白消安对小鼠睾丸损伤机理的研究

旨在揭示白消安处理对睾丸的损伤机理，提高精原干细胞（SSCs）移植受体制备效率及安全性，缓解或避免因白消安毒性导致的雄性不育。对白消安处理小鼠进行睾丸生物素示踪试验，以验证血睾屏障（BTB）完整性；采用液质联用靶向测定其附睾尾精液中白消安的含量，以验证白消安是否可以直接穿过睾丸BTB进入曲细精管；并对小鼠血清进行炎性因子ELISA测定，以验证其浓度是否发生变化。结果发现，小鼠睾丸注射白消安后，曲细精管内最早24 h可检测到生物素示踪红色荧光，表明BTB已被破坏；注射白消安30 min后，附睾精液中检测到最高水平白消安，随后，其水平逐渐降低；小鼠睾丸注射白消安后，血清中TNF-α、IL-1β和IL-6浓度逐渐增加，36 h后均显著高于对照组（P<0.05），21 d后达到峰值时，TNF-α浓度为（1 855.51±10.32） pg·mL^-1，IL-1β浓度为（293.59±3.34） pg·mL^-1，IL-6浓度为（340.30±12.55） pg·mL^-1，随后逐渐下降。综上表明，白消安可自由穿越BTB，且于24 h生物示踪素可突破BTB，血清炎性因子TNF-α、IL-1β和IL-6浓度逐渐升高并达显著水平，在21 d达到最高后逐渐下降。

Study on the Damage Mechanism of Busulfan on Mouse Testis

The study aimed to improve the efficiency and safety of spermatogonial stem cells (SSCs) transplantation recipients so that to decrease male sterility caused by busulfan toxicity, which is helpful in revealing the damage mechanism of busulfan on testis. Testicular biotin tracing experiments were performed on busulfan-treated mice to verify the integrity of the blood testis barrier (BTB); Liquid-mass spectrometry was used to target the content of busulfan in the semen of the epididymal tail to verify whether busulfan could directly pass through the testis BTB into the seminiferous tubules; ELISA determination of inflammatory factors was performed in mouse serum to verify whether its concentration had changed. The results showed that the earliest observable red fluorescence in the seminiferous tubules traced by biotin was detected at 24 h, suggesting the damage of blood testis barrier (BTB). And the highest proportion of busulfan in the epididymal semen reached a summit at 30 min post-treatment then gradually decreased. In serum, the concentration of TNF-α, IL-1β and IL-6 progressively increased by busulfan treatment(P<0.05), and at 36 h the amount of the three indexes was significantly higher than that in the control group. The values peaked at 21 d with TNF-α of (1 855.51±10.32) pg·mL^-1, IL-1β of (293.59±3.34) pg·mL^-1,and IL-6 of (340.30±12.55) pg·mL^-1, then gradually decreased. In summary, busulfan could freely cross over BTB and the biotracer could break through BTB within 24 h. The serum inflammatory factors TNF-α, IL-1β and IL-6 concentration gradually increased and reached significant levels, reaching the highest level at 21 d, then gradually decreased.