皮质酮诱导的PC12细胞梯度应激损伤模型的构建及评价*

doi:10.12047/j.cjap.6286.2022.036

中国应用生理学杂志 ›› 2022, Vol. 38 ›› Issue (3): 284-288.doi: 10.12047/j.cjap.6286.2022.036

• 技术方法 • 上一篇

皮质酮诱导的PC12细胞梯度应激损伤模型的构建及评价^*

李明哲¹, 徐龙飞^1,2, 陈照立¹, 王新兴¹, 蒲玲玲¹, 刘伟丽¹, 王天辉^1△

1.军事科学院军事医学研究院环境医学与作业医学研究所, 天津 300050;
2.天津体育学院天津市运动生理与运动医学重点实验室, 天津 301617

收稿日期:2022-02-24 修回日期:2022-04-25 出版日期:2022-05-28 发布日期:2022-09-05
通讯作者: ^△Tel: 13602054718; E-mail: wydny668@163.com
基金资助:
^*十四五计划重点项目(BWS21J001);后勤科研重点项目(BWS17J025)

Construction and evaluation of a gradient stress model of PC12 cells induced by corticosterone

LI Ming-zhe ¹, XU Long-fei ^1,2, CHEN Zhao-li ¹, WANG Xin-xing ¹, PU Ling-ling¹, LIU Wei-li ¹, WANG Tian-hui ^1△

1. Tianjin Institute of Environmental and Operational Medicine, Tianjin 300050;
2. Tianjin University of Sports, Tianjin 301617, China

Received:2022-02-24 Revised:2022-04-25 Online:2022-05-28 Published:2022-09-05

摘要/Abstract

摘要： 目的: 建立皮质酮诱导的PC12细胞梯度应激损伤模型,为细胞应激水平的评估和细胞应激损伤调控研究提供实验基础和对象。方法: 通过检测不同浓度皮质酮(0~1 000 μmol/L)在经过不同干预时间(8~48 h)后PC12细胞活力,观察皮质酮对细胞活力的影响,筛选最佳干预条件的细胞模型。分光光度法和微量法检测细胞模型的关键应激指标(MDA、SOD、NADH、LDH),对模型进行评价。结果: 当皮质酮浓度在200 μmol/L以下且干预时间为12 h时,细胞活力在半数失活率以下,可减少各组由于细胞活力下降而产生的混杂因素。与空白对照组比较,皮质酮浓度依赖性地升高模型组的MDA、NADH和LDH水平,降低SOD水平(P＜0.01),符合梯度应激模型的构建要求。结论: 成功建立了PC12细胞梯度应激损伤模型,在干预时间为12 h的情况下,干预浓度为0 μmol/L、25 μmol/L、50 μmol/L、100 μmol/L、150 μmol/L、200 μmol/L,使得细胞模型应激损伤程度梯度增加,可作为开展细胞应激损伤评估及调控实验的基础和对象。

关键词: 皮质酮, 应激, PC12细胞, 细胞模型, 细胞培养

Abstract: Objective: A gradient stress model of PC12 cells induced by corticosterone was established to provide a basis for the evaluation and regulation of cell stress. Methods: The effect of corticosterone on cell viability was observed by measuring PC12 cell viability at different concentrations of corticosterone (0~1 000 μmol/L) after different intervention times (8~48 h) to screen the cell models for optimal intervention conditions. Key stress indicators (MDA, SOD, NADH, LDH) were measured spectrophotometrically and microscopically to evaluate the models. Results: When the concentration of corticosterone was below 200 μmol/L and the intervention time was 12 h, the cell viability was below half inactivation rate, which could reduce the confounding factors due to the decrease of cell viability in each group. Compared with the blank control group, corticosterone increased the levels of MDA, NADH and LDH,and decreased the levels of SOD in the model group in a concentration-dependent manner (P＜0.01), which was consistent with the construction of the gradient stress model. Conclusion: A gradient stress injury model of PC12 cells was successfully established, with intervention concentrations of 0 μmol/L, 25 μmol/L, 50 μmol/L, 100 μmol/L, 150 μmol/L and 200 μmol/L corticosterone at an intervention time of 12 h. The degree of stress injury of the cell model was increased gradually, which could be used as a basis and object for conducting cell stress injury assessment and regulation experiments.

Key words: corticosterone, stress, PC12 cells, cell model, cell culture

中图分类号:

Q-331

李明哲, 徐龙飞, 陈照立, 王新兴, 蒲玲玲, 刘伟丽, 王天辉. 皮质酮诱导的PC12细胞梯度应激损伤模型的构建及评价^*[J]. 中国应用生理学杂志, 2022, 38(3): 284-288.

LI Ming-zhe , XU Long-fei , CHEN Zhao-li , WANG Xin-xing , PU Ling-ling, LIU Wei-li , WANG Tian-hui. Construction and evaluation of a gradient stress model of PC12 cells induced by corticosterone[J]. CJAP, 2022, 38(3): 284-288.

参考文献

[1] Zeman MK, Cimprich KA. Causes and consequences of replication stress[J]. Nat Cell Biol, 2014, 16(1): 2-9.
[2] Hashimoto Y, Tanaka H. Mre11 exonuclease activity promotes irreversible mitotic progression under replication stress[J]. Life Sci Alliance, 2022, 5(6): 24-29.
[3] de Punder K, Heim C, Wadhwa PD, et al. Stress and immunosenescence: The role of telomerase[J]. Psychoneuroendocrinology, 2019, 101: 87-100.
[4] Barron H, Hafizi S, Andreazza AC, et al. Neuroinflammation and oxidative stress in psychosis and psychosis risk[J]. Int J Mol Sci, 2017, 18(3): 651-653.
[5] Barron H, Hafizi S, Mizrahi R. Towards an integrated view of early molecular changes underlying vulnerability to social stress in psychosis[J]. Mod Trends Pharmacopsychiatry, 2017, 31(1): 96-106.
[6] de Quervain D, Schwabe L, Roozendaal B. Stress, glucocorticoids and memory: implications for treating fear-related disorders[J]. Nat Rev Neurosci, 2017, 18(1): 7-19.
[7] Vitousek MN, Taff CC, Ryan TA, et al. Stress resilience and the dynamic regulation of glucocorticoids[J]. Integr Comp Biol, 2019, 59(2): 251-263.
[8] Kinlein SA, Phillips DJ, Keller CR, et al. Role of corticosterone in altered neurobehavioral responses to acute stress in a model of compromised hypothalamic-pituitary-adrenal axis function[J]. Psychoneuroendocrinology, 2019, 102: 248-255.
[9] Joëls M, Karst H, Sarabdjitsingh RA. The stressed brain of humans and rodents[J]. Acta Physiol (Oxf), 2018, 223(2): e13066.
[10] Nishi M. Effects of early-life stress on the brain and behaviors: Implications of early maternal separation in rodents[J]. Int J Mol Sci, 2020, 21(19): 246-252.
[11] Wiatrak B, Kubis-Kubiak A, Piwowar A, et al. PC12 cell line: Cell types, coating of culture vessels, differentiation and other culture conditions[J]. Cells, 2020, 9(4): 958-961.
[12] Tsikas D. Assessment of lipid peroxidation by measuring malondialdehyde (MDA) and relatives in biological samples: Analytical and biological challenges[J]. Anal Biochem, 2017, 524: 13-30.
[13] Attaluri S, Arora M, Madhu LN, et al. Oral nano-curcumin in a model of chronic gulf war illness alleviates brain dysfunction with modulation of oxidative stress, mitochondrial function, neuroinflammation, neurogenesis, and gene expression[J]. Aging Dis, 2022, 13(2): 583-613.
[14] Mansuroĝlu B, Derman S, Yaba A, et al. Protective effect of chemically modified SOD on lipid peroxidation and antioxidant status in diabetic rats[J]. Int J Biol Macromol, 2015, 72(6): 79-87.
[15] Li J, Zhang Y, Zhang J, et al. Oxidative stress and its related factors in latent autoimmune diabetes in adults[J]. Biomed Res Int, 2021, 2021: 5676363.
[16] Oldham WM, Clish CB, Yang Y, et al. Hypoxia-mediated increases in l-2-hydroxyglutarate coordinate the metabolic response to reductive stress[J]. Cell Metab, 2015, 22(2): 291-303.
[17] Li M, Liu C, Zhang W, et al. An NADH-selective and sensitive fluorescence probe to evaluate living cell hypoxic stress[J]. J Mater Chem B, 2021, 9(46): 9547-9552.
[18] Handy DE, Loscalzo J. Responses to reductive stress in the cardiovascular system[J]. Free Radic Biol Med, 2017, 109: 114-124.
[19] Bradshaw PC. Cytoplasmic and mitochondrial NADPH-coupled redox systems in the regulation of aging[J]. Nutrients, 2019, 11(3): 22-26.
[20] Sarsour EH, Kumar MG, Chaudhuri L, et al. Redox control of the cell cycle in health and disease[J]. Antioxid Redox Signal, 2009, 11(12): 2985-3011.
[21] Clanton TL. Hypoxia-induced reactive oxygen species formation in skeletal muscle[J]. J Appl Physiol, (1985) 2007, 102(6): 2379-2388.
[22] Jurisic V, Radenkovic S, Konjevic G. The actual role of LDH as tumor marker, biochemical and clinical aspects[J]. Adv Exp Med Biol, 2015, 867(3): 115-124.
[23] Wang C, Wang XR, Song DD, et al. The establishment of rat model in myocardial ischemia with psychological stress[J]. Ann Transl Med, 2020, 8(6): 322.

皮质酮诱导的PC12细胞梯度应激损伤模型的构建及评价^*

Construction and evaluation of a gradient stress model of PC12 cells induced by corticosterone

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