Relationship between TDAG8 and endogenous neuron-protective mechanism against oxygen-glucose deprivation and restoration-induced apoptosis in rat neurons
10.3760∕cma.j.issn.0254-1416.2016.09.011
- VernacularTitle:TDAG8与氧糖缺失-复糖复氧诱发大鼠神经元凋亡时内源性保护机制的关系
- Author:
Xiaodong MA
;
Donghua SHAO
;
Lihua HANG
;
Weiwei SHU
;
Xiulan HU
;
Hong LUO
- Keywords:
Receptors,G-protein-coupled;
Apoptosis;
Neurons;
Reperfusion injury
- From:
Chinese Journal of Anesthesiology
2016;36(9):1080-1084
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the relationship between T?cell death?associated gene 8 ( TD?AG8) and endogenous neuron?protective mechanism against oxygen?glucose deprivation and restoration ( OGD∕R)?induced apoptosis in rat neurons. Methods The primary cortical neurons obtained from fetal rats were seeded in 6?well plates at a density of 1×105 cells∕ml and divided into 5 groups using a random number table: control group ( group C, n=24 ) , group OGD∕R ( n=48 ) , TDAG8 agonist BTB09089 group (group BTB, n=24), TDAG8?siRNA group ( group siRNA, n=24), and blank vehicle group ( group V, n=24) . The medium was replaced with glucose?and serum?free Locke′s buffer, and the neu?rons were exposed to 95% N2?5% CO2 in an air?tight incubator at 37℃ for 60 min followed by routine cul?ture to establish the model of OGD∕R. In BTB, siRNA and V groups, 20 μmol∕L TDAG8 agonist
BTB09089, 200 pmol∕L TDAG8?siRNA, and 6 μl∕200 μl transfection reagent were added, respectively, at 24 h before oxygen?glucose restoration. At 6 h of oxygen?glucose restoration, the neuronal viability and a?mount of lactic dehydrogenase ( LDH) released were measured, and the expression of TDAG8 and caspase?3 mRNA in neurons was detected by fluorescent quantitative real?time polymerase chain reaction. In group OGD∕R, the expression of TDAG8 and caspase?3 was measured by Western blot at 0, 3, 6, 12 and 24 h of oxygen?glucose restoration. In C, OGD∕R, BTB, siRNA and V groups, the expression of TDAG8, caspase?3 and p?Akt was detected at 6 h of oxygen?glucose restoration. Results In group OGD∕R, the ex?pression of TDAG8 was gradually up?regulated after oxygen?glucose restoration, and the expression of caspase?3 peaked at 6 h of oxygen?glucose restoration. Compared with group C, the neuronal viability was significantly decreased, the amount of LDH released was significantly increased, and the expression of TD?AG8 and caspase?3 protein and mRNA and p?Akt was significantly up?regulated in OGD∕R, V and siRNA groups ( P<0?05) . Compared with group OGD∕R, the expression of TDAG8 protein and mRNA and p?Akt was significantly up?regulated, the expression of caspase?3 protein and mRNA was significantly down?regu?lated, the neuronal viability was significantly increased, and the amount of LDH released was significantly decreased in group BTB, the expression of TDAG8 protein and mRNA and p?Akt was significantly down?regulated, the expression of caspase?3 protein and mRNA was significantly up?regulated, the neuronal via?bility was significantly decreased, and the amount of LDH released was significantly increased in group siR?NA ( P<0?05) , and no significant change was found in the parameters mentioned above in group V ( P>0?05) . Conclusion TDAG8 is partially involved in the endogenous neuron?protective mechanism against OGD∕R?induced apoptosis in rat neurons, which may be related to activation of Akt signaling pathway.
