EUK-207 通过 Bcl2 通路保护人肠道微血管内皮细胞（HIMEC）免受辐射诱导的细胞凋亡。
EUK-207 protects human intestinal microvascular endothelial cells (HIMEC) against irradiation-induced apoptosis through the Bcl2 pathway.
机构信息
Department of Surgery, Medical College of Wisconsin, Milwaukee, WI 53226, United States.
出版信息
Life Sci. 2012 Oct 22;91(15-16):771-82. doi: 10.1016/j.lfs.2012.08.018. Epub 2012 Aug 23.
AIM
To elucidate the signaling mechanisms involved in the protective effect of EUK-207 against irradiation-induced cellular damage and apoptosis in human intestinal microvasculature endothelial cells (HIMEC).
METHODS
HIMECs were irradiated and treated with EUK-207. Using hydroethidine and DCF-DA fluorescent probe the intracellular superoxide and reactive oxygen species (ROS) were determined. By real-time PCR and western blotting caspase-3, Bcl2 and Bax genes and proteins were analyzed. Proliferation was determined by [(3)H]-thymidine uptake. Immunofluorescence staining was used for translocation of p65 NFκB subunit.
KEY FINDING
Irradiation increased ROS production, apoptosis, Bax, Caspase3 and NFkB activity in HIMEC and inhibited cell survival/growth/proliferation. EUK-207 restored the endothelial functions, markedly inhibited the ROS, up-regulated the Bcl2 and down-regulated Bax and prevented NFκB caspase 3 activity in HIMEC.
SIGNIFICANCE
HIMEC provide a novel model to define the effect of irradiation induced endothelial dysfunction. Our findings suggest that EUK-207 effectively inhibits the damaging effect of irradiation.
目的
阐明 EUK-207 对人肠道微血管内皮细胞（HIMEC）辐射诱导细胞损伤和凋亡的保护作用涉及的信号机制。
方法
用 EUK-207 处理和辐照 HIMEC。使用羟乙基二氢二嗪和 DCF-DA 荧光探针测定细胞内超氧阴离子和活性氧（ROS）。通过实时 PCR 和 Western blot 分析 caspase-3、Bcl2 和 Bax 基因和蛋白。通过 [(3)H]-胸苷摄取测定增殖。免疫荧光染色用于检测 p65 NFκB 亚基的易位。
主要发现
辐照增加了 HIMEC 中的 ROS 产生、细胞凋亡、Bax、Caspase3 和 NFkB 活性，并抑制了细胞存活/生长/增殖。EUK-207 恢复了内皮功能，显著抑制了 ROS，上调了 Bcl2，并下调了 Bax，防止了 HIMEC 中的 NFκB caspase 3 活性。
意义
HIMEC 提供了一个新的模型来定义辐射诱导的内皮功能障碍的影响。我们的研究结果表明，EUK-207 能有效抑制辐射的损伤作用。
Zotero 插件