SOX30基因敲除对雄性小鼠睾丸组织结构及激素分泌的影响

doi:10.3969/j.issn.1004-616x.2020.05.001

癌变·畸变·突变 ›› 2020, Vol. 32 ›› Issue (5): 329-335.doi: 10.3969/j.issn.1004-616x.2020.05.001

• 论著 • 下一篇

SOX30基因敲除对雄性小鼠睾丸组织结构及激素分泌的影响

王平^1,2, 姜晓¹, 韩飞¹, 崔志鸿³, 唐莹^1,4, 周洋西^1,3, 刘文斌¹, 曹佳¹, 刘晋祎¹

1. 陆军军医大学军事预防医学系毒理学研究所, 重庆 400038;
2. 陆军军医大学第二附属医院血液病医学中心, 重庆 400037;
3. 西南大学药学院, 重庆 400715;
4. 宁夏医科大学公共卫生与管理学院劳动卫生与环境卫生学教研室, 宁夏银川 750004

收稿日期:2020-07-22 修回日期:2020-08-22 出版日期:2020-10-01 发布日期:2020-10-12
通讯作者: 刘晋祎,E-mail:jinyiliutmmu@163.com E-mail:jinyiliutmmu@163.com
作者简介:王平,E-mail:wpii_2002@sina.com。
基金资助:
重庆市基础科学与前沿技术研究重点项目（cstc2017jcyjBX 0064）；国家自然科学基金（81773461）

Effects of SOX30 gene knockout on testicular tissue structure and hormone secretion in male mice

WANG Ping^1,2, JIANG Xiao¹, HAN Fei¹, CUI Zhihong³, TANG Ying^1,4, ZHOU Yangxi^1,3, LIU Wenbin¹, CAO Jia¹, LIU Jinyi¹

1. Institute of Toxicology, College of Preventive Medicine, Army Medical University, Chongqing 400038;
2. Medical Center of Hematology, The Xinqiao Hospital of Army Medical University, Chongqing 400037;
3. College of Pharmaceutical Sciences and Chinese Medicine, Southwest University, Chongqing 400715;
4. Department of Occupational Health and Environmental Hygiene, School of Public Health and Management, Ningxia Medical University, Yinchuan 750004, Ningxia, China

Received:2020-07-22 Revised:2020-08-22 Online:2020-10-01 Published:2020-10-12

摘要/Abstract

摘要： 目的：分析SOX30基因敲除后对雄性小鼠睾丸组织结构及激素分泌的影响，探讨SOX30基因在雄性生殖系统中的作用。方法：采用基因重组方法构建SOX30基因敲除小鼠模型，并根据基因型鉴定结果将小鼠分为野生型（WW）、杂合型（KW）和纯合型（KK）共3组，分别称取各基因型小鼠（2.5月龄）体质量和各脏器质量并计算脏器系数。HE染色后观察各基因型小鼠睾丸及附睾病理改变并测量统计睾丸曲细精管直径。酶联免疫吸附法（ELISA）检测睾丸组织匀浆液中睾酮（T）和抑制素B（INH-B）激素水平。免疫荧光法检测睾丸组织中支持细胞和间质细胞特异性蛋白SOX9和3β-HSD的表达变化。结果：与WW型和KW型比较，KK型小鼠睾丸脏器指数显著降低（P < 0.05），曲细精管内生精细胞排列紊乱，可见巨细胞及空泡变性，精子数量显著减少（P < 0.05），曲细精管外间质增生明显，附睾内未发现成熟精子。与WW型比较，KK型小鼠曲细精管管腔直径缩小（P < 0.05）。与WW型和KW型比较，KK型小鼠睾丸组织匀浆液中T和INH-B水平显著降低（P < 0.05）。WW型和KK型小鼠睾丸组织中SOX9蛋白表达阳性细胞数量无明显差异，而KK型小鼠睾丸组织中3β-HSD蛋白表达阳性细胞数量较WW型明显升高（P < 0.05）。结论：SOX30敲除可导致睾丸组织出现明显的病理损伤，SOX30参与调控小鼠睾丸T和INH-B激素合成，在维持睾丸的正常功能方面具有重要作用。

关键词: SOX30基因, 基因敲除, 雄性, 小鼠, 生殖

Abstract: OBJECTIVE: To evaluate the effects of SOX30 gene knockout on testicular tissue structure and hormone secretion in male mice,and to explore the role of SOX30 gene in male reproductive system. METHODS: The SOX30 gene knockout mouse model was constructed by the gene recombination method and genotypes were identified by the PCR method. The male mice were divided into wild type (WW),heterozygous (KW) and homozygous (KK) groups according to the results from genotype identification (2.5 months old). Testicular volume and testicular organ index were calculated. After H&E staining,structures of the testes and epididymes of each mouse were determined,and diameters of the Seminiferous tubule were measured. Levels of the testosteron (T) and inhibin B (INH-B) hormones in homogenates of testicular tissues were detected by ELISA. Expressions of SOX9 and 3β-HSD in testicular tissues were determined by using the immunofluorescence technique. RESULTS: Compared with the WW and KW types,the testicular organ index of KK mice was significantly decreased (P < 0.05). There were no pathological changes in the testes and epididymes of KW and WW mice. On the contrary,testes in KK mice had disordered spermatogenic cells,giant cells,vacuolar degeneration and interstitial hyperplasia. No matured sperm was found in the epididymis of KK mice. The lumen diameters of seminiferous tubules in the KK mice were reduced compared with the WW mice (P < 0.05). In addition,the T and INH-B levels in homogenates from testicular tissues were reduced significantly in the KK compared with the WW and KW mice (P < 0.05). There was no significant difference in the number of Sertoli cells expressing SOX9 between WW and KK mice,but the number of leydig cells expressing 3β-HSD in KK mice was significantly more than that in WW mice (P < 0.05). CONCLUSION: Our results suggest that the knockout of SOX30 induced pathological damage in testes of mice via inhibiting the synthesis of T and INH-B which played important roles in maintaining normal functions of testes.

Key words: SOX30 gene, gene knockout, male, mice, reproduction

中图分类号:

Q492.4

王平, 姜晓, 韩飞, 崔志鸿, 唐莹, 周洋西, 刘文斌, 曹佳, 刘晋祎. SOX30基因敲除对雄性小鼠睾丸组织结构及激素分泌的影响[J]. 癌变·畸变·突变, 2020, 32(5): 329-335.

WANG Ping, JIANG Xiao, HAN Fei, CUI Zhihong, TANG Ying, ZHOU Yangxi, LIU Wenbin, CAO Jia, LIU Jinyi. Effects of SOX30 gene knockout on testicular tissue structure and hormone secretion in male mice[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2020, 32(5): 329-335.

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SOX30基因敲除对雄性小鼠睾丸组织结构及激素分泌的影响

Effects of SOX30 gene knockout on testicular tissue structure and hormone secretion in male mice

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