Effect of T cell death associated gene 8 on focal cerebral ischemiareperfusion injury in rats
LUO Hong1, SHAO Dong-hua1, HU Xiu-lan2, MA Xiao-dong1
(1. Department of Anesthesiology, 2.Department of Intensive Care Unit, Affiliated People’s Hospital of Jiangsu University, Zhenjiang Jiangsu 212002,China)
Abstract：Objective: To investigate the effect of activated T cell death associated gene 8(TDAG8) on focal cerebral ischemia-reperfusion injury in rats. Methods: One hundred and thirty-two healthy adult male Sprague-Dawley rats were divided into 6 groups using a random number table: control group(Group C, n=18), ischemia-reperfusion group(Group I/R, n=42), TDAG8 agonist BTB09089 group(Group BTB, n=18), blank vector group(Group V, n=18), TDAG8-siRNA negative control group(n=18) and TDAG8-siRNA group(Group siRNA, n=18). Middle cerebral artery occlusion(MCAO) was used to establish rat focal cerebral ischemia-reperfusion model. Group BTB, Group V, Group siRNA (-) and Group siRNA were injected into the lateral ventricle with 8 μL of 20 μmol/L BTB09089, 8 μL blank transfection reagent jetPEI, 4 μg siRNA negative control and 4 μg siRNA, respectively, at 72 h, 48 h and 24 h before ischemia. The Group V, Group siRNA (-) and Group siRNA contained the same amount of transfection reagent jetPEI. At 6 h after reperfusion, the brain was decapitated, the expression of TDAG8, pro-apoptotic factor cleaved caspase-3, anti-apoptotic factor Bcl-2, p-Akt and p-CREB in ischemic penumbra was detected by Western blotting. The neurological deficit score and infarct volume percentage were respectively measured at 24 h and 72 h after reperfusion. Results: Compared with Group C, at 6 h after reperfusion, the expression of TDAG8, cleaved caspase-3,p-Akt and p-CREB was up-regulated(P<0.05) and Bcl-2 expression was down-regulated(P<0.05) in Group I/R, Group BTB, Group V, Ggroup siRNA(-) and Group siRNA, and at 24 h and 72 h after reperfusion, the infarct volume percentage increased and the neurological deficit score reduced(P<0.05). Compared with Group I/R, at 6 h after reperfusion, the expression of TDAG8, Bcl-2, p-Akt and p-CREB was up-regulated(P<0.05) and cleaved caspase-3 expression was down-regulated(P<0.05) in Group BTB, and at 24 h and 72 h after reperfusion, the infarct volume percentage reduced and the neurological deficit score increased(P<0.05). Compared with Group I/R, at the 6 h after reperfusion, the expression of TDAG8, Bcl-2, p-Akt and p-CREB was down-regulated(P<0.05) and cleaved caspase-3 expression was up-regulated (P<0.05) in Group siRNA, and at 24 h and 72 h after reperfusion, the infarct volume percentage increased and the neurological deficit score reduced(P<0.05). Conclusion: Activation of TDAG8 may be involved in focal cerebral ischemia-reperfusion injury induced by transient middle cerebral artery occlusion in rats by the Akt signaling pathway.
［Key words］T-cell death associated gene 8; reperfusion injury; BTB09089; siRNA
2019, Vol. 29 
