Li Hui-Yun, Song Chun-Ying, Zhao Jun, Wang Huai-Xiu, Li Hong-Jun, Guo Xing-Ping
Faculty of Biochemistry and Molecular Biology, Shanxi Medical University, Taiyuan, Shanxi 030001, China.
Key Laboratory of Shanxi Province for Birth Defects and Cell Regeneration, Shanxi Research Institute of Reproductive Science, Taiyuan, Shanxi 030006, China.
Zhonghua Nan Ke Xue. 2019 Aug;25(8):681-689.
To explore the pathways of development and maturation of the testis tissue in mice with spermatogenic dysfunction in vitro.
Sixty-eight 8-week-old BALB/c male mice were randomly divided into four groups of equal number, normal control, Sertoli cell only syndrome (SCOS), severe hypo-spermatogenesis (H-S1), and mild hypo-spermatogenesis (H-S2), and the models were established in the latter three groups by intraperitoneal injection of busulfan at 40 mg/kg for 4, 6 and 8 weeks, respectively. The testis tissues of the mice were cultured with the agarose gel method in vitro till the 4th week, followed by determination of the expressions of the marker proteins STRA8 at meiotic initiation, SCP3 during meiosis, and TNP1 after meiosis by immunohistochemistry. The development and maturation of the germ cells during the in vitro culture were evaluated, and their apoptosis detected by TUNEL.
The more severe the testicular tissue injury, the lower the expression of the STRA8 protein in the SCOS, H-S1 and H-S2 groups as compared with the normal control before in vitro culture on agarose gel (P < 0.05), and the STRA8 expression was significantly upregulated in the former three groups after 4 weeks of culturing (P < 0.05). The expression of SCP3 was the lowest in the SCOS but the highest in the H-S2 group before culturing (P < 0.05), and was not as high as that in the control, though increased after 4 weeks of culturing. TNP1 was positively expressed in all the mice of the control, some individuals of the H-S1 and H-S2 groups (P< 0.05), but not in the SCOS group at 4 weeks. The apoptosis of germ cells was significantly increased in the SCOS but decreased in the H-S groups compared with that in the normal control after 4 weeks of culturing (P< 0.05).
In vitro culture on agarose gel induces the meiosis of the testis tissue in BALB/c mice with spermatogenic dysfunction, and the effect is even better in those with mild spermatogenic dysfunction.
探讨体外培养生精功能障碍小鼠睾丸组织的发育及成熟途径。
将68只8周龄BALB/c雄性小鼠随机分为4组，每组数量相等，即正常对照组、唯支持细胞综合征（SCOS）组、重度少精子症（H-S1）组和轻度少精子症（H-S2）组。后三组分别腹腔注射40mg/kg白消安4周、6周和8周建立模型。采用琼脂糖凝胶法体外培养小鼠睾丸组织至第4周，然后通过免疫组织化学法检测减数分裂起始标记蛋白STRA8、减数分裂期间的SCP3以及减数分裂后的TNP1的表达。评估体外培养过程中生殖细胞的发育和成熟情况，并通过TUNEL法检测其凋亡情况。
与体外培养于琼脂糖凝胶前的正常对照组相比，SCOS组、H-S1组和H-S2组睾丸组织损伤越严重，STRA8蛋白表达越低（P<0.05）；培养4周后，前三组STRA8表达显著上调（P<0.05）。培养前SCP3表达在SCOS组最低，在H-S2组最高（P<0.05），培养4周后虽有所升高，但仍不及对照组。4周时，TNP1在对照组所有小鼠、H-S1组和H-S2组部分个体中呈阳性表达（P<0.05），而SCOS组未见表达。培养4周后，与正常对照组相比，SCOS组生殖细胞凋亡显著增加，H-S组则减少（P<0.05）。
琼脂糖凝胶体外培养可诱导生精功能障碍的BALB/c小鼠睾丸组织减数分裂，对轻度生精功能障碍小鼠的效果更佳。
