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目的:探讨营养性肥胖对青春期雄性大鼠睾丸生精细胞凋亡的影响。方法:健康雄性W istar大鼠40只,随机分为两组,每组20只,对照组给予普通饲料喂养,高脂组用高脂、高热量饲料喂养建立营养性肥胖大鼠模型,10周末处死大鼠摘取睾丸。全自动生化分析仪(ACA)检测外周血总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)和低密度脂蛋白胆固醇(LDL-C);光镜观察睾丸组织病理学改变;TUNEL检测睾丸组织凋亡细胞;免疫组化法检测Bc l-2和Bax蛋白的分布和表达;RT-PCR法检测睾丸组织Bc l-2 mRNA和Bax mRNA的表达。结果:高脂组TC、TG、HDL-C和LDL-C(5.17±0.17、1.18±0.09、1.76±0.11、5.08±0.18)较对照组(1.38±0.12、0.39±0.05、0.97±0.07、0.75±0.06)显著升高(P<0.05);高脂组生精细胞凋亡指数(37.17±2.74)较对照组(5.16±0.81)显著升高(P<0.01),凋亡细胞以精原细胞和精母细胞为主;高脂组Bax蛋白和Bax mRNA(153.26±8.74、1.08±0.12)表达较对照组(101.81±6.14、0.37±0.04)明显升高(P<0.01);高脂组Bc l-2蛋白和Bc l-2 mRNA(139.26±7.21、0.46±0.05)表达较对照组(159.37±8.96、1.05±0.11)明显降低(P<0.01)。结论:营养性肥胖诱导大鼠睾丸生精细胞凋亡增多,其机制可能与Bc l-2表达降低和Bax表达升高相关。
Objective: To investigate the effect of nutritional obesity on the apoptosis of spermatogenic cells in testis of adolescent male rats. Methods: Forty healthy Wistar rats were randomly divided into two groups (20 rats in each group). The control group was fed with common diet. The rats in high fat group were fed with high fat and high calorie diet to establish the model of nutritional obesity. Rats were sacrificed and testis removed. Peripheral blood total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) were detected by automatic biochemical analyzer (ACA) TUNEL was used to detect the apoptotic cells in testis tissue. The distribution and expression of Bcl-2 and Bax protein were detected by immunohistochemical method. The expression of Bcl-2 mRNA and Bax mRNA in testis tissue was detected by RT-PCR. Results: Compared with the control group (1.38 ± 0.12,0.39 ± 0.05,0.97 ± 0.07, 0.75 ± 0.06) (P <0.05). The apoptosis index of spermatogenic cells in high-fat diet group (37.17 ± 2.74) was significantly higher than that in control group (5.16 ± 0.81) (P <0.01) (P <0.01). Compared with control group, the expression of Bax protein and Bax mRNA in hyperlipidemia group was significantly higher than that in control group (101.81 ± 6.14,0.37 ± 0.04) (P <0.01) Compared with the control group (159.37 ± 8.96, 1.05 ± 0.11), the expression of l-2 protein and Bcl-2 mRNA (139.26 ± 7.21,0.46 ± 0.05) were significantly decreased (P <0.01). Conclusion: Nutritional obesity induces apoptosis of spermatogenic cells in rat testes. The mechanism may be related to the decrease of Bcl-2 expression and the increase of Bax expression.