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精索静脉曲张不育男性与正常生育男性精子中自噬和凋亡途径相关主要分子标志物的比较

Comparison of main molecular markers involved in autophagy and apoptosis pathways between spermatozoa of infertile men with varicocele and fertile individuals.

作者信息

Foroozan-Broojeni Shaghayegh, Tavalaee Marziyeh, Lockshin Richard A, Zakeri Zahra, Abbasi Homayon, Nasr-Esfahani Mohammad Hossein

机构信息

Department of Reproductive Biotechnology, Reproductive Biomedicine Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran.

Department of Biological Sciences, St. John's University, Jamaica, New York.

出版信息

Andrologia. 2019 Mar;51(2):e13177. doi: 10.1111/and.13177. Epub 2018 Oct 23.

Abstract

Abnormal dilatation and tortuosity of the pampiniform plexus within the spermatic cord are termed varicocele which leads to impaired spermatogenesis due to heat-related oxidative stress and cell death. Previously, it was shown that both apoptosis and autophagy pathways were activated by heat in germ cells of mouse in vivo and in vitro. But, status of these pathways is not clear in chronic state of heat stress such as varicocele. Therefore, we aimed to access sperm apoptotic markers (active caspases 3/7 and DNA fragmentation), and autophagic markers (Atg7 and LC3 proteins) as primary outcomes, and also sperm parameters and protamine deficiency as secondary outcomes between 23 infertile men with varicocele and 16 fertile individuals. Sperm parameters were assessed according to World Health Organization 2010 protocol. Apoptotic markers (active caspases 3/7 and DNA fragmentation), autophagic markers (Atg7 and LC3 proteins), and protamine deficiency were evaluated by flow cytometry, fluorescence microscope, and western blotting techniques. Mean of autophagy and apoptosis markers, and also protamine deficiency have significantly increased in infertile men with varicocele compared to fertile individuals, but autophagy and apoptosis markers did not significantly correlate with each other. In conclusion, it seems that both apoptosis and autophagy pathways are independently active in spermatozoa of infertile men with varicocele.

摘要

精索内蔓状静脉丛的异常扩张和迂曲被称为精索静脉曲张,它会因热相关的氧化应激和细胞死亡导致精子发生受损。此前研究表明,在体内和体外实验中,热均可激活小鼠生殖细胞中的凋亡和自噬途径。但是,在精索静脉曲张这种慢性热应激状态下,这些途径的状况尚不清楚。因此,我们旨在将精子凋亡标志物(活化的半胱天冬酶3/7和DNA片段化)和自噬标志物(Atg7和LC3蛋白)作为主要观察指标,同时将精子参数和鱼精蛋白缺乏作为次要观察指标,对23名精索静脉曲张不育男性和16名生育能力正常个体进行研究。精子参数根据世界卫生组织2010年的方案进行评估。凋亡标志物(活化的半胱天冬酶3/7和DNA片段化)、自噬标志物(Atg7和LC3蛋白)以及鱼精蛋白缺乏情况通过流式细胞术、荧光显微镜和蛋白质印迹技术进行评估。与生育能力正常个体相比,精索静脉曲张不育男性的自噬和凋亡标志物平均值以及鱼精蛋白缺乏情况均显著增加,但自噬和凋亡标志物之间无显著相关性。总之,精索静脉曲张不育男性精子中的凋亡和自噬途径似乎均独立激活。

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