睾丸特异表达基因-1在乙醇致小鼠睾丸损伤模型中的表达及意义

Expression of testis specific expressed gene 1 in ethanol-Induced mouse testis injury model and significance

目的 探讨睾丸特异表达基因-1（TSEG-1）在乙醇致小鼠睾丸损伤模型中的表达变化及意义。方法 首先建立乙醇致睾丸损伤模型,将12只雄性昆明小鼠随机分为对照组和实验组,每组6只,用生理盐水稀释无水乙醇为15%浓度。实验组小鼠每天按3 g乙醇（15%,V/V）/kg体质量,腹腔内注射14 d,对照组为等体积生理盐水。应用苏木素-伊红（HE）染色鉴定睾丸组织形态学变化,实时定量逆转录聚合酶链反应（RT-qPCR）检测TSEG-1 mRNA变化,采用免疫组织化学和Western blot检测其蛋白变化。结果 乙醇组睾丸HE染色结果提示,与对照组比较,乙醇组睾丸组织85.6%曲精小管管腔消失,I-Ⅳ级生精细胞数目减少,78.2%精母细胞或精子细胞核皱缩。在乙醇组睾丸组织中,TSEG-1 mRNA表达水平（7.500±0.657,n=6）较对照组（0.985±0.231,n=6）显著增加,差异有统计学意义（t=22.915,P〈0.01）。同时,乙醇组TSEG-1蛋白质较生理盐水组显著增加,经Quantity One软件分析灰度值后,与管家基因甘油醛-3-磷酸脱氢酶（GAPDH）比较,乙醇组TSEG-1蛋白（1.360±0.202,n=6）较生理盐水组（0.330±0.112,n=6）表达增加达4倍,差异有统计学意义（t=10.69,P〈0.01）。结论 乙醇致小鼠睾丸损伤模型是研究睾丸损伤的可靠模型,TSEG-1可能参与乙醇致睾丸损伤模型的分子发生过程。

Objective To explore the cellular localization and transcript levels of testis specific expressed gene 1 （ TSEG-1 ） in ethanol-induced mouse testis injury model. Methods Twelve male Kunruing mice were divided into normal saline solution group （ n = 6） and ethanol injection group （ n = 6）. The morphological changes of testes were observed by hematoxylin and eosin （HE） staining. The transcriptional and protein levels of TSEG-1 were detected by real-time reverse transcriptase-polymerase chain reaction （RT-qPCR）, Immunohistochemistry （IHC） and Western blotting. Results Intraperitoneal injection of 3 g ethanol （15% , V/V）/kg body weight for 14 days induced mouse testis injury. As compared with normal saline solution group, 85.6% serminous tubules disappeared, the number of I-IV-stage spermatogenic cells was reduced, and 78.2% spermatocytes and spermatids had nuclei shrinkage in ethanol injection group. Extensive TSEG-1 mRNA levels were observed in control testes, mainly localizing in spermatocytes and early spermatids. The transcripts of TSEG-1 （7. 500 ±0. 657 ,n = 6） were up-regulated to 8 folds in ethanol injection group as compared with those in normal saline solution control group （0. 985 ±0. 231 ,n = 6）, with the difference being significant （t = 22. 915,P〈 0.01 ）. Especially, the protein expression of TSEG-1 （ 1. 360 ±0. 202, n = 6） was up-regulated to 4 folds in ethanol injection group as compared with that in normal saline solution control group （0. 330± 0. 112, n = 6）, with the difference being significant, t = 10. 69,P 〈 0. 01. Conclusion Ethanol-induced testis injury model was successfully constructed by intraperitoneal injection of ethanol. The novel gene TSEG-1 may play a role in the pathogenesis of ethanol-induced testis iniurv model, which laid the basis for further study on its function in testis diseases.