proliferation (t=5.52, 6.24, P<0.05), while high doses (2 and 4 Gy) inhibited proliferation (t=4.18, 3.39, P<0.01). The proportion of β-galactosidase-positive cells was significantly increased in the 2 and 4 Gy groups (t=12.34, 11.49, P<0.05). P21 protein expression was elevated in the 0.75, 2, and 4 Gy groups (t=12.66, 14.34, 17.54, P<0.05). In the 2 and 4 Gy groups, the expression of p21 and p16 proteins significantly increased 7 d after irradiation (t=3.48, 23.58, P<0.05). At 72 h after 0.375, 0.75, 2, and 4 Gy X-ray irradiation, p62 protein expression. decreased (t=9.88-51.12, P<0.05), while GATA4 protein expression increased (t=7.10-25.91, P<0.05). In the cardiac tissues of mice in the 2 and 4 Gy groups, p21 protein expression was increased (t=8.21, 12.13, P<0.05), the p62/GATA4 pathway was activated, with decreased p62 protein expression (t=11.16, 7.46, P<0.05) and increased GATA4 protein expression (t=13.66, 8.61, P<0.05). At 72 h after irradiation with different doses, mRNA expression of the SASP factor IL-33 increased in the 2 and 4 Gy groups (t=28.98, 21.17, P<0.05). In the 0.375-4 Gy groups, mRNA expression of IFIT-1 and IFI-44 increased (t=46.40-62.87, 11.28-22.07, P<0.05) Conclusions X-rays induce senescence in human umbilical vein endothelial cells and activate the p62/GATA4 pathway. |
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