Dai Yanlin, Sun Lihui, Han Shanshan, Xu Shanshan, Wang Long, Ding Ying
Pediatric Medical College, Henan University of Traditional Chinese Medicine, Zhengzhou, China.
Department of Pediatrics, The First Affiliated Hospital of Henan University of Traditional Chinese Medicine, Zhengzhou, China.
Front Pharmacol. 2022 May 20;13:888968. doi: 10.3389/fphar.2022.888968. eCollection 2022.
glycoside tablet (TGT) is a common clinically used and effective non-steroidal immunosuppressant. However, its reproductive toxicity limits its application in pediatric immune diseases, warranting the study of the molecular mechanism behind its reproductive toxicity. In the present study, 4-week-old male Sprague Dawley (SD) rats were provided TGT through continuous gavage with a clinically equivalent dose of 12 mg/kg for 12 weeks. The reproductive toxicity of TGT was recorded, and its toxicity mechanism was verified through experimental validation and proteomics analyses. Our results demonstrated that TGT could significantly reduce the testosterone level in the serum as well as the concentration and survival rate of sperms. Pathological sections of the testis revealed that TGT could reduce spermatocytes at different levels and make the convoluted meridians vacuolated. Based on tandem mass tag (TMT)-labeled quantitative rats testicular tissue proteomics, 34 differential proteins were screened, involving protein digestion and absorption, PPAR signaling pathway, PI3K-Akt, and other pathways, among which PI3K-Akt plays an important role in the study of reproductive injury. Western blotting results revealed that TGT could significantly downregulate the Col1A1, Col1A2, p-PI3K, and p-Akt expressions and inhibit the expression of proteins related to the PI3K-Akt signaling pathway. In summary, the clinically equivalent dose of TGT induced reproductive toxicity of 4-week-old male SD rats, possibly in relation to the inhibition of the PI3K-Akt pathway expression.
糖甙片（TGT）是临床常用且有效的非甾体类免疫抑制剂。然而，其生殖毒性限制了它在儿童免疫疾病中的应用，因此有必要研究其生殖毒性背后的分子机制。在本研究中，对4周龄雄性Sprague Dawley（SD）大鼠连续灌胃给予临床等效剂量12 mg/kg的TGT，持续12周。记录TGT的生殖毒性，并通过实验验证和蛋白质组学分析验证其毒性机制。我们的结果表明，TGT可显著降低血清睾酮水平以及精子浓度和存活率。睾丸病理切片显示，TGT可不同程度减少精母细胞数量，并使曲细精管出现空泡化。基于串联质谱标签（TMT）标记的定量大鼠睾丸组织蛋白质组学，筛选出34种差异蛋白，涉及蛋白质消化吸收、PPAR信号通路、PI3K-Akt等通路，其中PI3K-Akt在生殖损伤研究中起重要作用。蛋白质免疫印迹结果显示，TGT可显著下调Col1A1、Col1A2、p-PI3K和p-Akt的表达，并抑制与PI3K-Akt信号通路相关的蛋白质表达。综上所述，临床等效剂量的TGT可诱导4周龄雄性SD大鼠产生生殖毒性，可能与抑制PI3K-Akt通路表达有关。
