Ahmadi Kimia, Asgharzadeh Fatemeh, Mohammadpour-Asl Shadi, Ayari Fatemeh, Rahbar Fatemeh, Motazakker Morteza, Roshan-Milani Shiva, Fard Amin Abdollahzade
Student Research Committee, Urmia University of Medical Sciences, Urmia, Iran.
Department of Physiology, School of Medicine, Urmia University of Medical Sciences, Urmia, Iran.
Endocr Metab Immune Disord Drug Targets. 2023;23(13):1611-1620. doi: 10.2174/1871530323666230322092046.
Global rise in cannabis abuse during reproductive years has placed a large number of men at risk for the adverse consequences of δ-9-tetrahydrocannabinol (THC), the primary active component of cannabis. It has been reported that THC affects male fertility and causes testicular cell dysfunction and apoptosis. This study aimed to investigate the possible protective role of zinc pretreatment against the toxic effects of THC in cultured mouse Sertoli cells and the underlying mechanism.
The Mus Musculus Sertoli cell line (TM4) was cultured, exposed to THC alone (470 μM, 24 h), co-administered with zinc (8 μM, 48 h), and investigated in three groups: control, THC, and THC + zinc. The MTT was performed to evaluate cell viability. TUNEL assay was also applied for the detection of cell apoptosis and a western blot was performed for measuring protein expression levels of Caspase3, Pro-caspase3, SOD, and PDGF-A.
THC significantly decreased cell viability (p < 0.001) and expression levels of SOD, PDGF-A, and pro-caspase3 proteins (p < 0.05 for all), whereas increased Sertoli cells apoptosis (p < 0.001) and expression level of cleaved caspase3 protein (p < 0.001). Pretreatment with zinc reversed THC-induced apoptotic and oxidative effects and reduced cleaved caspase3/pro-caspase3 ratio but could not reverse THC-induced reduction of PDGF-A expression level in TM4 cells.
The present data suggest that THC induces Sertoli cell damage through a multitarget mechanism. Zinc was reported to protect against THC-induced Sertoli cell damage due to its antiapoptotic and antioxidant activities, indicating its clinical importance against THC-induced testicular toxicity among addicted men.
生育年龄期间全球大麻滥用现象增多，使大量男性面临大麻主要活性成分δ-9-四氢大麻酚（THC）产生不良后果的风险。据报道，THC会影响男性生育能力，并导致睾丸细胞功能障碍和凋亡。本研究旨在探讨锌预处理对培养的小鼠支持细胞中THC毒性作用的可能保护作用及其潜在机制。
培养小家鼠支持细胞系（TM4），分别单独暴露于THC（470μM，24小时）、与锌共同给药（8μM，48小时），分为三组进行研究：对照组、THC组和THC +锌组。采用MTT法评估细胞活力。还应用TUNEL法检测细胞凋亡，并进行蛋白质印迹法检测Caspase3、Pro-caspase3、SOD和PDGF-A的蛋白质表达水平。
THC显著降低细胞活力（p < 0.001）以及SOD、PDGF-A和Pro-caspase3蛋白的表达水平（均p < 0.05），而增加支持细胞凋亡（p < 0.001）和裂解的caspase3蛋白的表达水平（p < 0.001）。锌预处理可逆转THC诱导的凋亡和氧化作用，并降低裂解的caspase3/pro-caspase3比值，但不能逆转THC诱导的TM4细胞中PDGF-A表达水平的降低。
目前的数据表明，THC通过多靶点机制诱导支持细胞损伤。据报道，锌因其抗凋亡和抗氧化活性可保护支持细胞免受THC诱导的损伤，表明其对成瘾男性中THC诱导的睾丸毒性具有临床重要性。
