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; 陆铉; 杨劲松; 尤玉珍; 梅燕

doi:10.16098/j.issn.0529-1356.2021.05.006

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

神经生物学

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

作者信息 +

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

Author information +

文章历史 +

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。

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.

导出引用

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 https://doi.org/10.16098/j.issn.0529-1356.2021.05.006

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 https://doi.org/10.16098/j.issn.0529-1356.2021.05.006

中图分类号： R363

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