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目的 观察在大剂量放射损伤条件下 ,内毒素活化的U93 7对脐静脉内皮细胞ECV3 0 4分泌的细胞因子及其生物学性状的作用。方法 将ECV3 0 4未照射组设为对照组 ,ECV3 0 4+12Gy60 Co照射组和ECV3 0 4+U93 7+12Gy60 Co照射组设为实验组 ,照射后培养 48h ,测定细胞的存活率、细胞周期及凋亡方面的变化 ,运用ELISA法测定培养上清中的TGF -β1和VEGF的浓度变化 ,运用免疫荧光 +流式细胞技术检测细胞表面血管内皮生长因子受体 2表达水平 ,运用逆转录聚合酶链反应检测细胞KDR的mRNA水平。结果 大剂量辐射引起细胞增殖率降低 ,凋亡增加 ,G2 /M期阻滞 ,促进ECV3 0 4分泌VEGF和TGF -β1,提高了KDR的表达 ,巨噬细胞可以缓解细胞增殖率的降低 ,减少细胞凋亡和G2 /M期阻滞 ,进一步提高VEGF的分泌和KDR的表达 ,减少TGF -β1的分泌。结论 放射引起细胞增殖率降低 ,凋亡增加 ,G2 /M期阻滞 ,部分与细胞加强分泌的TGF -β1的抑制作用有关 ;在放射损伤的情况下 ,巨噬细胞对内皮细胞是起着积极的保护性作用的
Objective To observe the effect of endotoxin-activated U93 7 on cytokines secreted by human umbilical vein endothelial cells (ECV304) and their biological properties under high-dose radiation injury. Methods The ECV3 0 4 non-irradiated group was set as the control group. The ECV3 0 4 + 12Gy60 Co irradiation group and the ECV3 0 4 + U93 7 + 12Gy60 Co irradiation group were set as the experimental group. After irradiation for 48 hours, the cell viability, The changes of TGF-β1 and VEGF in culture supernatant were measured by ELISA. The expression of vascular endothelial growth factor receptor 2 (VEGFR2) was detected by immunofluorescence and flow cytometry. Polymerase chain reaction was used to detect KDR mRNA levels in cells. Results High-dose radiation caused a decrease of cell proliferation rate, increased apoptosis and G2 / M arrest, promoted the secretion of VEGF and TGF-β1 by ECV304 and increased the expression of KDR. Macrophages could alleviate the decrease of cell proliferation and decrease Apoptosis and G2 / M phase arrest further increased the secretion of VEGF and the expression of KDR, and decreased the secretion of TGF-β1. CONCLUSION: Radiation can reduce the cell proliferation rate, increase the apoptosis rate and arrest the G2 / M phase, which is partially related to the inhibitory effect of TGF-β1 secreted by the cells. In the case of radiation injury, the macrophages play a positive role in the endothelial cells Protective effect