目的 通过检测增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)及Ki67在正常及糖尿病小鼠睾丸组织中表达的差异,探讨糖尿病对生精细胞增殖的影响.方法 30只正常雄性C57BL/6小鼠,随机分为对照组(10只)和糖尿病组(20只).采用腹腔注射链脲佐菌素的方法建立糖尿病小鼠模型,对照组注射相同体积的柠檬酸—柠檬酸钠缓冲液.造模成功3周后处死动物,取睾丸组织,常规固定、石蜡包埋、切片HE染色观察睾丸组织的形态学变化;免疫组织化学SP法检测PCNA及Ki67在睾丸组织中的表达;图像分析技术分析组织中PCNA及Ki67免疫组织化学表达水平.结果 HE染色显示,糖尿病小鼠睾丸内处于精子发生前半期的生精小管内生精细胞排列疏松,细胞层数偏少,附睾管内精子密度相对较低;免疫组织化学染色显示,PCNA和Ki67在糖尿病小鼠睾丸生精小管中的表达均明显降低.结论 高糖可能通过降低生精细胞的增殖进而影响睾丸精子的发生.%Objective To investigate the expression of proliferating cell nuclear antigen (PCNA) and Ki67 in the testis of normal and diabetic mice so as to study the effect of diabetes on the proliferation of spermatogenic cells.Methods Thirty normal male C57BL/6 mice were randomly divided into a control group (n=10) and a DM group (n=20).The diabetic model was established by intraperitoneal injection of streptozotocin into C57BL/6 mouse,while the same volume of citric acid-sodium citrate buffer was injected in the control group mice.The mice were killed three weeks after model establishment,the testes and epididymes were sampled,fixed,then embedded in paraffin,sectioned,stained by HE,and the morphological changes were observed under microscope.The immunohistochemical SP method and image analysis technique were employed to detect and analyze the expression of PCNA and Ki67 in the testis tissue.Results HE staining showed that in the diabetic group,the germ cells at the earlier stage of the spermatogenic cycle were loosely arranged in the seminiferous tubule,the number of cell layers was decreased,and the sperm density in the ductus epididymidis was relatively low.Immunohistochemical staining results showed that the expression of PCNA and Ki67 was significantly lower in the diabetic group than in the normal group (P＜0.01).Conclusion High glucose concentration may impact spermatogenesis by inhibiting the proliferation of spermatogenic cells.
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