| 492 | 4 | 111 |
| 下载次数 | 被引频次 | 阅读次数 |
目的 探究普罗布考对过氧化氢(H_2O2)介导的小鼠睾丸间质细胞系(TM3)损伤的影响。方法 体外培养TM3细胞，分为3组：对照组、H_2O2损伤模型组和普罗布考处理组。CCK8检测细胞存活率，流式细胞术检测细胞凋亡率，活性氧(ROS)检测试剂盒检测细胞内ROS水平，ELISA测定细胞上清睾酮水平，实时荧光定量PCR检测细胞内睾酮合成酶类固醇激素合成急性调节蛋白(StAR)和17β-羟基类固醇脱氢酶(HSD-17β)、细胞凋亡指标Bax、Caspase3和氧化应激相关指标核因子E2相关因子2 (NRF2)、血红素加氧酶-1(HO-1)的mRNA水平，Western blot检测细胞内Bax和Caspase3蛋白表达改变。结果 与对照组比较，H_2O2损伤模型组细胞存活率显著降低、凋亡率显著升高(P<0.05),Bax和Caspase3表达显著上调(P<0.05),细胞内ROS水平显著升高、NRF2和HO-1水平显著降低(P<0.05),睾酮分泌量和睾酮合成酶表达显著降低(P<0.05);与H_2O2损伤模型组比较，普罗布考处理组细胞存活率显著升高、凋亡率显著下降(P<0.05),Bax和Caspase3表达显著下调(P<0.05),细胞内ROS水平显著下降、NRF2和HO-1水平显著升高(P<0.05),睾酮分泌量和睾酮合成酶表达增高(P<0.05)。结论 普罗布考可以减轻H_2O2诱导的TM3细胞凋亡和睾酮合成异常，这种保护作用可能与普罗布考的抗氧化和抗凋亡效应有关。
Abstract：Objective:To investigate the effects of probucol on H_2O2-mediated Leydig cells(TM3) injury.Methods:TM3 cells were cultured in vitro and divided into three groups: control group, H_2O2 injury group and probucol treatment group. The cell survival rate was detected by CCK8,the cell apoptosis rate was detected by flow cytometry, and intracellular reactive oxygen species(ROS) level was determined by ROS test kit. The testosterone level in cell supernatant was detected by ELISA,and the mRNAs of intracellular testosterone synthase steroidogenic acute regulatory protein(StAR) & 17β-hydroxysteroid dehydrogenase(HSD-17β),apoptosis indicator Bax, Caspase3 and oxidative stress-related indicator nuclear factor erythroid 2-related factor 2(NRF2) and heineoxygenase-l(HO-1) were detected by real-time PCR. The protein expression of Bax and Caspase3 was detected by Western blot. Results:Compared with the control group, the cell survival rate was significantly decreased and the apoptosis rate was significantly increased(P<0.05),the expression of Bax and Caspase3 was significantly up-regulated(P<0.05),the intracellular ROS levels were significantly increased(P<0.05),the levels of NRF2 and HO-1 were significantly decreased(P<0.05),and the secretion of testosterone and the expression of testosterone synthase were significantly decreased(P<0.05) in the H_2O2 injury group. Compared with the H_2O2 injury group, the cell survival rate was significantly increased(P<0.05),the apoptosis rate was significantly decreased(P<0.05),the expression of Bax and Caspase3 was significantly down-regulated(P<0.05),intracellular ROS level was significantly decreased(P<0.05),NRF2 and HO-1 levels were significantly increased(P<0.05),and testosterone secretion and testosterone synthase expression were significantly increased(P<0.05) after probucol treatment.Conclusions:Probucol can alleviate H_2O2 induced apoptosis and abnormal testosterone synthesis of TM3 cells, and this protective effect may be related to the effect of antioxidant and anti-apoptotic effects of probucol.
[1] 陈子江，刘嘉茵，黄荷凤，等.不孕症诊断指南[J].中华妇产科杂志，2019,54:505-511.
[2] Mu Y,Dai HG,Luo LB,et al.Irisin alleviates obesity-related spermatogenesis dysfunction via the regulation of the AMPKalpha signalling pathway[J].Reprod Biol Endocrinol,2021,19:135.
[3] Fainberg J,Kashanian JA.Recent advances in understanding and managing male infertility[J].F1000Res,2019,8:F1000 Faculty Rev-670.
[4] Maresch CC,Stute DC,Alves MG,et al.Diabetes-induced hyperglycemia impairs male reproductive function:a systematic review[J].Hum Reprod Update,2018,24:86-105.
[5] Ding GL,Liu Y,Liu ME,et al.The effects of diabetes on male fertility and epigenetic regulation during spermatogenesis[J].Asian J Androl,2015,17:948-953.
[6] 夏雨果，陈秋，汤志梅，等.三才连梅颗粒对糖尿病小鼠睾丸组织Nrf2/HO-1信号通路的影响[C].首届男性大健康中西医协同创新论坛暨第三届全国中西医结合男科青年学术论坛，2019,南昌.
[7] 穆杨，杨菁.二甲双胍对H2O2诱导的TM3小鼠睾丸间质细胞氧化应激和细胞凋亡的影响[J].生殖医学杂志，2021,30:644-648.
[8] 王建平，张杰，赵洪云.普罗布考对高脂血症大鼠脑缺血再灌注的脑保护作用及机制研究[J].重庆医学，2022,51:1106-1111.
[9] 张玉静，刘倩.普罗布考对早期糖尿病肾病患者氧化应激及转化生长因子-β1的影响[J].医学信息，2021,34:112-114.
[10] Xie Y,Song A,Zhu Y,et al.Effects and mechanisms of probucol on aging-related hippocampus-dependent cognitive impairment[J].Biomed Pharmacother,2021,144:112266.
[11] 刘松涛，曹会峰，郝鹏，等.普罗布考对糖尿病大鼠生育功能的影响[J].黑龙江医药科学，2017 40:63-64.
[12] Santos DB,Colle D,Moreira ELG,et al.Probucol protects neuronal cells against peroxide-induced damage and directly activates glutathione peroxidase-1[J].Mol Neurobiol,2020,57:3245-3257.
[13] Gianatti EJ,Grossmann M.Testosterone deficiency in men with Type 2 diabetes:pathophysiology and treatment[J].Diabet Med,2020,37:174-186.
[14] Suarez AM,Israeli JM,Kresch E,et al.Testosterone therapy in children and adolescents:to whom,how,when?[J] Int J Impot Res,2022,34:652-662.
[15] Li X,Nokkala E,Yan W,et al.Altered structure and function of reproductive organs in transgenic male mice overexpressing human aromatase[J].Endocrinology,2001,142:2435-2442.
[16] 蔡旺春，胡亚楠，胡振兴，等.Nesfatin-1对高糖高脂诱导的睾丸间质细胞氧化应激的干预作用[J].宁夏医科大学学报，2020,42:372-376.
[17] Al-Majed AA.Probucol attenuates oxidative stress,energy starvation,and nitric acid production following transient forebrain ischemia in the rat hippocampus[J].Oxid Med Cell Longev,2011,2011:471590.
[18] El-Demerdash E,Awad AS,Taha RM,et al.Probucol attenuates oxidative stress and energy decline in isoproterenol-induced heart failure in rat[J].Pharmacol Res,2005,51:311-318.
[19] Xiao X,Hou H,Lin V,et al.Probucol protects rats from cardiac dysfunction induced by oxidative stress following cardiopulmonary resuscitation[J].Oxid Med Cell Longev,2017,2017:1284804.
[20] Ruan Z,Wang H,Zhang K,et al.Probucol improves erectile function by regulating endoplasmic reticulum stress in rats with streptozotocin-induced diabetes[J].Andrologia,2021,53:e13999.
[21] Wei SM,Huang YM,Zhou J.Probucol reduces testicular torsion/detorsion-induced ischemia/reperfusion injury in rats[J].Oxid Med Cell Longev,2017,2017:5424097.
[22] Fiore M,Degrassi F.Dimethyl sulfoxide restores contact inhibition-induced growth arrest and inhibits cell density-dependent apoptosis in hamster cells[J].Exp Cell Res,1999,251:102-110.
[23] 李俊东，裴晓川，白春强，等.二甲基亚砜(DMSO)对骨髓瘤细胞RPMI8226影响的研究[J].吉林医学，2017,38:83-84.
[24] 杨晓航，李舒，刘文奇，等.二甲基亚砜、二甲基酰胺、甲醇、乙醇四种溶剂对Hela细胞培养干预效应研究[J].陕西医学杂志，2016,45:774-779.
基本信息:
中图分类号:R711.6
引用信息:
[1]罗凌波,穆杨,杨菁.普罗布考对H_2O_2诱导的TM3小鼠睾丸间质细胞内氧化应激和细胞凋亡的影响[J].生殖医学杂志,2023,32(03):420-426.
基金信息:
国家自然科学基金(81971356);国家自然科学基金青年项目(82001609)
2023-03-15
2023-03-15