耳聋左慈丸对HEI-OC1细胞氧化应激治疗作用的机制研究  

Therapeutic mechanism of of Erlong Zuoci Pills on oxidative stress in HEI-OC1 cells

作　　者：卢国侠 顾云双 郑亦文 彭英[1] 阿基业[1] 王广基[1] 孙建国[1] LU Guoxia;GU Yunshuang;ZHENG Yiwen;PENG Ying;A Jiye;WANG Guangji;SUN Jianguo(Key Laboratory of Drug Metabolism and Pharmacokinetics,China Pharmaceutical University,Nanjing 210009,China;Department of Pharmacology and Toxicology,School of Biomedical Sciences,University of Otago,Dunedin 9054,New Zealand)

机构地区：[1]中国药科大学药物代谢动力学重点实验室,南京210009 [2]新西兰奥塔哥大学生物医学学院药理毒理学系,达尼丁9054

出　　处：《中国药科大学学报》2025年第2期188-195,共8页Journal of China Pharmaceutical University

基　　金：国家重点研发计划“政府间国际科技创新合作”重点专项资助项目(2017YFE0109600)。

摘　　要：探究耳聋左慈丸对过氧化氢(H_(2)O_(2))致小鼠耳蜗毛细胞株HEI-OC1氧化应激的保护作用并利用细胞代谢组学的手段研究作用机制。实验具体分组如下:对照组、模型组、耳聋左慈丸提取液低、中、高剂量组及阳性对照抗坏血酸组。以HEI-OC1细胞为研究对象,0.9mmol/L的H_(2)O_(2)诱导12h为条件进行造模。CCK-8试剂盒用于检测不同状态下HEI-OC1细胞的增殖能力;乳酸脱氢酶(LDH)、活性氧(ROS)和超氧化物歧化酶(SOD)试剂盒分别对HEI-OC1细胞中对应物质的活性和含量进行检测;并从代谢组学的角度分析了细胞内源性代谢物的变化。与模型组相比,耳聋左慈丸给药后可以显著提高细胞的增殖率,而且能改善由H_(2)O_(2)造模带来的ROS和LDH的含量升高,抗氧化酶SOD活性下降的情况。代谢组学结果显示,各组之间的偏最小二乘法判别分析得分图差异明显,并且耳聋左慈丸组有将模型组回调至空白组的趋势。耳聋左慈丸对HEI-OC1细胞氧化应激的代谢调控主要影响核苷酸代谢和氨基酸代谢。综上所述,耳聋左慈丸可以对H_(2)O_(2)致HEI-OC1细胞氧化应激起到一定的保护作用,具有抗氧化的药理活性,这种保护作用可能是增加尿苷和苯丙氨酸等氨基酸的含量进而调节嘧啶代谢、苯丙氨酸代谢、苯丙氨酸、酪氨酸和色氨酸生物合成以及组氨酸代谢等通路所产生的。To the present study aimed to investigate the protective effects of Erlong Zuoci Pills on oxidative stress induced by hydrogen peroxide(H_(2)O_(2))in House Ear Institute-Organ of Corti 1(HEI-OC1)and to explore the mechanism by cellular metabolomics.There were 6 groups in the experiment:the control group,model group,three dose groups of ELZC(low,medium,and high),and positive control ascorbic acid group.The oxidative stress injury model was established in the HEI-OC1 by inducing 0.9 mmol/L H_(2)O_(2) for 12 h.The proliferation of HEI-OC1 cells was observed by CCK-8 assay;the contents and activity of lactate hydrogenase(LDH),reactive oxygen species(ROS),and superoxide dismutase(SOD)in HEI-OC1 cells were detected by corresponding kits.Finally,the endogenous substances of cells were analyzed from the perspective of metabolomics.Compared with the model group,ELZC groups could significantly increase the cell proliferation rate after administration.Moreover,they could also ameliorate the increase of ROS and LDH content and the decrease of antioxidant enzyme SOD caused by H_(2)O_(2).Metabolomic results revealed significant differences among multiple groups in the scores of partial least squares discriminant analysis.The ELZC group could relocate the model group back to the control group.The metabolic regulation of ELZC on oxidative stress in HEI-OC1 cells mainly affects nucleotide metabolism and amino acid metabolism.In summary,the results indicate that ELZC exhibits protective effects on H_(2)O_(2)-induced oxidative stress in HEI-OC1 cells.Additionally,this protective effect may be produced by increasing the content of amino acids such as uridine and phenylalanine,thereby regulating pathways such as pyrimidine metabolism,phenylalanine metabolism,biosynthesis of phenylalanine,tyrosine,and tryptophan,and histidine metabolism.

关 键 词：耳聋左慈丸 小鼠耳蜗毛细胞株HEI-OC1 过氧化氢 氧化应激 代谢组学 

分 类 号：R965[医药卫生—药理学]