目的 研究茶黄素-3,3'-双没食子酸酯（TFDG）对衰老小鼠睾丸生精功能的改善作用并探讨其作用机制。方法 将60只SPF级雄性ICR小鼠随机分为空白对照组,溶剂对照组,衰老模型组、TFDG低、中、高剂量组[15、30、60 mg/（kg·bw）]。TFDG组和衰老模型组均采用颈背部皮下注射D-半乳糖[120 mg/（kg·bw）,连续56 d]建立衰老模型,对照组皮下注射等容量生理盐水。TFDG组同时给予不同剂量连续28 d（造模第29日开始）的TFDG灌胃处理,溶剂对照组灌胃等容量溶剂0.5% DMSO。造模/干预结束,测试小鼠睾丸生精能力,检测睾丸组织谷胱甘肽过氧化物酶（GSH-Px）、超氧化物歧化酶（SOD）、总抗氧化能力（T-AOC）,丙二醛（MDA）、葡萄糖-6-磷酸脱氢酶（G6PDH）、酸性磷酸酶（ACP）、乳酸脱氢酶（LDH）和琥珀酸脱氢酶（SDH）,分析确定抗氧化基因（Oxr1,Nrf2,Prx2）的表达。结果 与溶剂对照组比较,D-半乳糖注射降低了睾丸脏器系数、每日精子生成量和睾丸组织睾酮水平,降低了睾丸组织G6PDH、ACP、LDH、SDH、GSH-Px和SOD活性及T-AOC水平,增加了MDA含量,下调了睾丸组织Oxr1,Nrf2和Prx2的mRNA表达水平。与衰老模型组相比,TFDG干预小鼠睾丸生精功能和睾酮水平显著提高,睾丸组织G6PDH、ACP、LDH、SDH、GSH-Px和SOD活性显著增强,T-AOC水平明显升高,MDA含量显著下降,抗氧化基因mRNA表达水平显著上调。结论 在D-半乳糖所致衰老模型小鼠中,TFDG可通过提升睾丸组织抗氧化能力,上调睾丸标志酶活性和睾酮合成代谢,改善睾丸生精功能障碍。
Abstract
Objective To study the improving effect of theaflavin -3,3' -digalate (TFDG) on spermatogenesis in aging mice and explore its mechanism. Methods Sixty male ICR mice were randomly divided into blank control group, solvent control group, aging model group, TFDG low, medium and high dose groups (15, 30, 60 mg/(kg·bw)). The TFDG groups and the aging model group were given subcutaneous injection of D-galactose (120 (mg/kg·bw) for 56 consecutive days) in the region between neck and back to establish the aging model. The control group was given subcutaneous injection of normal saline of equal volume. Meanwhile, TFDG groups were given different doses of TFDG for 28 consecutive days (starting from the 29th day of modeling), and the solvent control group was given an equal volume of 0.5% DMSO. At the end of experiment, the spermatogenic function of testis was tested, and glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), total antioxidant capacity (T-AOC), malondialdehyde (MDA), glucose-6-phosphate dehydrogenase (G6PDH), acid phosphatase (ACP), lactate dehydrogenase (LDH) and succinate dehydrogenase (SDH) were measured in testicular tissues.The expression of antioxidant genes (Oxr1, Nrf2, Prx2) was determined. Results Compared with the solvent control group, D-galactose injection decreased the testis organ coefficient, daily sperm production and testosterone level. The testis G6PDH, ACP, LDH, SDH, GSH-Px, SOD activities and T-AOC level were decreased, and the MDA content increased. The mRNA expression levels of Oxr1, Nrf2 and Prx2 in testis tissue were down-regulated. Compared with the aging model group, the spermatogenic function and testosterone level of testicle were significantly improved after TFDG intervention. The activities of G6PDH, ACP, LDH, SDH, GSH-Px and SOD in testicle tissues, as well as the T-AOC level were also significantly increased.MDA content was significantly decreased. The mRNA expression level of antioxidant genes was significantly upregulated in response to TFDG intervention. Conclusion In D-galactose-induced aging mice, TFDG can improve testicular spermatogenic function by increasing the antioxidant capacity of testicular tissue, the activities of testicular enzymes and testosterone anabolism.
关键词
茶黄素-3 /
3'-双没食子酸酯 /
衰老模型 /
生精 /
抗氧化酶 /
抗氧化基因
Key words
theaflavin-3 /
3'-digallate /
models of aging /
spermatogenic function /
antioxidant enzyme /
antioxidant genes
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] Chen LK.Urbanization and population aging: Converging trends of demographic transitions in modern world[J]. Arch Gerontol Geriatr, 2022, 101: 104709.
[2] 国家统计局. 第七次全国人口普查公报(第二号): 中国[EB/OL].(2021-05-11)[2023-05-23].http://www.stats.gov.cn/xxgk/sjfb/zxfb2020/202105/t20210511_1817197.Html.
[3] Ros M, Carrascosa JM.Current nutritional and pharma-cological anti-aging interventions[J]. Biochim Biophys Acta, 2020, 1866: 165612.
[4] Wu J, Liu Y, Song Y, et al. Aging conundrum: a perspective for ovarian aging[J]. Front Endocrinol (Lausanne), 2022, 13: 952471.
[5] Santiago J, Silva J V, Alves M G, et al. Testicular aging: an overview of ultrastructural, cellular, and molecular alterations[J]. J Gerontol A Biol Sci Med Sci, 2019, 74: 860–871.
[6] 刘建亚, 冯文静, 王仁萍, 等. D-半乳糖致衰老动物模型及其机制研究进展[J]. 中华老年多器官疾病杂志, 2018, 17: 224–227.
[7] 王喆, 陆承荣, 罗渊, 等. 注射D-半乳糖致大鼠衰老效应的评价[J]. 中国医药导报, 2013, 10: 25–26,36.
[8] 宁港, 吴悔, 李波男, 等. D-半乳糖诱导迟发性性腺功能减退大鼠模型的建立与评价[J]. 中华男科学杂志, 2022, 28: 675–684.
[9] 邱竹, 姜蓉, 汪子铃, 等. 当归多糖对D-半乳糖致衰老小鼠睾丸的保护作用[J].解剖学报, 2019, 50: 506–511.
[10] Alam M, Ali S, Ashraf GM, et al. Epigallocatechin 3-gallate: from green tea to cancer therapeutics[J]. Food Chem, 2022,379: 132135.
[11] Leung LK, Su Y, Chen R, et al. Theaflavins in black tea and catechins in green tea are equally effective antioxidants[J].J Nutr, 2001, 131: 2248–2251.
[12] He HF.Research progress on theaflavins: efficacy, formation, and preparation[J]. Nutr Res, 2017, 61:1344521.
[13] 刘舒瑜,陈亚,李福轮,等.芹菜素对小鼠生精细胞周期影响的实验研究[J].环境与健康杂志,2010,27:33–35.
[14] Korkut Celikates B, Kilic V, Atli-Eklioglu O, et al. Effects of quetiapine administration on sperm quality and testicular histology[J]. Drug Chem Toxicol, 2022, 45: 2379–2387.
[15] Hu Y, Asano K, Kim S, et al. Relationship between serum testosterone and activities of testicular enzymes after continuous and intermittent training in male rats[J]. Int J Sports Med, 2004, 25: 99–102.
[16] Luo J, Mills K, le Cessie S, et al. Ageing, age-related diseases and oxidative stress: what to do next?[J]. Ageing Res Rev, 2020, 57: 100982.
[17] Jacob D, Hooten NN, Trzeciak AR, et al. Markers of oxidant stress that are clinically relevant in aging and age-related disease[J]. Mech Ageing Dev, 2013, 134: 139–157.
[18] Umbayev B, Askarova S, Almabayeva A, et al. Galactose-induced skin aging: the role of oxidative stress[J]. Oxid Med Cell Longev, 2020, 2020:7145656.
[19] Feng Y, Huang Q.Protective effects of cordycepin against d-galactose-induced aging in rats: A view from the heart[J]. Geriatr Gerontol Int, 2022, 22: 433–440.
[20] Takeshita S, Inoue N, Ueyama T, et al. Shear stress enhances glutathione peroxidase expression in endothelial cells[J]. Biochem Biophys Res Commun, 2000, 273: 66–71.
[21] Rajput SA, Shaukat A, Rajput IR, et al. Ginsenoside Rb1 prevents deoxynivalenol-induced immune injury via alleviating oxidative stress and apoptosis in mice[J]. Ecotoxicol Environ Saf, 2021, 220: 112333.
[22] Yang M, Luna L, Sørbø J G, et al. Human OXR1 maintains mitochondrial DNA integrity and counteracts hydrogen peroxide-induced oxidative stress by regulating antioxidant pathways involving p21[J]. Free Radic Biol Med, 2014, 77: 41–48.
[23] Yang M, Lin X, Rowe A, et al. Transcriptome analysis of human OXR1 depleted cells reveals its role in regulating the p53 signaling pathway[J]. Sci Rep, 2015, 5: 17409.
[24] He F, Ru X, Wen T.NRF2, a transcription factor for stress response and beyond[J]. Int J Mol Sci, 2020, 21: 4777.
[25] Rocha S, Gomes D, Lima M, et al. Peroxiredoxin 2, glutathione peroxidase, and catalase in the cytosol and membrane of erythrocytes under H2O2-induced oxidative stress[J]. Free Radic Res, 2015, 49: 990–1003.
[26] Duan T, Fan K, Chen S, et al. Role of peroxiredoxin 2 in H2O2-induced oxidative stress of primary Leydig cells[J]. Mol Med Rep, 2016, 13: 4807–4813.
[27] 谢虹, 罗志聪, 李熙灿. 茶黄素抗氧化化学机制研究[J]. 食品与机械, 2018, 34: 23–26.
[28] Su YL, Xu JZ, Ng CH, et al. Antioxidant activity of tea theaflavins and methylated catechins in canola oil [J]. J Am Oil Chem Soc, 2004, 81: 269–274.
[29] Wu Y, Li W, Xu Y, et al. Evaluation of the antioxidant effects of four main theaflavin derivatives through chemiluminescence and DNA damage analyses[J]. J Zhejiang Univ Sci B, 2011, 12: 744.
[30] Shan Z, Nisar MF, Li M, et al. Theaflavin chemistry and its health benefits[J]. Oxid Med Cell Longev, 2021, 2021:6256618.
[31] Xu C, Ni S, Xu N, et al. Theaflavin-3, 3-digallate inhibits erastin-induced chondrocytes ferroptosis via the Nrf2/GPX4 signaling pathway in osteoarthritis[J]. Oxid Med Cell Longev,2022,2022:3531995.
基金
陕西省资源生物重点实验室后补助项目(No.2015SZS-15-04); 陕西省科技重点研发计划项目（No.2021NY-121）; 陕西理工大学秦巴生物资源与生态环境省部共建国家重点实验室专项（No.SXZC-2102）; 国家自然科学基金资助项目（No.81773463）; 江苏省研究生实践创新计划（SJCX23_1703）
