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目的:探讨精子再生过程中Ki-67、Thy-1和Oct4的表达及其意义。方法:采用间隔24d2次白消安[10mg/(kg.次)]小鼠腹腔内注射建立精子再生模型,于第2次给药后第1,2,3,4,6和8周运用免疫组化检测Ki-67、Thy-1和Oct4在生精上皮表达及其表达规律。结果:形态学显示第2次给药后3周主要为精原干细胞增殖期,4周为分化期,6-8周为精子发生恢复期。Ki-67表达见于精原细胞和少量精母细胞。精原细胞Ki-67阳性率在第2次给药后3周最高,第4周明显下降,6-8周恢复至正常对照组水平。Thy-1表达于精原细胞,在精子再生各阶段阳性表达率均较高,其中,第3周表达最高,第2周次之。Oct4表达仅见于紧贴曲细精管基底膜的精原细胞,在第3周表达较对照组显著上调,第4周下降,6-8周恢复至对照组水平。精原细胞Oct4和Ki-67阳性率之间的相关系数为0.779。结论:Thy-1并非特异性表达于精原干细胞。Oct4表达上调是促进精原干细胞增殖的重要因素。
Objective: To investigate the expression and significance of Ki-67, Thy-1 and Oct4 during spermatogenesis. METHODS: A rat model of spermatogenesis was established by intraperitoneal injection of busulfan [10 mg / (kg · s)] at intervals of 24 days. The mice were immunized at 1, 2, 3, 4, 6 and 8 weeks after the second administration Expressions of Ki-67, Thy-1 and Oct4 in the seminiferous epithelium and their expressions were detected by histochemical staining. RESULTS: Morphology showed that the spermatogonial stem cells were mainly proliferated at 3 weeks after the second administration, with a differentiation period of 4 weeks and a recovery period of 6 to 8 weeks. Ki-67 expression seen in spermatogonia and a small amount of spermatocytes. The positive rate of Ki-67 in spermatogonia was the highest at 3 weeks after the second administration, decreased significantly at the 4th week and recovered to the normal control group at 6-8 weeks. Thy-1 was expressed in spermatogonia, and the positive expression rate of Thy-1 was higher in all stages of spermatogenesis. Among them, the highest expression was found in the third week and the second week was followed. Oct4 expression was seen only in spermatogonia clinging to the basement membrane of the seminiferous tubules. The expression of Oct4 was significantly up-regulated in the third week compared with the control group, decreased in the fourth week and returned to the control group in 6-8 weeks. The correlation coefficient between the positive rates of Oct4 and Ki-67 in spermatogonia was 0.779. Conclusion: Thy-1 is not specifically expressed in spermatogonial stem cells. Oct4 upregulation is an important factor to promote the proliferation of spermatogonial stem cells.