| 李辰,田梅,苟巧,刘建香,齐雪松,王春燕,苏旭.缝隙连接蛋白43在X射线诱导的人脐静脉内皮细胞凋亡中的作用及机制研究[J].中华放射医学与防护杂志,2018,38(10):728-733 |
| 缝隙连接蛋白43在X射线诱导的人脐静脉内皮细胞凋亡中的作用及机制研究 |
| Role of connexin 43 in X-ray induced apoptosis in human umbilical vein endothelial cells and its mechanism |
| 投稿时间：2018-08-28 |
| DOI：10.3760/cma.j.issn.0254-5098.2018.10.002 |
| 中文关键词: X射线 HUVEC细胞 Cx43 凋亡 cleaved caspase-3 |
| 英文关键词:X-rays HUVEC cell Cx43 Apoptosis cleaved caspase-3 |
| 基金项目:科研院所社会公益研究专项（2005DIB1J087） |
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| 中文摘要: |
| 目的 研究缝隙连接蛋白43（Cx43）在X射线致人脐静脉内皮细胞（HUVEC）凋亡中的作用并探讨其机制。方法 采用流式细胞术检测10 Gy X射线照射后48~96 h以及不同剂量X射线照射后72 h HUVEC细胞凋亡的变化。Western blot检测0、5、10、20 Gy X射线照射后72 h HUVEC中Cx43和cleaved caspase-3蛋白的表达。采用RNA干扰技术使细胞Cx43表达沉默，或转染Cx43高表达质粒使Cx43过表达。流式细胞术和Western blot分别检测Cx43沉默和过表达对受照HUVEC凋亡的影响以及cleaved caspase-3蛋白表达水平的变化。结果 X射线照射后48~96 h HUVEC凋亡增加并且呈现剂量依赖性。在0~20 Gy范围内，Cx43表达随剂量增加而降低，cleaved caspase-3表达随剂量增加而增高。Cx43沉默组HUVEC细胞早期凋亡及凋亡死亡细胞比例明显高于无意义序列组（t=3.674、6.375，P<0.05）；Cx43过表达组HUVEC细胞早期凋亡及凋亡死亡细胞比例明显低于空载体组（t=9.399、11.190，P<0.05）；Cx43沉默组较无意义序列组cleaved caspase-3表达增强，而过表达组低于空载体组。结论 Cx43通过调控cleaved caspase-3活化，抑制HUVEC细胞凋亡，从而对X射线照射的HUVEC细胞发挥保护作用。 |
| 英文摘要: |
| Objective To study the role of Cx43 in X-ray induced apoptosis of HUVEC cells and its mechanism. Methods Flow cytometry was used to detect the apoptosis of HUVEC cells at 48-96 h after 10 Gy X-ray irradiation and at 72 h after irradiation of different doses. Western blot was used to detect the protein expressions of Cx43 and cleaved caspase-3 in HUVEC cells at 72 h after 0, 5, 10 and 20 Gy irradiation. Small interfering RNA was transfected into HUVEC cells to silence Cx43 expression, the Cx43 bearing plasmid was transfected into cells to overexpress Cx43. The effect of Cx43 knockdown or overexpression on apoptosis induction and cleaved caspase-3 protein expression were detected by flow cytometry and Western blot, respectively. Results The apoptosis of HUVEC increased significantly from 48 h to 96 h after X-ray irradiation and in a dose-dependent manner at 72 h after irradiation. The expression of Cx43 protein was negatively correlated with the dose but the expression of cleaved caspase-3 was positively correlated with the dose in the range of 0-20 Gy. After Cx43 silencing, the proportion of early apoptosis and apoptosis combined with dead cells were significantly higher than that of the siRNA control group(t=3.674, 6.375, P<0.05). After Cx43 overexpression, the proportion of early apoptosis and apoptosis combined with dead cells were significantly lower than that of vector control group(t=9.399, 11.190, P<0.05). The expression of cleaved caspase-3 in the Cx43 silencing group was higher than that in the siRNA control group, but this protein in the Cx43 overexpressed group was lower than that in the vector control group. Conclusions Cx43 may protect X-ray irradiated HUVEC cells from apoptosis by down-regulating the activation of caspase-3. |
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