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在减轻大鼠睾丸缺血/再灌注损伤的退行性影响方面,Trolox比别嘌呤醇更有效。

Trolox is more successful than allopurinol to reduce degenerative effects of testicular ischemia/reperfusion injury in rats.

作者信息

Seker Ugur, Nergiz Yusuf, Aktas Ayfer, Akkus Murat, Ozmen Mehmet Ferit, Uyar Emre, Soker Sevda

机构信息

Department of Histology and Embryology, Faculty of Medicine, Dicle University, Turkey.

Department of Histology and Embryology, Faculty of Medicine, Dicle University, Turkey.

出版信息

J Pediatr Urol. 2020 Aug;16(4):465.e1-465.e8. doi: 10.1016/j.jpurol.2020.05.008. Epub 2020 Jun 6.

Abstract

INTRODUCTION

Reperfusion surgery following testicular ischemia is a reproductive health threatening status and may result with organ dysfunction in men. The high level of reactive oxygen species (ROS) and cease of blood flow to the testis are the most important reasons of this testicular injury. Until today, numerous experimental studies reported that antioxidants might be efficient to alleviate oxidative stress induced organ dysfunction. For this purpose, in this study, we have investigated the protective effects of xanthine oxidase (XO) inhibitor, allopurinol, and ROS scavenger, trolox, in a comparative perspective in testicular ischemia reperfusion injury subjected rats.

MATERIALS AND METHODS

Twenty-eight adult male Sprague Dawley rats were divided into four groups of seven animals in each; control, ischemia/reperfusion (I/R), allopurinol and trolox. The rats in control group did not receive any application. Animals in I/R, allopurinol and trolox groups were subjected to 2 h testicular reperfusion injury following 5 h ischemia. Intraperitoneally (i.p.) 1 ml isotonic, 200 mg/kg allopurinol and 50 mg/kg trolox were administered to the animals in these groups 30 min prior reperfusion. At the end of experiment, all animals were sacrificed and blood serum malondialdehyde (MDA) levels were measured. Histological sections were obtained from the testis and stained with hematoxylin and eosin (H&E), proliferating cell nuclear antigen (PCNA) and cleaved caspase-3. Apoptotic index was evaluated with TUNEL Assay.

RESULTS

Severe morphological degenerations, increased serum MDA, cleaved caspase-3 and TUNEL Assay positivity rate, but reduced PCNA positivity rate was observed in ischemia and reperfusion group. Morphological degenerations, MDA level, apoptotic index and PCNA positive cell rate were slightly alleviated in allopurinol administered animals compared with ischemia and reperfusion group. Protection with trolox was more successful and the results of the analysis were similar to the control group.

DISCUSSION

Ischemia that leading to testicular torsion is a reproductive health affecting problem and current surgical treatment methods might be insufficient to recover testis. Various types of ROS generating mechanisms in cell are limiting protective potency of allopurinol, and cocktail administration of different ROS inhibitors might be more effective. However, our results indicate that free radical scavenger trolox might be a candidate drug to alleviate degenerative effects of testicular ischemia reperfusion injury.

CONCLUSIONS

This is the first study that demonstrates antioxidant trolox was more successful than XO inhibitor allopurinol to protect testis against ischemia and reperfusion injury in rats.

摘要

引言

睾丸缺血后的再灌注手术会对生殖健康构成威胁,可能导致男性器官功能障碍。活性氧(ROS)水平升高和睾丸血流中断是这种睾丸损伤的最重要原因。迄今为止,众多实验研究报告称,抗氧化剂可能有效减轻氧化应激诱导的器官功能障碍。为此,在本研究中,我们从比较的角度研究了黄嘌呤氧化酶(XO)抑制剂别嘌呤醇和ROS清除剂曲洛昔芬对睾丸缺血再灌注损伤大鼠的保护作用。

材料与方法

28只成年雄性Sprague Dawley大鼠分为四组,每组7只;对照组、缺血/再灌注(I/R)组、别嘌呤醇组和曲洛昔芬组。对照组大鼠未接受任何处理。I/R组、别嘌呤醇组和曲洛昔芬组的动物在缺血5小时后接受2小时的睾丸再灌注损伤。在再灌注前30分钟,分别向这些组的动物腹腔内注射1毫升等渗溶液、200毫克/千克别嘌呤醇和50毫克/千克曲洛昔芬。实验结束时,处死所有动物并测量血清丙二醛(MDA)水平。从睾丸获取组织切片,并用苏木精和伊红(H&E)、增殖细胞核抗原(PCNA)和裂解的半胱天冬酶-3进行染色。用TUNEL法评估凋亡指数。

结果

在缺血再灌注组中观察到严重的形态学退变、血清MDA升高、裂解的半胱天冬酶-3和TUNEL法阳性率升高,但PCNA阳性率降低。与缺血再灌注组相比,给予别嘌呤醇的动物的形态学退变、MDA水平、凋亡指数和PCNA阳性细胞率略有减轻。曲洛昔芬的保护作用更成功,分析结果与对照组相似。

讨论

导致睾丸扭转的缺血是一个影响生殖健康的问题,目前的手术治疗方法可能不足以使睾丸恢复。细胞中各种类型的ROS生成机制限制了别嘌呤醇的保护效力,联合使用不同的ROS抑制剂可能更有效。然而,我们的结果表明,自由基清除剂曲洛昔芬可能是一种减轻睾丸缺血再灌注损伤退行性影响的候选药物。

结论

这是第一项表明抗氧化剂曲洛昔芬在保护大鼠睾丸免受缺血再灌注损伤方面比XO抑制剂别嘌呤醇更成功的研究。

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