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精索静脉曲张不育男性精子样本中作为热敏感电压依赖性离子通道的表达及氧化应激:一项病例对照研究

Expression of as A Heat Sensitive Voltage-Dependent Ion Channel and Oxidative Stress in Sperm Samples of Infertile Men with Varicocele: A Case-Control Study.

作者信息

Salahshouri Sahar, Akbarian Fahimeh, Tavalaee Marziyeh, Seifati Seyed Morteza, Nasr-Esfahani Mohammad Hossein

机构信息

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

Department of Biology , Medical Biotechnology Research Center, Ashkezar Branch, Islamic Azad University, Ashkezar, Yazd, Iran.

出版信息

Cell J. 2022 Jun;24(6):323-329. doi: 10.22074/cellj.2022.8038. Epub 2022 Jun 29.

Abstract

OBJECTIVE

Transient receptor potential vanilloid 1 (TRPV1) is a heat-activated nonselective cation channel that plays important role in the spermatogenesis, capacitation, acrosome reaction and sperm/oocyte fusion. Considering the high testicular temperature and oxidative stress in varicocele condition, we aimed to assess expression of TRPV1 in sperm of infertile men.

MATERIALS AND METHODS

In this case-control study, twenty-five men with varicocele (grade II and III) as well as twentyfive fertile were recruited. Sperm parameters, protamine deficiency (Chromomycin A3), DNA damage (TUNEL), lipid peroxidation (BODIPY), gene expression (real time polymerase chain reaction), TRPV1 protein (flowcytometry and immunocytochemical techniques), and acrosome reaction were assessed between fertile and varicocele groups.

RESULTS

We observed a significant decrease in the sperm parameters, and also, an increased DNA damage, lipid peroxidation, and protamine deficiency in varicocele group. Although, the mRNA expression of was similar between varicocele and fertile groups, its expression at the protein level was significantly decreased in the varicocele group in comparison with fertile group. Additionally, the TRPV1 localization was changed from the equatorial region to the acrosomal region of the head, especially in the acrosomal region, which was more significant in the fertile group than the varicocele group after inducing acrosome reaction.

CONCLUSION

In addition to the quality of sperm parameters, and chromatin integrity that were lower significantly in varicocele group, the expression of TRPV1 protein was also lower in varicocele condition that could be associated with reduced capacitation, acrosome reaction and sperm/oocyte fusion and thereby infertility.

摘要

目的

瞬时受体电位香草酸亚型1(TRPV1)是一种热激活的非选择性阳离子通道,在精子发生、获能、顶体反应及精卵融合过程中发挥重要作用。鉴于精索静脉曲张患者睾丸温度较高且存在氧化应激,我们旨在评估不育男性精子中TRPV1的表达情况。

材料与方法

在这项病例对照研究中,招募了25名患有精索静脉曲张(II级和III级)的男性以及25名生育能力正常的男性。对生育能力正常组和精索静脉曲张组的精子参数、鱼精蛋白缺乏情况(用放线菌素A3检测)、DNA损伤情况(用TUNEL法检测)、脂质过氧化情况(用BODIPY检测)、基因表达情况(实时聚合酶链反应)、TRPV1蛋白(流式细胞术和免疫细胞化学技术)以及顶体反应进行了评估。

结果

我们观察到精索静脉曲张组的精子参数显著降低,同时DNA损伤、脂质过氧化及鱼精蛋白缺乏情况增加。虽然精索静脉曲张组和生育能力正常组之间TRPV1的mRNA表达相似,但与生育能力正常组相比,精索静脉曲张组其蛋白水平的表达显著降低。此外,TRPV1的定位从头部的赤道区域转移到了顶体区域,尤其是在顶体区域,诱导顶体反应后,生育能力正常组的这种变化比精索静脉曲张组更明显。

结论

除了精索静脉曲张组精子参数质量和染色质完整性显著较低外,精索静脉曲张情况下TRPV1蛋白的表达也较低,这可能与获能、顶体反应及精卵融合减少从而导致不育有关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6346/9315213/1a58f1ae9c8b/Cell-J-24-323-g01.jpg

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