Establishing a nonlethal and efficient mouse model of male gonadotoxicity by intraperitoneal busulfan injection
Establishing a nonlethal and efficient mouse model of male gonadotoxicity by intraperitoneal busulfan injection
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腹腔注射白消安建立非致死高效雄性性腺毒性小鼠模型
DOI：
10.4103/aja.aja_41_19
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发表时间：
2020-03-01
影响因子：
2.9
通讯作者：
Liu, Gui-Hua
中科院分区：
文献类型：
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作者：
Xie, Yun;Deng, Cun-Can;Liu, Gui-Hua
理想的无精子症动物模型是评价精原干细胞移植效果的有力工具。白消安已被普遍用于开发这样的模型，但30%-87%的小鼠死亡时，给予腹腔注射40毫克kg-1。在本研究中，使用苏木精和伊红染色，Western印迹，免疫荧光和定量实时聚合酶链反应来测试白消安暴露在小鼠模型中的影响，该模型接受两次不同剂量的白消安腹腔注射，间隔3小时（20、30和40 mg kg-1）或在不同时间点（0、9、18、27、36和63天）给予40 mg kg-1剂量。小鼠的存活率为100%。当小鼠用40 mg kg-1白消安处理时，18天后发生了显著的SSC耗竭，并且所有生殖细胞在第36天被清除。此外，胶质细胞源性神经营养因子（GDNF）、成纤维细胞生长因子2（FGF 2）、趋化因子（C-X-C基序）配体12（CXCL 12）和集落刺激因子1（CSF 1）的基因表达在第36天适度增加。63天的长期观察显示，睾丸中内源性生殖细胞的恢复很少，这表明SSC移植的潜在时期在第36天和第63天之间。我们的研究结果表明，白消安（40毫克kg-1的总）在3小时的间隔，小鼠腹腔注射给药提供了一个非致命的和有效的方法，在SSC移植受体的准备，可以提高不孕症的治疗和化疗引起的性腺毒性的理解。
An ideal animal model of azoospermia would be a powerful tool for the evaluation of spermatogonial stem cell (SSC) transplantation. Busulfan has been commonly used to develop such a model, but 30%-87% of mice die when administered an intraperitoneal injection of 40 mg kg-1. In the present study, hematoxylin and eosin staining, Western blot, immunofluorescence, and quantitative real-time polymerase chain reaction were used to test the effects of busulfan exposure in a mouse model that received two intraperitoneal injections of busulfan at a 3-h interval at different doses (20, 30, and 40 mg kg-1) on day 36 or a dose of 40 mg kg-1 at different time points (0, 9, 18, 27, 36, and 63 days). The survival rate of the mice was 100%. When the mice were treated with 40 mg kg-1 busulfan, dramatic SSC depletion occurred 18 days later and all of the germ cells were cleared by day 36. In addition, the gene expressions of glial cell line-derived neurotrophic factor (GDNF), fibroblast growth factor 2 (FGF2), chemokine (C-X-C Motif) ligand 12 (CXCL12), and colony-stimulating factor 1 (CSF1) were moderately increased by day 36. A 63-day, long-term observation showed the rare restoration of endogenous germ cells in the testes, suggesting that the potential period for SSC transplantation was between day 36 and day 63. Our results demonstrate that the administration of two intraperitoneal injections of busulfan (40 mg kg-1 in total) at a 3-h interval to mice provided a nonlethal and efficient method for recipient preparation in SSC transplantation and could improve treatments for infertility and the understanding of chemotherapy-induced gonadotoxicity.