锌转运蛋白在糖尿病雄性大鼠性腺组织中的表达

Expressions of zinc transporters in gonads of diabeticmale rats

目的观察糖尿病雄性大鼠性腺中锌转运蛋白的变化,阐明锌转运蛋白在糖尿病雄性生殖系统中的作用。方法 20只SD雄性大鼠随机分为对照组和糖尿病组(DM),造模成功后处理大鼠,取出睾丸、附睾、精囊和前列腺组织。采用HE染色观察组织的病理变化,应用免疫组化法和Western Blot、RT-PCR检测ZnT1、ZnT8、Zip1和Zip2在两组大鼠上述组织中的定位及表达。结果 (1)HE染色:DM组大鼠睾丸生精小管间质细胞结构发生破坏,出现空泡。在附睾上皮细胞出现了空泡样变化以及部分主细胞的胞核发生溶解破裂,精囊腺腔内褶皱减少,前列腺内壁褶皱增多并且上皮增生。(2)免疫组化染色:与对照组比,DM组睾丸ZnT1、Zip1和Zip2表达降低,ZnT8增高;附睾ZnT1、Zip1和ZnT8表达增加,Zip2降低;精囊腺中ZnT1和Zip1表达降低,精囊腺中ZnT8表达增加。前列腺ZnT1、ZnT8、Zip1和Zip2表达均增加。(3)Western blot和RT-PCR:与对照组比,DM组大鼠ZnT1蛋白和m RNA水平在睾丸、精囊中表达降低,附睾中表达增加;ZnT8蛋白和m RNA水平在睾丸、附睾、精囊、前列腺中均表达增加;Zip1在睾丸、精囊中蛋白和m RNA水平表达降低,在附睾表达增加,在前列腺组织中m RNA水平表达增加;Zip2蛋白水平和m RNA水平在附睾中降低,在前列腺中表达增加,在精囊中蛋白水平增加。结论糖尿病大鼠性腺组织锌转运蛋白表达发生变化,糖尿病导致大鼠生殖系统内锌稳态的失衡。

Objective To observe the changes of zinc transporters in the gonads of diabetic male rats, and clarify their roles in the reproductive system of diabetic male rats. Methods Twenty SD male rats were randomly divided into the control group and the diabetic group(DM). The rats in each group were sacrificed 4 weeks after model establishment, and their testes,epididymis, seminal vesicles and prostate tissues were collected. Pathological changes of the above tissues were observed by HE staining and microscope. The changes in localization and expression of ZnT1, ZnT8, Zip1 and Zip2 in the above tissues were determined by immunohistochemistry and Western Blot and RT-PCR. Results(1) HE staining: The number of cells in the seminiferous tubules decreased in the testis of the DM group, the structure destruction of the interstitial cells and vacuoles occurred in the tissues. There were vacuole-like changes in epididymal epithelial cells, and some main cells exhibited nucleus lysis and rupture. The folds in the seminal vesicle gland cavity decreased, the inner wall of the prostate increased in folds and epithelial hyperplasia.(2) Immunohistochemical staining: Compared with the control group, testis tissue in the DM group presented low expressions in ZnT1, Zip1 and Zip2, but high expression in ZnT8;the epididymis had high expressions in the expressions of ZnT1, Zip1, and ZnT8, but low expression in Zip2;Seminal vesicle glands exhibited low expressions in ZnT1 and Zip1, whereas high expression in ZnT8;Prostate show high expressions in ZnT8, Zip1 and Zip2.(3) Western blot and RT-PCR: Compared with that of the control group, the expressions of ZnT1 at protein and m RNA level in the DM group were all decreased in the testis and seminal vesicles, but its expression was increased in the epididymis;The expressions of ZnT8 at protein and m RNA level were all increased in the testis, epididymis, seminal vesicles, and prostate;the expression levels of Zip1 at protein and m RNA levels were decreased in testis and prostate tissue decreased, but increased in epididymis;The protein level of Zip2 in epididymis and its m RNA level in testis and epididymis were decreased, and the m RNA expression of Zip2 in prostate tissue was increased. Conclusion There were obvious changes in expression of zinc transporter protein in gonad tissue of diabetic rats, and diabetes may results in an imbalance of zinc homeostasis in the reproductive system of rats.