锌-α2糖蛋白和锌指蛋白32在大鼠睾丸及睾周脂肪中的表达

Expression of zinc alpha 2 glycoprotein and zinc finger and BTB domain containing protein 32 in testicle and peritesticular fat of rats

目的:探讨肥胖大鼠睾丸及睾周脂肪中锌α2糖蛋白(ZAG)和锌指蛋白32(ZBTB32)的表达及在睾丸中的定位。方法:清洁级雄性SD大鼠20只,随机分成研究组(高脂饲料喂养)和对照组(普通饲料喂养),12周后成功构建肥胖大鼠模型。采用免疫组织化学法检测ZAG和ZBTB32在睾丸中的定位,采用实时荧光定量PCR和蛋白免疫印迹技术检测ZAG和ZBTB32在大鼠睾丸及睾周脂肪中mRNA和蛋白表达。结果:ZAG和ZBTB32在大鼠睾丸生精细胞胞质中表达;研究组大鼠睾周脂肪中ZAG蛋白的表达量和mRNA相对表达量(0.751±0.030,0.048±0.068)均低于对照组(1.116±0.021,1.000±0.000)(P<0.05);研究组与对照组大鼠睾丸组织中ZAG蛋白表达量(0.616±0.025,0.605±0.033)和mRNA相对表达量(0.820±0.760,1.000±0.000)均无差异(P>0.05);ZBTB32在研究组大鼠睾丸及睾周脂肪组织中蛋白表达量(1.514±0.057,0.376±0.007)与对照组(1.477±0.035,0.390±0.019)比较无差异(P>0.05);ZBTB32在研究组大鼠睾丸及睾周脂肪中mRNA相对表达量(0.994±0.770,0.744±0.650)与对照组大鼠(1.000±0,1.000±0.000)比较无差异(P>0.05)。结论:肥胖大鼠睾周脂肪中ZAG表达显著降低,提示肥胖可能导致ZAG表达异常,进而影响雄性生殖。

Objective:To investigate the expression and localization of zinc alpha 2 glycoprotein and zinc finger and BTB domain containing protein 32(ZBTB32)in testis and peritesticular fat of obese rats.Methods:20 male SD rats with cleanliness grade were randomly divided into control group(normal feed)and study group(high fat feed).After 12 weeks,the obese rat model was established successfully.The localization of ZAG and ZBTB32 in testis was detected by immunohistochemistry.The mRNA and protein expression of ZAG and ZBTB32 in testis and peritesticular fat of rats were detected by real time fluorescence quantitative PCR and Western blot.Results:The relative expression of ZAG protein and mRNA in peritesticular fat of rats in the study group were 0.751±0.030 and 1.116±0.021,which were significant lower than those(0.048±0.068 and 1.000±0))of rats in the control group(P<0.05).The expression of ZAG protein and mRNA in the testis tissue of rats were 0.616±0.025 and 0.820±0.760,which had no significant different from those(0.605±0.033 and 1.000±0)of rats in the control group(P>0.05).The expression of ZBTB32 ZAG protein in the testis tissue and peritesticular fat of rats were 1.514±0.057 and 0.376±0.007,which had no significant different from those(1.477±0.035 and 0.390±0.019)of rats in the control group(P>0.05).The expression of ZBTB32 mRNA in the testis tissue and peritesticular fat of rats were 0.994±0.770 and 0.744±0.650,which had no significant different from those(1.000±0 and 1.000±0)of rats in the control group(P>0.05).Conclusion:The expression of ZAG in peritesticular fat of obese rats decreases significantly,which suggests that obesity may lead to abnormal expression of ZAG and affect the reproduction of male rat.