塞来昔布对人脑微血管内皮细胞放射后凋亡及释放PGI2和TXA2影响  

The effects of Celecoxib on human brain microvascular endothelial cells release 6-keto-PGF1α and TXB2 and apoptosis after radiation

作　　者：孙佳星[1] 徐晓婷[1] 涂或[2] 

机构地区：[1]苏州大学附属第一医院放疗科,215006 [2]苏州大学医学部放射医学与防护学院,215123

出　　处：《中华放射肿瘤学杂志》2017年第6期682-686,共5页Chinese Journal of Radiation Oncology

基　　金：国家自然科学基金（81302384）

摘　　要：目的 探讨塞来昔布对人脑微血管内皮细胞（HBMECs）放射后释放PGI2、TXA2及凋亡影响。方法 细胞按空白、接受X射线、塞来昔布联合X射线分为对照组、单纯照射组、联合组,采用CCK-8、克隆形成实验测定塞来昔布对HBMECs毒性及放射敏感性影响。于照后6、12、24、48 h时间点进行实验。ELISA检测培养液6-keto-PGF1α、TXB2含量及计算TXB2/6-keto-PGF1α比值,AnnexinV-FITC/PI双染法测定细胞凋亡率,蛋白印迹法检测蛋白表达情况。配对t检验差异。结果30 μmol/L塞来昔布浓度下联合组未见明显放射增敏效应（SER=0.96）;与对照组相比,两组TXB2/6-keto-PGF1α比值升高（P〈0.05）,凋亡率升高（P〈0.05）,Cox-2、P-JNK、Cleaved caspase-3表达增加;与单纯放射组比较,联合组TXB2/6-keto-PGF1α比值降低（P〈0.05）,凋亡率降低（P〈0.05）,Cox-2及P-JNK、Cleaved caspase-3表达降低。结论 塞来昔布能减少放射后HBMECs释放TXB2,降低TXB2/6-keto-PGF1α比值,抑制细胞凋亡,抗凋亡机制与抑制Cox-2及P-JNK、Cleaved caspase-3表达有关。Objective To investigate the effect of Celecoxib on human brain microvascular endothelial cells release6-keto-PGF1α,TXB2 and apotosis after irradiation. Methods The logarithmic growth phase cells were divided into control groups （Con）,simple irradiation （IR） groups and combination groups （IR＋C）.CCK-8 and clone formation experiment were used to evaluate the effects of radiosensitivity and toxicity of celecoxib. The results were observed atthe time point of 6 h,12 h,24 h,48 h after irradiation. ELISA was used to test the contents of 6-keto-PGF1α and TXB2,which metabolized by PGI2 and TXA2 from culture medium after irradiation at different time points in different groups. TXB2/6-keto-PGF1αratios were calculated. Annexin V-FITC/PI double staining method was used to measure the apoptosis rates at different time points in different groups. Western blot was used to measure the protein expression. Paired t test difference. Results Compared with simple irradiation group,there were no significant radiosensitivity （SER=0.96） in combination groups incubated with30 μmol/L of celecoxib. Compared with the control group,the ratio of TXB2/6-keto-PGF1αincreased at each time point in IR and IR＋C （P〈0.05）,and the apoptosis rates increased （P〈0.05）.Cox-2,P-JNK and Cleaved caspase-3 increased. Compared with IR,the ratio of TXB2/6-keto-PGF1αdecreased at each time point in IR＋C （P〈0.05）,and the apoptosis rates decreased （t=3.34~6.38,P〈 0.05）.The protein expression of Cox-2,P-JNK and Cleaved caspase-3 decreased. Conclusions Celecoxib may help to protect HBMECs from releasing TXA2 and decreasing the ratio of TXB2/6-keto-PGF1α,and inhibitting apoptosis after irradiation. The mechanisms of apoptosis inhibition may be related to the inhibition of Cox-2 and P-JNK,caspase-3 Cleaved proteinexpressions.

关 键 词：人脑微血管内皮细胞 环氧合酶 塞来昔布 放射 前列环素 血栓素 凋亡 

分 类 号：R730.55[医药卫生—肿瘤]