Histone deacetylase 3 inhibitor ameliorates hypoxia/ reoxygenation injury in PC12 cells by reducing oxidative
10.16098/j.issn.0529-1356.2021.05.006
- Author:
Lei LEI
1
;
Yu-Zhen YOU
1
;
Yan MEI
1
;
Xuan LU
2
;
Jing-Song YANG
3
Author Information
1. Department of Medicine, Wuhan City College
2. Department of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology
3. Department of Oncology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology
- Publication Type:Journal Article
- Keywords:
Flow cytometry;
Histone deacetylase inhibitor;
Hypoxia/reoxygenation;
Oxidative stress response
- From:
Acta Anatomica Sinica
2021;52(5):706-711
- CountryChina
- Language:Chinese
-
Abstract:
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.
