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The mechanism of steroidogenesis and spermatogenesis impairment in men with type 2 diabetes remains unclear. We aimed to explore the local changes of steroidogenesis and spermatogenesis in the testis of db\/db mice. <jats:italic>Research Design and Methods<\/jats:italic>. We performed single-cell RNA sequencing analysis in the testis of db\/db and C57BL\/6J mice. The differentially expressed genes were then confirmed by real-time PCR. The histopathological characteristics of testis in db\/db mice and C57BL\/6J control were also performed. <jats:italic>Results<\/jats:italic>. The 20-week-old db\/db mice had significantly higher blood glucose and body weight (both <mml:math xmlns:mml=\"http:\/\/www.w3.org\/1998\/Math\/MathML\" id=\"M1\"><mml:mi>p<\/mml:mi><mml:mo>&lt;<\/mml:mo><mml:mn>0.001<\/mml:mn><\/mml:math>). The serum testosterone levels (4.4\u2009\u00b1\u20090.8 vs. 9.8\u2009\u00b1\u20090.7\u2009ng\/ml, <mml:math xmlns:mml=\"http:\/\/www.w3.org\/1998\/Math\/MathML\" id=\"M2\"><mml:mi>p<\/mml:mi><mml:mo>=<\/mml:mo><mml:mn>0.001<\/mml:mn><\/mml:math>) and weight of the testis (0.16\u2009\u00b1\u20090.01 vs. 0.24\u2009\u00b1\u20090.01\u2009g, <mml:math xmlns:mml=\"http:\/\/www.w3.org\/1998\/Math\/MathML\" id=\"M3\"><mml:mi>p<\/mml:mi><mml:mo>&lt;<\/mml:mo><mml:mn>0.001<\/mml:mn><\/mml:math>) were significantly lower in db\/db mice than that in C57BL\/6J controls. db\/db mice had a lower cross-sectional area of seminiferous tubules and thickness of the cell layer (both <mml:math xmlns:mml=\"http:\/\/www.w3.org\/1998\/Math\/MathML\" id=\"M4\"><mml:mi>p<\/mml:mi><mml:mo>&lt;<\/mml:mo><mml:mn>0.05<\/mml:mn><\/mml:math>). The numbers of Sertoli cells and Leydig cells decreased in db\/db mice (both <mml:math xmlns:mml=\"http:\/\/www.w3.org\/1998\/Math\/MathML\" id=\"M5\"><mml:mi>p<\/mml:mi><mml:mo>&lt;<\/mml:mo><mml:mn>0.01<\/mml:mn><\/mml:math>). Single-cell RNA sequencing analysis showed that compared with the control group, the percentage of spermatogonia was significantly higher in the db\/db mouse (<mml:math xmlns:mml=\"http:\/\/www.w3.org\/1998\/Math\/MathML\" id=\"M6\"><mml:mi>p<\/mml:mi><mml:mo>&lt;<\/mml:mo><mml:mn>0.001<\/mml:mn><\/mml:math>), while the proportions of spermatocytes, round and elongating spermatids, and sperms were all lower in the db\/db mouse (<mml:math xmlns:mml=\"http:\/\/www.w3.org\/1998\/Math\/MathML\" id=\"M7\"><mml:mi>p<\/mml:mi><mml:mtext>\u2009all<\/mml:mtext><mml:mo>&lt;<\/mml:mo><mml:mn>0.001<\/mml:mn><\/mml:math>). The most differentially expressed genes were found in round spermatids (<jats:italic>n<\/jats:italic>\u2009=\u200986), which were not found in spermatogonia, spermatocyte, and sperm. Igfbp5 was the most significantly decreased gene in Leydig cells of the db\/db mouse, while the expression of Cd74, H2-Aa, and H2-Eb1 was elevated. Ccl7 and Ptgds were the most significantly increased and decreased genes in Sertoli cells of the db\/db mouse. <jats:italic>Conclusions<\/jats:italic>. The present study indicates spermiogenesis and steroidogenesis defects in db\/db mice. The mechanism of steroidogenesis impairment in the testis of db\/db mice deserves further investigation.<\/jats:p>","DOI":"10.1155\/2024\/8797972","type":"journal-article","created":{"date-parts":[[2024,5,23]],"date-time":"2024-05-23T20:35:09Z","timestamp":1716496509000},"page":"1-11","source":"Crossref","is-referenced-by-count":7,"title":["Single-Cell RNA Sequencing Analysis of Steroidogenesis and Spermatogenesis Impairment in the Testis of db\/db Mice"],"prefix":"10.1155","volume":"2024","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4477-3641","authenticated-orcid":true,"given":"Yun","family":"Hu","sequence":"first","affiliation":[{"name":"Department of Endocrinology, The Affiliated Wuxi People\u2019s Hospital of Nanjing Medical University, Wuxi People\u2019s Hospital, Wuxi Medical Center, Nanjing Medical University, Wuxi, China"},{"name":"Department of Endocrinology, Nanjing First Hospital, Nanjing Medical University, Nanjing, Jiangsu, 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