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ARTICLE

Hyperbaric oxygen protects against PC12 and H9C2 cell damage caused by oxygen–glucose deprivation/reperfusion via the inhibition of cell apoptosis and autophagy

JIANRONG YANG^1,#, WAN CHEN^2,#, XING ZHOU^3,#, YAOXUAN LI^4,#, ZHIHUANG NONG⁵, LIYUAN ZHOU⁴, XUAN WEI⁴, XIAORONG PAN⁶, CHUNXIA CHEN⁷, WENSHENG LU³

1 Hepatobiliary Surgery, Guangxi Academy of Medical Sciences, The People’s Hospital of Guangxi Zhuang Autonomous Region, Nanning, 530021, China
2 Department of Emergency, The People’s Hospital of Guangxi Zhuang Autonomous Region, Nanning, 530021, China
3 Department of Endocrinology, The People’s Hospital of Guangxi Zhuang Autonomous Region, Nanning, 530021, China
4 Department of Neurology, The People’s Hospital of Guangxi Zhuang Autonomous Region, Nanning, 530021, China
5 Department of Pharmacy, The People’s Hospital of Guangxi Zhuang Autonomous Region, Nanning, 530021, China
6 Department of Hyperbaric Oxygen, The People’s Hospital of Guangxi Zhuang Autonomous Region, Nanning, 530021, China
7 Department of Research Center of Medical Sciences, The People’s Hospital of Guangxi Zhuang Autonomous Region, Nanning, 530021, China

* Corresponding Authors:CHUNXIA CHEN. Email: ; WENSHENG LU. Email:
# These authors contributed equally to this work

(This article belongs to the Special Issue: Cellular Biomechanics in Health and Diseases)

BIOCELL 2022, 46(1), 137-148. https://doi.org/10.32604/biocell.2022.016807

Received 28 March 2021; Accepted 14 May 2021; Issue published 28 September 2021

Abstract

In this study, we investigated the protective effect of hyperbaric oxygen (HBO) on PC12 and H9C2 cell damage caused by oxygen-glucose deprivation/reperfusion and its possible mechanism. PC12 and H9C2 cell oxygen-glucose deprivation/reperfusion model were established. Cells were divided into a control group, model group, hyperbaric air (HBA) group and HBO group. The cell viability was detected by the CCK8 assay. Hoechst 33342 and PI staining assays and mitochondrial membrane potential (MMP) assays were used to detect cell apoptosis. The ultrastructure of cells, including autophagosomes, lysosomes, and apoptosis, were examined using a transmission electron microscope. The expression of autophagy-related proteins was detected by cellular immunofluorescence and immunocytochemistry. Our results showed that HBO can significantly improve the vitality of damaged PC12 and H9C2 cells caused by oxygen–glucose deprivation/reperfusion. HBO can significantly inhibit apoptosis of PC12 and H9C2 cells caused by oxygen-glucose deprivation/reperfusion. Importantly, we found that the protective mechanism of PC12 and H9C2 cell damage caused by oxygen-glucose deprivation/reperfusion may be related to the inhibition of the autophagy pathway. In this study, the results of cellular immunofluorescence and immunocytochemistry experiments showed that the 4E-BP1, p-AKt and mTOR levels of PC12 and H9C2 cells in the model group decreased, while the levels of LC3B, Atg5 and p53 increased. However, after HBO treatment, these autophagy-related indexes were reversed. In addition, observation of the cell ultrastructure with transmission electron microscopy found that in the model group, a significant increase in the number of autophagic vesicles was observed. In the HBO group, a decrease in autophagic vesicles was observed. The study demonstrated that hyperbaric oxygen protects against PC12 and H9C2 cell damage caused by oxygen-glucose deprivation/reperfusion via the inhibition of cell apoptosis and autophagy.

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