Protective effect of thymosin ß4 on oxygen-glucose deprivation/reoxygenation injury in rat cortical neurons / 中国脑血管病杂志

ABSTRACT

Objective To investigate the protective effect and mechanisms of thymosin beta 4 (Tß4) on oxygen-glucose deprivation/reoxygenation (O G D / R) injury in rat cortical neurons. Methods Primary cultured cortical neurons were isolated and identified; Ischemia-reperfusion injury model of rat cortical neurons was established (oxygen glucose deprivation 6 h, reoxygenation 12 h) by OGD / R. The rats were divided into the control group,the model group and the treatment group (addition of thymosinß4 2 h before modeling); The cell counting Kit-8 (CCK8) was used to determine the optimal concentration of Tß4. Flow cytometry and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) were used to detect apoptosis. Western blotting was used to detect the expression of 78 000 glucose-regulated protein 78 (GRP 78), CCAAT/enhancer binding protein-homologous protein (CHOP), B-cell lymphoma-2 (Bel-2) and Bax. Expression levels were compared among groups. One-way analysis of variance was used to compare the normally distributed measurement data among three groups with the SPSS19. 0 software. Results Cortical neurons were separated correctly. The optimal concentration of Tß4 was 10 (ig/L. Comparing the model group with the control group, the survival rate of cortical neurons decreased significantly (P = 0. 002), the apoptotic rate increased significantly (P < 0. 01), the expression of GRP78,CHOP and Bax was up-regulated significantly (P values were 0. 034,0 and 0. 045 .respectively),and the expression of Bcl-2 was reduced significantly (P = 0. 006). Comparing the treatment group(10 p.g/L exogenous Tß4) with the model group, cell viability increased significantly (P = 0. 008), the apoptotic rate decreased significantly (P = 0. 002),the expression of GRP78,CHOP and Bax decreased significantly (P values were 0. 032,0. 027 and 0. 019, respectively),and the expression of Bcl-2 increased significantly (P = 0.028) .The differences were statistically significant. Conclusions Tß4 inhibits OGD/R-induced endoplasmic reticulum stress-dependent apoptosis. These results provide a theoretical basis for the application of Tß4 in the treatment of cerebral ischemia-reperfusion injury.