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睾丸扭转及再灌注:生化和分子改变的证据。

Testicular torsion and reperfusion: evidences for biochemical and molecular alterations.

机构信息

Department of Basic Sciences, Faculty of Veterinary Medicine, Urmia University, P.O. BOX: 1177, Urmia, Iran.

Department of Surgery and Diagnostic Imaging, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran.

出版信息

Cell Stress Chaperones. 2018 May;23(3):429-439. doi: 10.1007/s12192-017-0855-0. Epub 2017 Oct 31.

Abstract

This study was done in order to determine the molecular and biochemical alterations following testicular torsion (TT) and torsion-reperfusion (TR). For this purpose, 54 male Wistar rats were divided into five groups as control group (n = 6) and experimental group subjected to 1, 2, 4, and 8 h unilateral left torsion induction (n = 12 in each group). After induction of TT, testicular samples were collected from each group (n = 6), and the other six rats of each group underwent the same period of reperfusion after TT and then were sampled. Histological changes, the mRNA and protein expression of heat shock protein-70 (Hsp70), and caspase-3 were examined using reverse transcriptase-PCR (RT-PCR) and immunohistochemistry, respectively. Testicular total antioxidant capacity (TAC), glutathione peroxidase (GSH-px), and malondialdehyde (MDA) levels were evaluated. The mRNA damage and DNA fragmentation were assessed. The TT and TR significantly reduced differentiation and spermiogenesis indices (p < 0.05). The TT- and TR-induced groups exhibited a severe reduction in Hsp70 expression as well as remarkable enhancement in caspase-3 expression. The TAC and GSH-px levels were decreased and the MDA content was increased in TT- and TR-induced groups. Finally, the TT and TR enhanced mRNA damage and DNA fragmentation. The TT- and TR-induced damaging oxidative stress, diminished Hsp70 expression, and enhanced caspase-3 mRNA and protein levels result in apoptosis following 1, 2, and 4 h. Whereas, following 8 h, TT and TR initiate the necrosis by inducing energy depletion as well as severe mRNA damage.

摘要

本研究旨在确定睾丸扭转(TT)和扭转再灌注(TR)后分子和生化的改变。为此,将 54 只雄性 Wistar 大鼠分为对照组(n=6)和实验组,实验组分别进行 1、2、4 和 8 h 的左侧单侧扭转诱导(每组 n=12)。TT 诱导后,从每组收集睾丸样本(n=6),每组的另外 6 只大鼠在 TT 后经历相同的再灌注期,然后取样。使用逆转录聚合酶链反应(RT-PCR)和免疫组织化学分别检查热休克蛋白 70(Hsp70)的 mRNA 和蛋白表达、组织学变化、丙二醛(MDA)水平、总抗氧化能力(TAC)、谷胱甘肽过氧化物酶(GSH-px)。评估 DNA 片段化和 mRNA 损伤。TT 和 TR 显著降低了分化和精子发生指数(p<0.05)。TT 和 TR 诱导组的 Hsp70 表达明显减少,caspase-3 表达明显增强。TT 和 TR 诱导组的 TAC 和 GSH-px 水平降低,MDA 含量增加。最后,TT 和 TR 增强了 mRNA 损伤和 DNA 片段化。TT 和 TR 诱导的破坏性氧化应激、Hsp70 表达减少以及 caspase-3 mRNA 和蛋白水平升高导致 1、2 和 4 h 后发生凋亡。然而,在 8 h 后,TT 和 TR 通过诱导能量耗竭和严重的 mRNA 损伤引发坏死。

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