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目的建立小鼠1型糖尿病模型,研究糖尿病早期时睾丸中睾酮合成关键酶的变化。方法雄性成年鼠20只,随机分为对照组10只、糖尿病模型组10只。通过ip链脲佐菌素(150 mg/kg)制备糖尿病模型,成功制备糖尿病模型后4周处死小鼠,取血清和睾丸组织备用。实时荧光定量PCR(q RT-PCR)法检测睾丸组织中促黄体生成素受体(LHR)、类固醇激素合成灵敏调节蛋白(St AR)、细胞色素P450胆固醇侧链裂解酶(P450scc)、3β-羟基类固醇脱氢酶VI型(3β-HSD6)、17a-羟基化酶/17,20-裂解酶I型(P450c17a1)及17β-羟基类固醇脱氢酶III型(17β-HSD3)相应m RNA(Lhcgr、Star、Cyp11a1、Hsd3b6、Cyp17a1和Hsd17b3)含量,酶联免疫吸附法测定血清中睾酮和黄体生成素(LH)含量、睾丸组织中3β-HSD1、P450c17a1及17β-HSD3酶活性。结果与对照组比较,4周糖尿病小鼠血清中睾酮和LH含量显著降低(P<0.05);睾丸组织的Lhcgr、Star、Cyp11a1、Hsd3b6、Cyp17a1和Hsd17b3 m RNA含量显著降低(P<0.05、0.001);3β-HSD1、P450c17及17β-HSD3酶活性显著降低(P<0.05、0.01)。结论早期糖尿病小鼠睾丸中睾酮合成关键酶的表达显著降低。
Objective To establish mouse model of type 1 diabetes to study the changes of testosterone - synthesizing key enzyme in early stage of diabetes mellitus. Methods Twenty male adult mice were randomly divided into control group (n = 10) and diabetic model group (n = 10). The diabetic model was induced by ip streptozotocin (150 mg / kg). After 4 weeks of successful diabetic model, mice were sacrificed and the serum and testicular tissue were taken for further study. Real-time quantitative PCR (q RT-PCR) was used to detect the expression of LHR, St AR, P450scc and 3β- Hydroxysteroid dehydrogenase type VI (3β-HSD6), 17a-hydroxylase / 17,20-lyase type I (P450c17a1) and 17β-hydroxysteroid dehydrogenase type III (17β-HSD3) , Star, Cyp11a1, Hsd3b6, Cyp17a1 and Hsd17b3). Serum levels of testosterone and luteinizing hormone (LH) and the activities of 3β-HSD1, P450c17a1 and 17β-HSD3 in testis tissue were determined by enzyme linked immunosorbent assay. Results Compared with the control group, the levels of testosterone and LH in the serum of 4-week diabetic mice were significantly decreased (P <0.05), and the contents of Lhcgr, Star, Cyp11a1, Hsd3b6, Cyp17a1 and Hsd17b3 m RNA in testis tissues were significantly decreased (P <0.05, ). The activities of 3β-HSD1, P450c17 and 17β-HSD3 were significantly decreased (P <0.05, 0.01). Conclusion The expression of key enzymes of testosterone synthesis in testes of early diabetic mice was significantly decreased.