7种不同组织来源细胞缺氧损伤效应评价
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摘要
目的观察缺氧条件下7种不同类型细胞损伤程度,筛选对缺氧损伤敏感的细胞模型。方法选择CCD 841 CoN、GES-1、HUVEC、HEB、HEK-293、HBE和L02等7种不同组织来源的细胞,利用低氧工作站构建不同缺氧浓度(5%O_2和1%O_2)细胞缺氧模型,使用CCK-8和LDH方法检测细胞增殖及损伤情况。并用红景天(0.625μmol/L、2.5 mmol/L)和丁苯酞(1、2μmol/L)于缺氧开始时处理细胞,观察是否可改善缺氧细胞损伤。结果与21%O_2相比,1%O_2处理24 h时,HEB细胞增殖能力显著降低(P<0.05);与21%O_2相比,5%O_2与1%O_2处理24 h HEB、HUVEC、HBE、HEK-293、CCD 841 CoN、GES-1和L02细胞其上清中LDH酶活力均显著升高(P<0.05,P<0.01);在HEB细胞中,红景天(2.5 mmol/L)和丁苯酞(1μmol/L)处理可显著逆转低氧诱导的细胞增殖和细胞活力降低(P<0.05);红景天(2.5 mmol/L)和丁苯酞(1μmol/L)处理可显著逆转低氧诱导的LDH酶活力升高(P<0.05,P<0.01)。结论 HEB细胞对缺氧最为敏感,是较好的抗缺氧损伤药物效应研究细胞的模型。
Objective To observe hypoxic injuries in 7 cell lines derived from different human tissues to identify a cell model that is sensitive to acute hypoxic injury. Methods Seven cell lines derived from different human tissues, namely CCD 841 CoN, GES-1, HUVEC, HEB, HEK-293, HBE and L02, were exposed to different oxygen concentrations(1% and 5%) for 24 h using a hypoxia workstation. CCK-8 and LDH kits were used to assess the cell proliferation and cell injuries. The cells were then treated with rhodiola(0.625 μmol/L, 2.5 mmol/L) and 3-n-butylphthalide(1,2 μmol/L) at the start of hypoxic exposure to assess the protective effects of the drugs against hypoxic cell injuries. Results Compared with the cells cultured in normal oxygen concentration(21%), HEB cells exposed to 1% O_2 for 24 h showed significantly lowered proliferative activity(P<0.05). Hypoxic exposure to 5% or 1% O_2 for 24 h significantly increased LDH activity in the culture supernatant of HEB, HUVEC, HBE, HEK-293, CCD 841 CoN, GES-1 and L02 cells(P<0.05 or 0.01). In HEB cells cultured under hypoxic conditions, treatment with rhodiola and 3-n-butylphthalide significantly promoted the cell proliferation and improved the cell viability(P<0.05), and obviously reversed hypoxia-induced increase in LDH activity(P<0.05 or P<0.01). Conclusion Among the 7 cell lines tested in this study, HEB cells are the most sensitive to hypoxia and can serve as an ideal model for studying drug interventions of hypoxic cell injuries.
Objective To observe hypoxic injuries in 7 cell lines derived from different human tissues to identify a cell model that is sensitive to acute hypoxic injury. Methods Seven cell lines derived from different human tissues, namely CCD 841 CoN, GES-1, HUVEC, HEB, HEK-293, HBE and L02, were exposed to different oxygen concentrations(1% and 5%) for 24 h using a hypoxia workstation. CCK-8 and LDH kits were used to assess the cell proliferation and cell injuries. The cells were then treated with rhodiola(0.625 μmol/L, 2.5 mmol/L) and 3-n-butylphthalide(1,2 μmol/L) at the start of hypoxic exposure to assess the protective effects of the drugs against hypoxic cell injuries. Results Compared with the cells cultured in normal oxygen concentration(21%), HEB cells exposed to 1% O_2 for 24 h showed significantly lowered proliferative activity(P<0.05). Hypoxic exposure to 5% or 1% O_2 for 24 h significantly increased LDH activity in the culture supernatant of HEB, HUVEC, HBE, HEK-293, CCD 841 CoN, GES-1 and L02 cells(P<0.05 or 0.01). In HEB cells cultured under hypoxic conditions, treatment with rhodiola and 3-n-butylphthalide significantly promoted the cell proliferation and improved the cell viability(P<0.05), and obviously reversed hypoxia-induced increase in LDH activity(P<0.05 or P<0.01). Conclusion Among the 7 cell lines tested in this study, HEB cells are the most sensitive to hypoxia and can serve as an ideal model for studying drug interventions of hypoxic cell injuries.
引文
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[4] LI Y, QIU L, LIU X, et al. PINK1 alleviates myocardial hypoxia-reoxygenation injury by ameliorating mitochondrial dysfunction[J]. Biochem Biophys Res Commun, 2017, 484(1): 118-124. DOI: 10.1016/j.bbrc.2017.01.061.
[5] BURTSCHER M, GATTERER H, BURTSCHER J, et al. Extreme terrestrial environments: life in thermal stress and hypoxia. A narrative review[J]. Front Physiol, 2018, 9: 572. DOI: 10.3389/fphys.2018.00572.
[6] 高伊星. 胶质瘤缺氧转录与代谢表征的生物意义和肺癌IF1表达的临床意义[D]. 重庆: 第三军医大学, 2016. GAO Y X. The biological significance of transcriptive and metabolic features in hypoxic glioblastoma cells and clinical value of IF1 expression in lung cancer[D]. Chongqing: Third Military Medical University, 2016.
[7] 王伟. 丁苯酞预处理对脑缺血再灌注海马神经元的保护作用[D]. 承德:承德医学院, 2013. WANG W. Protective effects of 3-n-butylphthalide pretreatment on hippocampal neurons of cerebral ischemia reperfusion in rats[D]. Chengde: Chengde Medical College, 2013.
[8] 黄秀景.丁苯酞后处理对局灶性脑缺血再灌注损伤大鼠内质网应激的影响[D]. 天津:天津医科大学, 2013. HUANG X J. Effect of butylphthalide after treatment on the endoplasmic reticulum stress in focal cerebral ischemia reperfusion injury in rats[D]. Tianjin: Tianjin Medical University, 2013.
[9] 刘沛, 王延晓, 葛朝明. 红景天苷对急性脑梗死患者的相关性研究[J]. 中国医学创新, 2013, 10(8): 148-150. DOI: 10.3969/j.issn.1674-4985.2013.08.092. LIU P, WANG Y X, GE C M. Correlative study of salidroside in patients with acute cerebral infarction[J]. Med Innovation China, 2013, 10(8): 148-150. DOI: 10.3969/j.issn.1674-4985.2013.08.092.