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目的通过17-β雌二醇诱导小鼠隐睾症模型,检测Lgr8在小鼠睾丸中的表达,探讨其在雌二醇所致隐睾症中的可能作用。方法将47只新生BALB/c雄性小鼠随机分为3个组,即正常对照组(15例)、溶剂对照组(16例)及实验组(16例)。在3 d龄至35 d龄期间,实验组皮下注射4μg/d 17-β雌二醇,溶剂对照组注射4μg/d的丙二醇,正常对照组不给予任何处理。第35天处死各组小鼠。观察隐睾发生率,测量睾丸重量、附睾重量及脏器系数,对仔鼠睾丸进行组织病理学观察,测量精曲小管直径及面积。免疫组织化学、RT-q PCR分别检测仔鼠睾丸中的Lgr8蛋白和Lgr8mRNA。结果正常对照组和丙二醇组仔鼠隐睾发生率均为0;雌二醇组仔鼠发生隐睾率100%。与正常对照组及丙二醇组相比,雌二醇组的睾丸重量、附睾重量和脏器系数降低(P<0.05),雌二醇组每日精子生成量、精子活动率及附睾悬液精子计数和降低(P<0.05),而精子畸形率明显升高(P<0.01)。Lgr8蛋白在正常对照组和丙二醇组的表达明显高于雌二醇组(P<0.05),但雌二醇组Lgr8 mRNA的相对表达量明显高于对照组和丙二醇组(P<0.05)。结论雌二醇可诱导BALB/c雄性小鼠发生隐睾症,有可能通过影响Lgr8转录后水平介导隐睾症的发生。
Objective To explore the possible role of Lgr8 in the testis of mice induced by estradiol by inducing mouse model of cryptorchidism by 17-β estradiol. Methods 47 neonatal BALB / c male mice were randomly divided into three groups: normal control group (15 cases), solvent control group (16 cases) and experimental group (16 cases). During 3 days to 35 days of age, 4μg / d 17-β-estradiol was injected subcutaneously in the experimental group and 4μg / d of propylene glycol was injected in the solvent control group. No treatment was given in the normal control group. The mice were sacrificed on the 35th day. The incidence of cryptorchidism was observed. The testicular weight, epididymal weight and organ coefficient were measured. The testes of the offspring were observed histopathologically, and the diameter and area of the tubule were measured. Immunohistochemistry and RT-q PCR were used to detect Lgr8 protein and Lgr8 mRNA in testis of offspring respectively. Results The incidence of cryptorchidism in the normal control group and the propylene glycol group were all 0; the rate of cryptorchidism in the estradiol group was 100%. Compared with the normal control group and the propylene glycol group, the testis weight, epididymis weight and organ coefficient decreased (P <0.05) in the estradiol group. The daily sperm production, sperm motility and the sperm count of the epididymis suspension And decreased (P <0.05), while the sperm deformity rate was significantly increased (P <0.01). The expression of Lgr8 protein in normal control group and propylene glycol group was significantly higher than that in estradiol group (P <0.05), but the relative expression of Lgr8 mRNA in estradiol group was significantly higher than that in control group and propylene glycol group (P <0.05). Conclusion Estradiol can induce cryptorchidism in BALB / c male mice, which may affect the development of cryptorchidism by affecting the post-transcriptional level of Lgr8.