• 中国中文核心期刊
  • 中国科学引文数据库核心期刊
  • 中国科技核心期刊
  • 中国高校百佳科技期刊
高级检索

耳聋左慈丸对HEI-OC1细胞氧化应激治疗作用的机制研究

卢国侠, 顾云双, 郑亦文, 彭英, 阿基业, 王广基, 孙建国

卢国侠,顾云双,郑亦文,等. 耳聋左慈丸对HEI-OC1细胞氧化应激治疗作用的机制研究[J]. 中国药科大学学报,2025,56(2):188 − 195. DOI: 10.11665/j.issn.1000-5048.2024102701
引用本文: 卢国侠,顾云双,郑亦文,等. 耳聋左慈丸对HEI-OC1细胞氧化应激治疗作用的机制研究[J]. 中国药科大学学报,2025,56(2):188 − 195. DOI: 10.11665/j.issn.1000-5048.2024102701
LU Guoxia, GU Yunshuang, ZHENG Yiwen, et al. Therapeutic mechanism of of Erlong Zuoci Pills on oxidative stress in HEI-OC1 cells[J]. J China Pharm Univ, 2025, 56(2): 188 − 195. DOI: 10.11665/j.issn.1000-5048.2024102701
Citation: LU Guoxia, GU Yunshuang, ZHENG Yiwen, et al. Therapeutic mechanism of of Erlong Zuoci Pills on oxidative stress in HEI-OC1 cells[J]. J China Pharm Univ, 2025, 56(2): 188 − 195. DOI: 10.11665/j.issn.1000-5048.2024102701

耳聋左慈丸对HEI-OC1细胞氧化应激治疗作用的机制研究

基金项目: 

undefined

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)

详细信息
    通讯作者:

    孙建国: Tel:13915990907 E-mail:jgsun@cpu.edu.cn

    #卢国侠与顾云双为共同第一作者

  • 中图分类号: R965

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

Funds: 

This study was supported by the National Key Research and Development Program for Intergovernmental Cooperation in International Science and Technology Innovation (2017YFE0109600)

undefined

  • 摘要:

    探究耳聋左慈丸对过氧化氢(H2O2)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,0.9 mmol/L的H2O2诱导12 h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H2O2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H2O2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。

    Abstract:

    To the present study aimed to investigate the protective effects of Erlong Zuoci Pills on oxidative stress induced by hydrogen peroxide (H2O2) 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 H2O2 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 H2O2. 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 H2O2-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.

  • Figure  1.   Proliferation rate of House Ear Institute-Organ of Corti 1 (HEI-OC1) cells under different concentrations of hydrogen peroxide ($\bar{x}\pm s $,n=5)

    Figure  2.   Pharmacodynamic indexes ($\bar{x}\pm s $,n=5)

    A: Cell viability of HEI-OC1 cells; B: Different drugs on lactate dehydrogenase(LDH)release amounts in HEI-OC1 cells supernatant; C: Different drugs on reactive oxygen species (ROS)content in HEI-OC1 cells; D: Different drugs on superoxide dismutase (SOD)activity in HEI-OC1 cells. (Control: Control group; Model: Model group; TQH, TQM,TQL: High, medium, and low dose groups of ELZC; VC: Ascorbic acid group) ***P<0.001 vs control group; #P<0.05,##P<0.01,###P<0.001 vs model group

    Figure  3.   Multivariate statistical analysis plots($\bar{x}\pm s $,n=5)

    A: Principal component analysis plot; B: Partial least squares discriminant analysis plot; C: Permutation test

    Figure  4.   Differential metabolite screening analysis($\bar{x}\pm s $,n=5)

    A: Volcano plot of all tested substances between Control group and Model group; B: Orthogonal Partial Least Squares Discriminant Analysis (OPLS-DA) score chart between Control group and Model group; C: All tested substances between Control group and Model group are sorted by Variable Importance in Projection (VIP)value; D: Volcano plot of all tested substances between Model group and TQM group; E: OPLS-DA score chart between Model group and TQM group; F: All tested substances between Model group and TQM group are sorted by VIP value

    Figure  5.   Metabolomics analysis($\bar{x}\pm s $,n=5)

    A: Heat map; B: Metabolic pathway analysis diagram

  • [1]

    Baguley D, McFerran D, Hall D. Tinnitus[J]. Lancet, 2013, 382(9904): 1600-1607. doi: 10.1016/S0140-6736(13)60142-7

    [2]

    Bauer CA. Tinnitus[J]. N Engl J Med, 2018, 378(13): 1224-1231. doi: 10.1056/NEJMcp1506631

    [3]

    Cheng YF, Xirasagar S, Kuo NW, et al. Tinnitus and risk of attempted suicide: a one year follow-up study[J]. J Affect Disord, 2023, 322: 141-145. doi: 10.1016/j.jad.2022.11.009

    [4] LI L, YAO J, LIU LF, et al. Research status of chronic tinnitus patients complicated with anxiety and depression[J]. Medical Innovation of China(中国医学创新), 2023, 20(22): 181-185.
    [5]

    Sonbay ND, Saka C, Oktay Arslan B, et al. The effect of hypoxia on hearing function[J]. Turk J Med Sci, 2019, 49(5): 1450-1454. doi: 10.3906/sag-1902-210

    [6]

    Liebenberg A, Oosthuizen J, Reed S. A current affair: worker perceptions of noise exposure and occupational hearing loss in Australian coal mines[J]. Ann Work Expo Health, 2023, 67(9): 1111-1120. doi: 10.1093/annweh/wxad055

    [7]

    Chirtes F, Albu S. Prevention and restoration of hearing loss associated with the use of cisplatin[J]. Biomed Res Int, 2014, 2014: 925485.

    [8]

    Zhang WB, Ruan J, Zhang RX, et al. Age-Related Hearing Loss with tinnitus and physical frailty influence the overall and domain-specific quality of life of Chinese community-dwelling older adults[J]. Front Med (Lausanne), 2021, 8: 762556.

    [9]

    Dorobisz K, Pazdro-Zastawny K, Misiak P, et al. Sensorineural hearing loss in patients with long-COVID-19: objective and behavioral audiometric findings[J]. Infect Drug Resist, 2023, 16: 1931-1939. doi: 10.2147/IDR.S398126

    [10]

    Nawaz MU, Vinayak S, Rivera E, et al. Association between hypertension and hearing loss[J]. Cureus, 2021, 13(9): e18025.

    [11] Yan AJ, Yu YY. Clinical study on Erlong zuoci pills combined with sound therapy for chronic tinnitus of kidney essence depletion type[J]. New Chin Med (新中医), 2023, 55(3): 151-154.
    [12] Zheng P, Wang ZH. Acupuncture combined with Erlong zuoci pill in the treatment of senile tinnitus for 25 cases[J]. Chin Med Mod Distance Edu China (中国中医药现代远程教育), 2020, 18(17): 100-101.
    [13] Ge ZH, Li F, Li GB, et al. Clinical study on Erlong zuoci pill combined with water acupuncture in treatment of senile sudden deafness with kidney Yin deficiency type[J]. Mod Chin Med (现代中医药), 2022, 42(5): 111-115.
    [14] Zou LY. Clinical study on Erlong zuoci pills for presbycusis[J]. J New Chin Med (新中医), 2020, 52(23): 25-27.
    [15] Ping Y, Zhang Q, Feng XM. Effects of verbascoside on hypoxia/reoxygenation induced cardiomyocyte injury[J]. Chin J Integr Med Cardio Cerebrovasc Dis (中西医结合心脑血管病杂志), 2022, 20(20): 3698-3703.
    [16] Wu LY, Wu TT, Li YL, et al. A model of H2O2-induced oxidative stress damage in hair cell-like HEI-OC1 cells[J]. Chin J Otol (中华耳科学杂志), 2023, 21(4): 514-521.
    [17] Dong JF, Lu L, Lin CY, et al. Graphene inhibits damage to hair cells by oxidative stress[J]. Chin J Otol (中华耳科学杂志), 2021, 19(1): 101-107.
    [18]

    Gęgotek A, Skrzydlewska E. Ascorbic acid as antioxidant[J]. Vitam Horm, 2023, 121: 247-270.

    [19]

    Wishart DS. Emerging applications of metabolomics in drug discovery and precision medicine[J]. Nat Rev Drug Discov, 2016, 15(7): 473-484. doi: 10.1038/nrd.2016.32

    [20] Li WW, Yang Y, Wang SC, et al. Research progress of cellular metabolomics[J]. J Nanjing Univ Tradit Chin Med (南京中医药大学学报), 2017, 33(2): 187-192.
    [21] Gu YS, Wang R, Su N, et al. Pharmacokinetic study of Erlong zuoci pill in rats[J]. J China Pharm Univ (中国药科大学学报), 2022, 53(4): 481-489.
    [22] He MZ, Peng Y, Wang GJ, et al. Mechanism of sodium salicylate-induced damage to HEI-OC1 cells based on metabonomics[J]. J China Pharm Univ (中国药科大学学报), 2021, 52(5): 566-572.
    [23]

    Lu ZY, Li SJ, Sun RB, et al. Hirsutella sinensis treatment shows protective effects on renal injury and metabolic modulation in db/db mice[J]. Evid Based Complement Alternat Med, 2019, 2019: 4732858.

    [24]

    Sies H. Oxidative stress: a concept in redox biology and medicine[J]. Redox Biol, 2015, 4: 180-183. doi: 10.1016/j.redox.2015.01.002

    [25]

    Zhang BG, Huang CH, Xu DM, et al. Patulin induces ROS-dependent cardiac cell toxicity by inducing DNA damage and activating endoplasmic reticulum stress apoptotic pathway[J]. Ecotoxicol Environ Saf, 2024, 269: 115784. doi: 10.1016/j.ecoenv.2023.115784

    [26]

    Liu ZP, Li W, Geng LL, et al. Cross-species metabolomic analysis identifies uridine as a potent regeneration promoting factor[J]. Cell Discov, 2022, 8(1): 6. doi: 10.1038/s41421-021-00361-3

    [27]

    Jiang N, Zhao Z. Intestinal aging is alleviated by uridine via regulating inflammation and oxidative stress in vivo and in vitro[J]. Cell Cycle, 2022, 21(14): 1519-1531. doi: 10.1080/15384101.2022.2055252

    [28]

    Adant I, Bird M, Decru B, et al. Pyruvate and uridine rescue the metabolic profile of OXPHOS dysfunction[J]. Mol Metab, 2022, 63: 101537. doi: 10.1016/j.molmet.2022.101537

    [29]

    Al N, Çakir A, Koç C, et al. Antioxidative effects of uridine in a neonatal rat model of hyperoxic brain injury[J]. Turk J Med Sci, 2020, 50(8): 2059-2066. doi: 10.3906/sag-2002-14

    [30]

    Mironova GD, Khrenov MO, Talanov EY, et al. The role of mitochondrial KATP channel in anti-inflammatory effects of uridine in endotoxemic mice[J]. Arch Biochem Biophys, 2018, 654: 70-76. doi: 10.1016/j.abb.2018.07.006

    [31]

    Paczkowska K, Rachoń D, Berg A, et al. Alteration of branched-chain and aromatic amino acid profile as a novel approach in studying polycystic ovary syndrome pathogenesis[J]. Nutrients, 2023, 15(19): 4153. doi: 10.3390/nu15194153

    [32]

    Lynch JH, Dudareva N. Aromatic amino acids: a complex network ripe for future exploration[J]. Trends Plant Sci, 2020, 25(7): 670-681. doi: 10.1016/j.tplants.2020.02.005

    [33]

    Chen JJ, Xie J, Li WW, et al. Age-specific urinary metabolite signatures and functions in patients with major depressive disorder[J]. Aging, 2019, 11(17): 6626-6637. doi: 10.18632/aging.102133

    [34]

    di Marino S, Viceconte N, Lembo A, et al. Early metabolic response to acute myocardial ischemia in patients undergoing elective coronary angioplasty[J]. Open Heart, 2018, 5(1): e000709. doi: 10.1136/openhrt-2017-000709

    [35]

    Zhao SG, Khoo S, Ng SC, et al. Brain functional network and amino acid metabolism association in females with subclinical depression[J]. Int J Environ Res Public Health, 2022, 19(6): 3321. doi: 10.3390/ijerph19063321

    [36]

    Xie C, Wang P, Chang JW, et al. Effect of amino acids on folates accumulation in wheat seedlings during germination under red light radiation[J]. Molecules, 2022, 27(20): 6868. doi: 10.3390/molecules27206868

    [37] Wei ZY, Wang HR, Pan XH, et al. Effects of feeding regimen and dietary tryptophan level on immune function and antioxidant indices of Yangzhou geese[J]. Chin J Anim Nutr (动物营养学报), 2012, 24(12): 2356-2365.
    [38]

    Li QF, Ouyang JX, Deng CX, et al. Effects of dietary tryptophan supplementation on rectal temperature, humoral immunity, and cecal microflora composition of heat-stressed broilers[J]. Front Vet Sci, 2023, 10: 1247260. doi: 10.3389/fvets.2023.1247260

    [39] Li HW, Zhu Q, Wu LY, et al. Physiological function and dietary application of tryptophan in livestock and poultry[J]. Chin J Anim Nutr (动物营养学报), 2016, 28(3): 659-664.
    [40]

    Moro J, Tomé D, Schmidely P, et al. Histidine: a systematic review on metabolism and physiological effects in human and different animal species[J]. Nutrients, 2020, 12(5): 1414. doi: 10.3390/nu12051414

    [41]

    Sasahara I, Fujimura N, Nozawa Y, et al. The effect of histidine on mental fatigue and cognitive performance in subjects with high fatigue and sleep disruption scores[J]. Physiol Behav, 2015, 147: 238-244. doi: 10.1016/j.physbeh.2015.04.042

    [42]

    Chen Z, Li WD, Zhu LJ, et al. Effects of histidine, a precursor of histamine, on pentylenetetrazole-induced seizures in rats[J]. Acta Pharmacol Sin, 2002, 23(4): 361-366.

图(5)
计量
  • 文章访问数:  78
  • HTML全文浏览量:  22
  • PDF下载量:  37
  • 被引次数: 0
出版历程
  • 收稿日期:  2024-10-26
  • 刊出日期:  2025-04-24

目录

    /

    返回文章
    返回
    x 关闭 永久关闭