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使用iTRAQ技术鉴定度他雄胺治疗的不育大鼠中参与精子成熟的新附睾管腔液蛋白

Identification of New Epididymal Luminal Fluid Proteins Involved in Sperm Maturation in Infertile Rats Treated by Dutasteride Using iTRAQ.

作者信息

Xie Shu-Wu, Li Guo-Ting, Qu Li-Juan, Cao Yang, Wang Qian, Zhou Jie-Yun, Zhong Rui-Hua, Guo Xiang-Jie, Zhu Yan

机构信息

Department of Reproductive Pharmacology, Key Lab of Reproduction Regulation of NPFPC, SIPPR, IRD, Fudan University, Shanghai 200032, China.

Department of Laboratory Medicine, Shanghai Eighth People's Hospital, Shanghai 200233, China.

出版信息

Molecules. 2016 May 11;21(5):602. doi: 10.3390/molecules21050602.

Abstract

BACKGROUND

Spermatozoa become mature and acquire fertilizing capacity during their passage through the epididymal lumen. In this study, we identified new epididymal luminal fluid proteins involved in sperm maturation in infertile rats by dutasteride, a dual 5α-reductase inhibitor, in order to provide potential epididymal targets for new contraceptives and infertility treatment.

METHODS

Male rats were treated with dutasteride for 28 consecutive days. We observed the protein expression profiles in the epididymal luminal fluids in infertile and normal rats using isobaric tags for relative and absolute quantitation (iTRAQ) technique. The confidence of proteome data was validated by enzyme-linked immunosorbent assays.

RESULTS

1045 proteins were tested, and 23 of them presented different expression profiling in the infertile and normal rats. The seven proteins were down-regulated, and 16 proteins were up-regulated. Among the seven proteins which were significantly down-regulated by dutasteride in the epididymal luminal fluids, there were three β-defensins (Defb2, Defb18 and Defb39), which maybe the key proteins involved in epididymal sperm maturation and male fertility.

CONCLUSIONS

We report for the first time that dutasteride influences the protein expression profiling in the epididymal luminal fluids of rats, and this result provides some new epididymal targets for male contraception and infertility therapy.

摘要

背景

精子在通过附睾管腔的过程中逐渐成熟并获得受精能力。在本研究中,我们通过度他雄胺(一种双重5α-还原酶抑制剂)鉴定了不育大鼠附睾管腔液中参与精子成熟的新蛋白质,以便为新型避孕药和不育症治疗提供潜在的附睾靶点。

方法

雄性大鼠连续28天接受度他雄胺治疗。我们使用相对和绝对定量的等压标签(iTRAQ)技术观察不育大鼠和正常大鼠附睾管腔液中的蛋白质表达谱。蛋白质组数据的可信度通过酶联免疫吸附测定进行验证。

结果

共检测了1045种蛋白质,其中23种在不育大鼠和正常大鼠中呈现不同的表达谱。7种蛋白质表达下调,16种蛋白质表达上调。在附睾管腔液中被度他雄胺显著下调的7种蛋白质中,有3种β-防御素(Defb2、Defb18和Defb39),它们可能是参与附睾精子成熟和男性生育能力的关键蛋白质。

结论

我们首次报道度他雄胺影响大鼠附睾管腔液中的蛋白质表达谱,这一结果为男性避孕和不育治疗提供了一些新的附睾靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0bec/6273551/05e7c22e05c7/molecules-21-00602-g001.jpg

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