BALB/c鼠自身免疫性睾丸炎动物模型的制作

Animal Model of Experimental Autoimmunity Orchitis in BALB/c Mice

【目的】探讨BALB/c鼠睾丸炎动物模型的建立方法,观察睾丸炎形成后到生殖功能恢复的时间间隔,以及该动物模型的稳定性。【方法】BALB/c小鼠(10周)背脊皮下注射睾丸混悬液0.1mL,浓度2×107/mL,1次/周,共4次。同种睾丸混悬液(组1)、同种睾丸混悬液加完全免疫佐剂(组2)、异种睾丸混悬液(组3)、异种睾丸混悬液加完全免疫佐剂(组4)、空白对照(组5)。【结果】组3建模成功率80%,第5、8、11周按Johusen评分为5.2±1.44、6.2±0.79、8.1±0.87,组1、2、5评分均在9.0以上,组3生精功能明显降低(P<0.05)。组4小鼠于第2次注射后3～5d,全部死亡。组3第5周生精上皮细胞减少至2￣3层,曲细精管管腔中有稀疏的精子,间质出现水肿,淋巴细胞浸润约10%￣25%。第5周组3附睾尾精子浓度为(11.0±6.2)×106,精子存活率(10.73±8.14)%,精子活动率(7.3±6.1)%,明显低于组1、2、5(P<0.05),于第11周组3恢复至正常水平。睾丸炎生精障碍持续21￣28d。【结论】异种小鼠生殖细胞可诱导建立BALB/c小鼠的EAO模型,模型有一定的持续稳定性,可用于精子发育生物学研究。

[Objective] To establish the experimental autoimmunity orchitis （EAO） in BALB/c and to observe the interval of hypospermatogenesis and the stability of EAO. [Method] Mate BALB/c mice （10 weeks） were injected subcutaneously once a week with 2×10^7testicular germ cells in 0.1 mL of Hank＇s balanced saline solution （HBSS） over 4 weeks. Group 1 was treated with syngeneic testicular germ cells, group 2 with syngeneic germ cell and adjuvant, group 3 with heterogeneity germ cell, group 4 with heterogeneity germ cell and adjuvant. Group 5 was control group.[Result] The success rate for model establishment of the EAO in group 3 was 80%. Spermatogenesis was graded by Johusen score in group 3 and the scores were lower than the other groups,which were 5.2±1.44, 6.2±0.79, and 8.1±0.87, respectively, in 5th week, 8th week, and 11th week after the frrst injection （P〈 0.05）. The scores of group 1, group 2, and group 5 were more than 9,0. All mice in group 4 were dead within 3-5 days after the 2nd injection. The germinal epithelium in group 3 was reduced to 2-3 layers, sperms were sparse in the seminfferous tubules. Edema appeared in interstitiat and lymphocytes were infiltrated 10%-25% or more of the whole area of the section. Number of epididymat sperm in group 3 in 5th week was （11.0±6.2）×10^6, which was significant less than that in the other groups （P〈 0.05）. Besides, the sperm viability （10.73%±8.14%） and motility （7.3%＋6.1%） were lower than the other groups （P〈 0.05）. The sperm number in group 3 restored to normat level in 11th week. There was not significant difference in group 1, 2, and 5. The state of hypospermatogenesis in EAO can last 21-28 days. [Conclusion] The heterogeneity germ cells can induce EAO in male BALB/c. It can facilitate the study of the regulation of spermatogenesis during the stable period of EAO. It suggested the EAO should content 2 points： Histopathology in Johusen score should be less than 8 or the immature germ cells were more than 50% in seminiferous tubules. The viability and motility in epididymis were lower than 25% and 15%. Lymphocytes infiltration was over 20% in the section.