模拟微重力环境下大鼠原代睾丸细胞形态功能、活力和抗氧化系统的改变

V. Bonetto, L. Scarabelli, M. Masini
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引用次数: 0

摘要

研究了短期模拟微重力环境对青春期前Wistar大鼠睾丸原代细胞培养的影响。利用免疫荧光技术检测细胞骨架和性激素结合球蛋白(SHBG/ABP)的变化,利用RT-PCR检测抗氧化系统和细胞活力。细胞在三维恒温器随机定位机(RPM)上培养6和24小时。在每次实验结束时,一旦停止RPM旋转,细胞要么被固定在多聚甲醛中,要么被裂解并提取RNA。在暴露于SMG的细胞中,细胞骨架变得紊乱,微管碎片化,并且在处理6小时后已无法检测到SHBG。此外,SMG暴露24h后,各种抗氧化系统显著增加。最初,SMG似乎扰乱了允许睾丸支持精子生产的抗氧化保护策略,从而产生了一种类似衰老的氧化应激状态。在真实微重力条件下进行的短期重力变化研究,可以提供更多关于这种条件下睾丸内甾体生成和生殖细胞分化的信息,并证实模拟方法的有效性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Morphofunctional, viability and antioxidant system alterations on rat primary testicular cells exposed to simulated microgravity
This study focused on effects induced by short-term simulated microgravity (SMG) condition on primary cell culture from pre-pubertal Wistar rats testis. Cells were analyzed for cytoskeletal and Sex Hormone Binding Globulin (SHBG/ABP) changes by immunofluorescence technique, for antioxidant system exploiting RT-PCR and cell viability. Cells were cultured for 6 and 24h on a three-dimensional clinostat, Random Positioning Machine (RPM). At the end of each experiment, once stopped the RPM rotation, cells were either fixed in paraformaldehyde or lysed and RNA extracted. In cells exposed to SMG the cytoskeleton became disorganized, microtubules fragmented and SHBG was already undetectable after 6h of treatment. Moreover, various antioxidant systems significantly increased after 24h of SMG exposure. Initially, SMG seemed to disturb antioxidant protection strategies allowing the testes to support sperm production, thus generating an aging-like state of oxidative stress. Studies on changes induced by short-term altered gravity conditions, carried out in real microgravity, could give more information on steroidogenesis and germ cell differentiation within the testis exposed to this condition and confirm the validity of simulation approach.
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