Xiong C Z, Zhang L, Zhang Y, Li R W, Sun Y, Liu Y H
School of Public Health, Hubei Province Key Laboratory of Occupational Hazard Identifiction and Control, Medical College, Wuhan University of Science and Technolgy, Wuhan 430065, China.
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2023 Aug 20;41(8):562-568. doi: 10.3760/cma.j.cn121094-20220512-00258.
To investigate the role of ferroptosis in testicular injury in adolescent male mice induced by TDCIPP. In December 2021, 30 healthy 3-week-old male C57BL/6 mice, with a body weight of (13±2) g, were selected and fed adaptive for one week. They were divided into control group, low-dose group, medium-dose group, high-dose group and iron death inhibitor group according to a random number table, with 6 mice in each group. Mice in low, medium and high dose groups were treated with 5, 25 and 125 mg/ (kg·d) TDCIPP for 28 days, respectively, while the control group was treated with the same amount of corn oil for 28 days. The iron death inhibitor group was given 125 mg/ (kg·d) TDCIPP intragastric administration for 28 days, and 30 mg/kg DFO saline solution was intraperitoneally injected three times a week. After the treatment, the mice were killed, the epididymis was separated, and sperm count was performed. HE staining was used to observe the morphological changes of mouse testis, and iron content in testis was detected by tissue iron detection kit. The level of reactive cxygen species, MDA content, and the mitochondrial membrane potential level of mice were detected. Western blot analysis of testicular glutathione peroxidase (GPX4) and internal cyclooxygenase-2 (COX2) protein expression. Compared with the control group, the spermatogenic cells in the testes of mice treated with medium-and high-dose of TDCIPP were disorderly arranged, showing a vacuolar structure. the number of sperm in the epididymis was significantly reduced (=0.009, 0.004), while the sperm deformity rate was significantly increased (=0.010, 0.000). Moreover, the content of ROS, iron ion and MDA in the testes increased significantly (<0.05), and the mitochondrial membrane potential of mouse testicular cells decreased significantly (<0.05). The expression of GPX4 proteins decreased (<0.05). while the expression of COX2 increased significantly (<0.01). Compared with high-dose group group, spermatogenic cells in ferroptosis inhibitor group were closely arranged and normal, and ROS and Fe contents in testicular tissue were significantly decreased (<0.01) ; GPX4 protein expression was significantly increased while COX2 protein expression was significantly decreased (<0.05) . Ferroptosis is involved in TDCIPP-induced testicular damage in male pubertal mice.
探讨铁死亡在TDCIPP诱导的青春期雄性小鼠睾丸损伤中的作用。2021年12月,选取30只健康的3周龄雄性C57BL/6小鼠,体重为(13±2)g,进行一周适应性喂养。根据随机数字表将其分为对照组、低剂量组、中剂量组、高剂量组和铁死亡抑制剂组,每组6只。低、中、高剂量组小鼠分别用5、25和125mg/(kg·d)的TDCIPP处理28天,而对照组用等量玉米油处理28天。铁死亡抑制剂组给予125mg/(kg·d)的TDCIPP灌胃28天,并每周三次腹腔注射30mg/kg的DFO盐溶液。处理后,处死小鼠,分离附睾并进行精子计数。采用HE染色观察小鼠睾丸的形态变化,并用组织铁检测试剂盒检测睾丸中的铁含量。检测小鼠的活性氧水平、丙二醛含量和线粒体膜电位水平。通过蛋白质免疫印迹法分析睾丸中谷胱甘肽过氧化物酶(GPX4)和环氧化酶-2(COX2)的蛋白表达。与对照组相比,中、高剂量TDCIPP处理的小鼠睾丸生精细胞排列紊乱,呈空泡状结构。附睾精子数量显著减少(P=0.009,0.004),而精子畸形率显著增加(P=0.010,0.000)。此外,睾丸中活性氧、铁离子和丙二醛含量显著增加(P<0.05),小鼠睾丸细胞的线粒体膜电位显著降低(P<0.05)。GPX4蛋白表达降低(P<0.05),而COX2表达显著增加(P<0.01)。与高剂量组相比,铁死亡抑制剂组生精细胞排列紧密、正常,睾丸组织中活性氧和铁含量显著降低(P<0.01);GPX4蛋白表达显著增加,而COX2蛋白表达显著降低(P<0.05)。铁死亡参与了TDCIPP诱导雄性青春期小鼠睾丸损伤的过程。