The effects of Celecoxib on human brain microvascular endothelial cells release 6-keto-PGF1α and TXB2 and apoptosis after radiation / 中华放射肿瘤学杂志

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.