组蛋白去乙酰化酶3抑制剂通过减轻氧化应激降低缺氧/复氧引起的PC12细胞凋亡

doi:10.16098/j.issn.0529-1356.2021.05.006

解剖学报 ›› 2021, Vol. 52 ›› Issue (5): 706-711.doi: 10.16098/j.issn.0529-1356.2021.05.006

组蛋白去乙酰化酶3抑制剂通过减轻氧化应激降低缺氧/复氧引起的PC12细胞凋亡

雷蕾^1* 陆铉²杨劲松³尤玉珍¹ 梅燕¹

1.武汉城市学院医学部，武汉 430083; 2.华中科技大学同济医学院附属协和医院血液内科，武汉 430022; 3.华中科技大学同济医学院附属协和医院肿瘤科，武汉 430022

收稿日期:2020-04-09 修回日期:2020-05-22 出版日期:2021-10-06 发布日期:2021-10-06
通讯作者: 雷蕾 E-mail:lei6423@163.com

Histone deacetylase 3 inhibitor ameliorates hypoxia/reoxygenation injury in PC12 cells by reducing oxidative#br#

LEI Lei^1* LU Xuan²YANG Jing-song³ YOU Yu-zhen¹ MEI Yan¹

1.Department of Medicine, Wuhan City College, Wuhan 430083, China; 2.Department of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China; 3.Department of Oncology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China

Received:2020-04-09 Revised:2020-05-22 Online:2021-10-06 Published:2021-10-06
Contact: LEI Lei E-mail:lei6423@163.com

摘要/Abstract

摘要：

目的探讨组蛋白去乙酰化酶3（HDAC3）抑制剂（HDAC3I）RGFP966在PC12细胞缺氧/复氧（H/R)损伤中的作用。方法采用PC12细胞缺氧4 h复氧24 h培养建立H/R细胞损伤模型。H/R+抑制剂组采用RGFP966预处理1 h后进行H/R处理。实验分为3组，对照组、H/R组和H/R+抑制剂组，每组重复3次。采用MTT法测定细胞活性，比色法检测细胞乳酸脱氢酶(LDH)，流式细胞术分别检测细胞凋亡率和胞内活性氧簇（ROS），黄嘌呤氧化酶法测定超氧化物歧化酶（SOD）活性，硫代巴比妥酸法测定丙二醛（MDA）含量，Western blotting法检测Bcl-2促凋亡基因(Bax)、B淋巴细胞瘤-2基因（Bcl-2）、剪切型Caspase-3（cleaved-Caspase-3）和HDAC3蛋白表达。结果与对照组相比，H/R组与H/R+抑制剂组细胞活力均显著降低（P<0.05），细胞LDH（P<0.05）和凋亡率（P<0.05）均显著升高。H/R+抑制剂组与H/R组相比，H/R+抑制剂组细胞活力显著高于H/R组（P<0.05），而H/R+抑制剂组细胞LDH（P<0.05）和凋亡率显著低于H/R组（P<0.05）。此外，与对照组相比，H/R组与H/R+抑制剂组ROS和MDA（P<0.05）均显著增加，SOD显著降低（P<0.05）；H/R+抑制剂组与H/R组相比，H/R+抑制剂组ROS和MDA（P<0.05）显著低于H/R组，而SOD水平高于H/R组（P<0.05）；Western blotting结果表明，与对照组相比，H/R组与H/R+抑制剂组Bax、cleaved-Caspase-3均显著升高，Bcl-2显著降低（P<0.05），H/R+抑制剂组Bax、cleaved-Caspase-3均显著低于H/R组，Bcl-2显著高于H/R组（P<0.05）；与对照组相比，H/R组HDAC3蛋白表达显著升高（P<0.05），而H/R+抑制剂组HDAC3蛋白显著降低（P<0.05）。
结论 HDAC3I通过减轻氧化应激降低缺氧/复氧引起的PC12细胞凋亡。

关键词: 组蛋白去乙酰化酶抑制剂, 氧化应激反应, 缺氧/复氧, 流式细胞术

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 PC12 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 PC12 cell apoptosis induced by hypoxia/reoxygenation by reducing oxidative stress.

Key words: Histone deacetylase inhibitor, Oxidative stress response, Hypoxia/reoxygenation, Flow cytometry

中图分类号:

R363

雷蕾陆铉杨劲松尤玉珍梅燕. 组蛋白去乙酰化酶3抑制剂通过减轻氧化应激降低缺氧/复氧引起的PC12细胞凋亡[J]. 解剖学报, 2021, 52(5): 706-711.

LEI Lei LU Xuan YANG Jing-song YOU Yu-zhen MEI Yan. Histone deacetylase 3 inhibitor ameliorates hypoxia/reoxygenation injury in PC12 cells by reducing oxidative#br#[J]. Acta Anatomica Sinica, 2021, 52(5): 706-711.

参考文献

［1］ Wu YJ, Chen ChH. Progress of adjuvant treatment with tissue plasminogen activator beyond time window in ischemic stroke［J］. Acta Anatomica Sinica, 2019,50(6):850-856.(in Chinese)
吴冶君,陈春花.缺血性脑卒中治疗时间窗后辅助溶栓治疗的进展［J］.解剖学报,2019,50(6):850-856.
［2］ Cao DF, Yang N. Structure and catalytic mechanisms of histone deacetylases［J］. Progress in Biochemistry and Biophysics,2015, 42(11):978-993. (in Chinese)
曹端方,杨娜.组蛋白去乙酰化酶的结构及应用［J］.生物化学与生物物理进展,2015,42(11):978-993.
［3］ Yang X, Wu Q, Zhang L, et al. Inhibition of histone deacetylase 3 (HDAC3) mediates ischemic preconditioning and protects cortical neurons against ischemia in rats ［J］. Front Mol Neurosci, 2016,9:131.
［4］ Demyanenko S, Neginskaya M, Berezhnaya E. Expression of class Ⅰ histone Deacetylases in ipsilateral and contralateral hemispheres after the focal photothrombotic infarction in the mouse brain ［J］.Transl Stroke Res, 2018, 9(5):471-483.
［5］ Jiang R, Lü KN, Pan XF, et al. Current status and challenges of epigenetic drug research and development［J］. Biotechnology Bulletin, 2019,35(8):213-225. (in Chinese)
江芮,吕柯孬,潘学峰,等.表观遗传药物研发的现状与挑战［J］.生物技术通报,2019,35(8):213-225.
［6］ Dong ZhJ, Han Ch, Liu JJ, et al. Histone deacetylase inhibitors:research advances［J］. Journal of International Pharmaceutical Research, 2017,44(12):1098-1106, 1124. (in Chinese)
董照记,韩诚,刘晶晶,等.组蛋白去乙酰化酶抑制剂的研究进展［J］.国际药学研究杂志,2017,44(12):1098-1106,1124.
［7］ Li L, Jin J, Yang XJ. Histone deacetylase 3 governs perinatal cerebral development via neural stem and progenitor cells ［J］. iScience, 2019,20:148-167.
［8］ Zhan HY, Tao H, Xu ShS, et al. Role of HDAC3 in H9C2 cell ischemia-reperfusioninjury and intervention mechanism ［J］. Acta Universitatis Medicinalis Anhui, 2017,52(5):674-677. (in Chinese)
占红英,陶辉,徐盛松,等. HDAC3在H9C2细胞缺血再灌注损伤中的表达及干预机制［J］.安徽医科大学学报,2017,52(5):674-677.
［9］ Bardai FH, D’Mello SR. Selective toxicity by HDAC3 in neurons: regulation by Akt and GSK3beta ［J］. J Neurosci, 2011,31(5):1746-1751.
［10］ Zhao QCh, Xu Y. Research progress of histone deacetylase 3 in nervous system diseases ［J］. Journal of International Neurology and Neurosurgery, 2015, 42(4):384-388. (in Chinese)
赵秋宸,徐运.组蛋白去乙酰化酶3在神经系统疾病中的研究进展［J］.国际神经病学神经外科学杂志,2015,42(4):384-388.
［11］ Fan WX. Research progress on the mechanism of ischemic stroke ［J］. Journal of China Pharmaceutical University, 2018, 49(6):751-759. (in Chinese)
樊文香.缺血性脑卒中的机制研究进展［J］.中国药科大学学报,2018,49(6):751-759.
［12］ Chen H, Wu JJ, Xue Q, et al. Progress in research on the mitochondrial mechanism of ischemica stroke and effective components of TCM［J］. Chinese Journal of New Drugs, 2016, 25(11):1236-1240. (in Chinese)
陈红,吴俊杰,薛强,等.以线粒体为靶点抗缺血性脑卒中中药有效成分研究进展［J］.中国新药杂志,2016,25(11):1236-1240.

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组蛋白去乙酰化酶3抑制剂通过减轻氧化应激降低缺氧/复氧引起的PC12细胞凋亡

Histone deacetylase 3 inhibitor ameliorates hypoxia/reoxygenation injury in PC12 cells by reducing oxidative#br#

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