氯胺酮对HepG2细胞线粒体功能的影响

发布时间：2018-01-19 13:22

本文关键词： 氯胺酮 Hep G细胞线粒体药物毒性　出处：《中国药理学与毒理学杂志》2015年01期 　论文类型：期刊论文

【摘要】：目的评价氯胺酮的线粒体毒性,从而探讨氯胺酮可能的毒性机制。方法 Hep G2细胞培养基中分别加入齐多夫定10~2000μmol·L-1培养7 d,或加入氯胺酮50~3000μmol·L-1培养24 h,CCK8细胞计数法测定细胞存活率。Hep G2细胞培养基中分别加入齐多夫定100μmol·L-1培养7 d后,或加入氯胺酮100和1200μmol·L-1培养24 h,化学发光法检测胞内ATP合成水平,荧光探针法检测胞内钙离子浓度、活性氧及线粒体膜电位(ΔΨm)的变化,同时设齐多夫定1000μmol·L-1用于实时荧光定量PCR检测线粒体DNA(mt DNA)表达水平的变化。结果与正常对照组相比,齐多夫定10~2000μmol·L-1可以抑制细胞存活,IC50为100μmol·L-1;氯胺酮1000~3000μmol·L-1抑制细胞存活,IC50为1200μmol·L-1。与正常对照组相比,齐多夫定100μmol·L-1组线粒体ATP合成水平显著下降(P0.05),胞内钙离子浓度明显升高(P0.05),活性氧水平显著升高(P0.05),ΔΨm显著下降(P0.05),而1000μmol·L-1可以明显干扰mt DNA表达水平(P0.05)。氯胺酮100μmol·L-1组ATP合成水平明显下降(P0.05),其他指标均无显著性差异;氯胺酮1200μmol·L-1组ATP合成水平显著下降(P0.01),活性氧水平和胞内钙离子浓度显著升高(P0.01),ΔΨm显著下降(P0.05),mt DNA水平无明显改变。结论氯胺酮可以通过干扰线粒体代谢功能而诱导线粒体损伤。
[Abstract]:Objective to evaluate the mitochondrial toxicity of ketamine. Methods Hep G2 cell culture medium was cultured with zidovudine 102000 渭 mol 路L-1 for 7 days. Or Ketamine 50 渭 mol 路L -1 for 24 h. The survival rate of Hep G2 cells was measured by CCK8 cell count method. Zidovudine 100 渭 mol 路L -1 was added to the cell culture medium for 7 days. After incubation with ketamine 100 and 1200 渭 mol 路L -1 for 24 h, intracellular ATP synthesis was detected by chemiluminescence assay and intracellular calcium concentration was detected by fluorescence probe method. Changes of reactive oxygen species and mitochondrial membrane potential (螖蠄 m). At the same time, zidovudine 1000 渭 mol 路L -1 was used to detect the expression of mitochondrial DNA(mt by real-time fluorescence quantitative PCR. The IC50 of zidovudine was 100 渭 mol 路L ~ (-1), and that of zidovudine 10 ~ (2 000) 渭 mol 路L ~ (-1) could inhibit cell survival. The IC50 of Ketamine 1000 ~ 3000 渭 mol 路L ~ (-1) inhibited cell survival was 1 200 渭 mol 路L ~ (-1), compared with that of normal control group. The level of mitochondrial ATP synthesis in zidovudine 100 渭 mol 路L -1 group decreased significantly (P 0.05), and the intracellular calcium concentration increased significantly (P 0.05). The level of reactive oxygen species increased significantly (P 0.05), and 螖蠄 m decreased significantly (P 0.05). However, 1 000 渭 mol 路L -1 significantly interfered with the expression of Mt DNA. The ATP synthesis of ketamine 100 渭 mol 路L -1 group decreased significantly (P 0.05). . There was no significant difference in other indexes. In ketamine 1200 渭 mol 路L -1 group, the level of ATP synthesis decreased significantly, while the level of reactive oxygen species and intracellular calcium concentration increased significantly (P 0.01). 螖蠄 m significantly decreased the level of P0. 05 mt DNA. Conclusion Ketamine can induce mitochondrial damage by interfering with mitochondrial metabolic function.
【作者单位】：中国医药工业研究总院上海医药工业研究院国家上海新药安全评价研究中心;
【基金】：国家科技重大专项(2012ZX09302002);国家科技重大专项(2012ZX09505-001-003) 国家自然科学基金(81273603) 上海市青年科技启明星计划(14QB1400400)~~
【分类号】：R114
【正文快照】： 氯胺酮是临床唯一具有麻醉、镇痛及镇静等多项药效于一体的静脉麻醉药物,在现代临床医学中已被广泛应用。据美国FDA统计,氯胺酮的毒性作用主要表现在肝、神经、心及肾功能等方面[1]。因为该药具有致幻特性,又被娱乐性使用及滥用[2]。因此,控制氯胺酮的毒性作用成为临床应用及

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