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J Zhejiang Univ (Med Sci)  2021, Vol. 50 Issue (5): 614-620    DOI: 10.3724/zdxbyxb-2021-0343
Establishment of a hypobaric hypoxia-induced cell injury model in PC12 cells
ZHANG Dongmei1,CAO Qilu1,JING Linlin1,ZHAO Xiuhua2,*(),MA Huiping1,*()
1. Key Laboratory of the Plateau Medicine, the 940th Hospital of Joint Logistics Support Force of Chinese People’s Liberation Army, Lanzhou 730050, China;
2. Second Department of Infectious Disease Control and Prevention, Center for Disease Control and Prevention, Western War Zone, Chengdu 610000, China
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Objective: To construct a hypobaric hypoxia-induced cell injury model. Methods:Rat pheochromocytoma PC12 cells were randomly divided into control group, normobaric hypoxia group and hypobaric hypoxia group. The cells in control group were cultured at normal condition, while cells in other two groups were cultured in normobaric hypoxia and hypobaric hypoxia conditions, respectively. CCK-8 method was used to detect cell viability to determine the optimal modeling conditions like the oxygen concentration, atmospheric pressure and low-pressure hypoxia time. The contents of lactate dehydrogenase (LDH), superoxide dismutase (SOD) and malondialdehyde (MDA) were detected by microplate method. The apoptosis ratio and cell cycle were analyzed by flow cytometry. Results: The hypobaric hypoxia-induced cell injury model can be established by culturing for 24?h at 1% oxygen concentration and 41?kPa atmospheric pressure. Compared with the control group and normobaric hypoxia group, the activity of LDH and the content of MDA in hypobaric hypoxia group were significantly increased, the activity of SOD was decreased, the percentage of apoptosis was increased (all P<0.05), and the cell cycle was arrested in G0/G1 phase.Conclusion: A stable and reliable cell injury model induced by hypobaric hypoxia has been established with PC12 cells, which provides a suitable cell model for the in vitro experimental study on nerve injury induced by hypoxia at high altitude.

Key wordsPC12 cells      Hypobaric hypoxia      Lactate dehydrogenase      Superoxide dismutase      Malondialdehyde      Apoptosis      Cell cycle     
Received: 20 February 2021      Published: 29 December 2021
CLC:  R966  
Corresponding Authors: ZHAO Xiuhua,MA Huiping     E-mail:
Cite this article:

ZHANG Dongmei,CAO Qilu,JING Linlin,ZHAO Xiuhua,MA Huiping. Establishment of a hypobaric hypoxia-induced cell injury model in PC12 cells. J Zhejiang Univ (Med Sci), 2021, 50(5): 614-620.

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关键词: PC12细胞,  低压性缺氧,  乳酸脱氢酶,  超氧化物歧化酶,  丙二醛,  细胞凋亡,  细胞周期 
Figure 1 Effects of different hypoxia concentrations on the activity of PC12 cells
Figure 2 Effects of atmospheric pressure on PC12 cell activity under 1% oxygen concentration
Figure 3 Effects of different culture time on PC12 cell activity under hypobaric hypoxia condition
Figure 4 Morphology of PC12 cells cultured under different conditions for 24 h








22.55 ±2.11





38.15 ±1.99*





43.15 ±3.25**#



Table 1 Lactate dehydrogenase activity, malondialdehyde content and superoxide dismutase activity in each group
Figure 5 Results of apoptosis detected by flow cytometry
Figure 6 Cell cycles of PC12 cells in each group
[1]   ZHANGP, CHENJ S, LIQ Y, et al.Neuroprotectants attenuate hypobaric hypoxia-induced brain injuries in cynomolgus monkeys[J]Zool Res, 2021, 42( 1): 3-19.
doi: 10.24272/j.issn.2095-8137.2020.384
[2]   SHARMAP, TULSAWANIR. Ganoderma lucidum aqueous extract prevents hypobaric hypoxia induced memory deficit by modulating neurotransmission, neuroplasticity and maintaining redox homeostasis[J]Sci Rep, 2020, 10( 1): 8944.
doi: 10.1038/s41598-020-65812-5
[3]   陈建, 黄缄, 高钰琪, 等. 预缺氧对PC12细胞缺氧后细胞内游离Ca2+的影响[J]. 西北国防医学杂志, 2008, 29(4): 250-253
CHEN Jian, HUANG Jian, GAO Yuqi, et al. Effect of hypoxia preconditioning on intracellular Ca2+ of PC12 cells[J]. Medical Journal of National Defending Forces in Northwest China, 2008, 29(4): 250-253. (in Chinese)
[4]   闫向丽, 王利胜, 王圣鑫. HT22海马神经元细胞氧糖剥夺再灌注模型的建立[J]. 广州中医药大学学报, 2020, 37(181): 161-165
YAN Xiangli, WANG Lisheng, WANG Shengxin. Establishment of HT22 cell oxygen-glucose deprivation reperfusion model[J]. Journal of Guangdong Pharmaceutical University, 2020, 37(181): 161-165. (in Chinese)
[5]   刘菲, 张昊, 刘博, 等. 氯化钴诱导的N2a细胞缺氧损伤模型的机制研究[J]. 广东药科大学学报, 2020, 36(2): 249-253
LIU Fei, ZHANG Hao, LIU Bo, et al. Mechanism of cobalt chloride induced hypoxia on N2a cells[J]. Journal of Guangzhou University of Traditional Chinese Medicine, 2020, 36(2): 249-253. (in Chinese)
[6]   李海丽, 卢永昌, 钱帅, 等. 苞叶雪莲粗提物对EA. hy926细胞缺氧损伤的保护作用[J]. 湖北农业科学, 2016, 55(1): 167-169
LI Haili, LU Yongchang, QIAN Shuai, et al. Saussurea obvallata crude protective effects of hypoxia on EA. hy926 injury[J]. Hubei Agricultural Sciences, 2016, 55(1): 167-169. (in Chinese)
[7]   HEILJ, SCHULTZED, SCHEMMERP, et al.N-acetylcysteine protects hepatocytes from hypoxia-related cell injury[J]Clin Exp Hepatol, 2018, 4( 4): 260-266.
doi: 10.5114/ceh.2018.80128
[8]   冯岚, 张二龙, 高钰琪, 等. 7种不同组织来源细胞缺氧损伤效应评价[J]. 第三军医大学学报, 2019, 41(6): 570-574
FENG Lan, ZHANG Erlong, GAO Yuqi, et al. Evaluation of hypoxic injuries in 7 cell lines derived from different human tissues[J]. Journal of Third Military Medical University, 2019, 41(6): 570-574. (in Chinese)
[9]   牛其芳, 李德龙, 杨杨, 等. 人血管内皮细胞缺氧复氧损伤细胞模型的建立[J]. 中国口腔颌面外科杂志, 2019, 17(4): 295-299
NIU Qifang, LI Delong, YANG Yang, et al. Establishment of human vascular endothelial hypoxia/reoxygeneration injury cell model[J]. China Journal of Oral and Maxillofacial Surgery, 2019, 17(4): 295-299. (in Chinese)
[10]   刘阳, 安美文, 芦莹, 等. 压力下角质形成细胞与成纤维细胞共培养对细胞增殖及胶原合成的影响[J]. 医用生物力学, 2015, 30(2): 148-153
LIU Yang, AN Meiwen, LU Ying, et al. Effects of keratinocytes/fibroblasts co-culture under pressure on cell proliferation and collagen synthesis[J]. Journal of Medical Biomechanics, 2015, 30(2): 148-153. (in Chinese)
[11]   黄正, 刘益杰, 褚立希, 等. 周期性压力通过凋亡基因调节兔软骨细胞增殖和凋亡的实验研究[J]. 国际骨科学杂志, 2014, 35(3): 205-208
HUANG Zheng, LIU Yijie, CHU Lixi, et al. Periodic pressure regulates the proliferation and apoptosis of rabbit chondrocytes genes[J]. International Journal of Orthopaedics, 2014, 35(3): 205-208. (in Chinese)
[12]   COBELENSP M, VAN PUTTEB P, KAVELAARSA, et al.Inflammatory consequences of lung ischemia-reperfusion injury and low-pressure ventilation[J]J Surg Res, 2009, 153( 2): 295-301.
doi: 10.1016/j.jss.2008.04.022
[13]   TSENGV, NIK, ALLAWZIA, et al.Extracellular superoxide dismutase regulates early vascular hyaluronan remodeling in hypoxic pulmonary hypertension[J]Sci Rep, 2020, 10( 1): 280.
doi: 10.1038/s41598-019-57147-7
[14]   ASANOA, UEDAS, KUJII, et al.Intracellular hypoxia measured by 18F-fluoromisonidazole positron emission tomography has prognostic impact in patients with estrogen receptor-positive breast cancer[J]Breast Cancer Res, 2018, 20( 1): 78.
doi: 10.1186/s13058-018-0970-6
[15]   SHIQ, YUC, ZHUD, et al.Effects of dietary Sargassum horneri on resisting hypoxia stress, which changes blood biochemistry, antioxidant status, and hepatic HSP mRNA expressions of juvenile black sea bream Acanthopagrus Schlegelii[J]J Appl Phycol, 2020, 32( 5): 3457-3466.
doi: 10.1007/s10811-020-02132-1
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