The effects of Celecoxib on human brain microvascular endothelial cells release 6-keto-PGF1α and TXB2 and apoptosis after radiation
Sun Jiaxing,Xu Xiaoting,Tu Yu
Department of Radiation Oncology,The First Affiliated Hosptial of Soochow University,Suzhou 215006,China (Sun JX,Xu XT);School of Radiation Medicine and Protection,Medical School University,suzhou 215123,China (Tu Y)
Abstract： 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.
Sun Jiaxing,Xu Xiaoting,Tu Yu. The effects of Celecoxib on human brain microvascular endothelial cells release 6-keto-PGF1α and TXB2 and apoptosis after radiation[J]. Chinese Journal of Radiation Oncology, 2017, 26(6): 682-686.
[1] Asai A,Kawamoto K.Radiation-induced brain injury[J].Brain Ner,2008,60(2):123. [2] 刘强.放射性脑损伤研究现状[J].国外医学:放射医学核医学分册,2004,28(4):178-181.DOI:10.3760/cma.j.issn.1673-4114.2004.04.010. Liu Q.The study status of radiation induced braininjury[J].Fore Med Sci:Sect Radiat Med Nucl Med,2004,284):178-181.DOI:10.3760/cma.j.issn.1673-4114.2004.04.010. [3] Shi S,Klotz U.Clinical use and pharmacological properties of selective COX-2 inhibitors[J].Eur J Clin Pharmacol,2008,64(3):233-252.DOI:10.1007/s00228-007-0400-7. [4] 熊耀祖,涂彧,徐晓婷,等.大鼠放射性脑损伤模型中Cox-2 mRNA及蛋白的动态变化研究[J].中华放射医学与防护杂志,2016,36(1):24-27.DOI:10.3760/cma.j.issn.0254-5098.2016.01.004. Xiong YZ,Tu Y,Xu XT,et al. The dynamic changes of Cox-2 mRNA and protein expressions in the radiation injured rat brains[J].Chin J Radiol Med Protect,2016,36(1):24-27.DOI:10.3760/cma.j.issn.0254-5098.2016.01.004. [5] 杨美玉,徐晓婷,涂彧,等.塞来昔布对大鼠急性放射性脑损伤的保护作用[J].放射研究与放射工艺学报,2012(01):32-36. Yang MY,Xu XT,Tu Y,et al. The protective effect of celecoxib on radiation-induced acute brain injury in rats[J].J Radiat Res Radiat Proc,2012(01):32-36. [6] Quarmby S,Kumar P,Kumar S.Radiation-induced normal tissue injury:role of adhesion molecules in leukocyte-endothelial cell interactions[J].Int J Cancer,1999,82(3):385-395. [7] Lucas J,Mack WJ.Effects of ionizing radiation on cerebral vasculature[J].World Neurosurg,2014,81(3-4):490-491.DOI:10.1016/j.wneu.2014.01.006. [8] Li YQ,Chen P,Haimovitz-Friedman M,et al. Endothelial apoptosis initiates acute blood-brain barrier disruption after ionizing radiation[J].Cancer Res,2003,63(18):5950-5956. [9] 盛民立,朱涵能,沈立刚,等.电离放射对内皮细胞分泌TXA2和 PGI2的影响[J].细胞生物学杂志,1990(02):89-92. Sheng ML,Zhu HN,Shen LG,et al. Effects of ionizing radiation on the secretion of TXA2 and PGI2 in endothelial cells[J].Chin J Cell Biol,1990(02):89-92. [10] Vane J,Corin RE.Prostacyclin:a vascular mediator[J].Eur J Vasc Endovasc Surg,2003,26(6):571-578.DOI:10.1016/s1078. [11] Cipollone F,Cicolini G,Bucci M.Cyclooxygenase and prostaglandin synthases in atherosclerosis:recent insights and future perspectives[J].Pharmacol Ther,2008,118(2):161-180.DOI:10.1016/j.pharmthera.2008.01.002. [12] PeaLA,Fuks Z,Kolesnick RN.et al. Radiation-induced apoptosis of endothelial cells in the murine central nervous system:protection by fibroblast growth factor and sphingomyelinase deficiency[J].Cancer Res,2000,60(2):321-327. [13] Dhanasekaran DN,Reddy EP,et al. JNK signaling in apoptosis[J].Oncogene,2008,27(48):6245-6251.DOI:10.1038/onc.2008.301. [14] Petrich BG,Molkentin JD,Wang Y,et al. Temporal activation of c-Jun N-terminal kinase in adult transgenic heart via cre-loxP-mediated DNA recombi nation[J].Faseb J,2003,17(6):749-751.DOI:10.1096/fj.02-0438fje. [15] Nieminen R,Lahti A,Jalonen U,et al. JNK inhibitor SP600125 reduces COX-2 expression by attenuating mRNA in activated murine J774 macrophages[J].Int Immunopharmacol,2006,6(6):987-996.DOI:10.1016/j.intimp.2006.01.009. [16] Colangelo V,Gordon WC,Mukherjee P,et al. Downregulation of COX-2 and JNK expression after induction of ischemic tolerance in the gerbil brain[J].Brain Res,2004,1016(2):195-200.DOI:10.1016/j.brainres.2004.05.017.
2017, Vol. 26 
