RESUMEN
Objective To investigate the role of histone deacetylase 3 ( HDAC3 ) inhibitor (HDAC3I) in hypoxia-reoxygenation( H/R) injury of PC12 cells. Methods H/R cell injury model was established by using PC 12 cells for 4 hours hypoxia and then reoxygenation for 24 hours. HDAC3I treatment group was pretreated with RGFP966 for 1 hour and then subjected to hypoxia-reoxygenation injury. The experiment was divided into three groups: normal control group, model group and HDAC3I treatment group, and 3 repetitions for each group. Cell viability was determined using MTT. Cellulose dehydrogenase (LDH) was detected by colorimetry. Flow cytometry was used to detect apoptosis and intracellular reactive oxygen species ( ROS ), respectively. The activity of superoxide dismutase ( SOD ) was determined by xanthine oxidase method. The malondialdehyde ( MDA ) content was determined by the thiobarbituric acid method. Western blotting was used to detect the expression of Bax, Bcl-2, cleaved-Caspase-3 and HDAC3 proteins. Results Compared with the control group, the cell viability of the model group and HDAC3I treatment group decreased significantly (P<0. 05), and the cell LDH (P<0. 05) and apoptosis (P<0. 05) increased significantly. The cell viability of HDAC3I treatment group was significantly higher than that of the model group (P<0. 05), while the LDH (P<0. 05) and apoptosis of HDAC3I treatment group were lower than the model group (P<0. 05). In addition, compared with the control group, the ROS and MDA (P< 0. 05 ) of the model group and the HDAC3I treatment group increased significantly, and the SOD decreased significantly (P<0. 05). ROS and MDA in the HDAC3I treatment group (P<0. 05) were significantly lower than the model group, while the SOD level was higher than the model group (P< 0. 05). Western blotting analysis showed that compared with the control group, Bax and cleaved-Caspase-3 in the model group and HDAC3I treatment group increased significantly, and Bcl-2 decreased significantly (P<0. 05). The Bax and cleaved-Caspase-3 in the HDAC3I treatment group were significantly lower than the model group, and Bcl-2 was significantly higher than the model group (P<0. 05). Compared with the control group, the expression of HDAC3 protein in the model group increased significantly (P<0. 05), while the HDAC3 protein in the HDAC3I treatment group decreased significantly (P<0. 05). Conclusion HDAC3I reduces PC 12 cell apoptosis induced by hypoxia/reoxygenation by reducing oxidative stress.