摘要
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 。
Objective To investigate the effect of strong ion-irradiation and active macrophage on vascular endothelial cells. Methods The vascular endothelial cells(ECV-304) were divided into 3 groups: ECV304(A), ECV-304 with radiation (B)and ECV-304 with active macrophage U937 and radiation(C). The survival rates, cell cycle and apoptosis of ECV-304 were determined 48 hours after irradiation. The concentrations of TGF-β1 and VEGF in the cultured supernatant were determined by ELISA method.The protein expression of the endothelial receptor KDR in ECV-304 cells was detected by immunofluorescence and flow-cytometry while its mRNA expression was detected by RT-PCR. Results After high dose of ion-irrdiation, the cellular proliferation activity decreased, apopototic cells increased and the cell ratio in G2/M phase increased.In addition,The secretion of VEGF and TGF-β1 was promoted with the increased KDR expression. Active macrophages could alleviate the decrease of cellular proliferation and the increase of apoptotic cells and cell ratio in G2/M phase induced by ion-irriation.Furthermore,the VEGF secretion and the KDR expression were enhanced and the secretion of TGF-β1 was inhibited by the active macrophages. Conclusions Ion irradiation could induce the decrease of cellular proloferation, the increase of apoptotic cells and the cells in G2/M phase.This might partly be related to the inhibiting roles of TGF-β1 secreted by the ECV-304 cells.Active macrophages might protect the endothelial cells from radiation injury.
出处
《中国医师杂志》
CAS
2003年第11期1456-1459,共4页
Journal of Chinese Physician
