目的 探讨锌改善高脂饮食（HFD）诱导的雄性大鼠生殖损伤的作用。方法 将24只雄性SD大鼠随机分为ND组（脂肪提供能量10%）、HFD组和HFD+ZnSO4组（脂肪提供能量60%）,每组8只, ND组给予正常饮食,HFD组和HFD+ZnSO4组给与高脂饮食造模8 w后,ND组和HFD组用等量生理盐水灌胃,HFD+ZnSO4组用25 mg/(kg bw·d)硫酸锌生理盐水溶液灌胃干预8 w;16 w末处死大鼠,取大鼠双侧睾丸、睾周脂肪称重,取新鲜附睾尾部精子检测精子总数、活力,用自动生化分析仪测定血清总胆固醇（TC）、甘油三酯（TG）、低密度脂蛋白胆固醇 (LDL-C)、高密度脂蛋白胆固醇（HDL-C）,用化学发光免疫分析仪检测血清睾酮（T）、雌二醇（E2）、泌乳素（PRL）,电感耦合等离子体质谱法测定睾丸组织锌含量,HE染色观察睾丸组织病理学变化,实时荧光定量PCR和Western blot检测睾丸组织中锌转运体、睾酮合成和凋亡相关基因mRNA和蛋白表达水平。结果 与ND组相比,HFD组体重、睾周脂肪重量、Lee’s指数显著升高（P<0.05）,血清TC、TG、LDL-C水平显著升高,HDL-C显著降低（P<0.05）,精子总数和精子活力显著下降（P<0.05）,血清T水平显著降低（P<0.05）,睾丸出现病理损伤,睾酮合成相关指标3β-HSD1、CYP11A1、LHR、STAR的mRNA和蛋白水平显著降低（P<0.01）,睾丸中总锌含量显著降低（P<0.05）,锌转运蛋白ZIP1、ZIP2显著降低,ZnT1、ZnT8显著升高（P<0.05）,睾丸中抗凋亡指标Bcl-2 mRNA和蛋白水平降低,凋亡指标Bax、Caspase-9、Caspase-12 mRNA和蛋白水平升高（P<0.01）;与HFD组相比,HFD+ZnSO4组体重、睾周脂肪重量、Lee’s指数显著降低（P<0.05）,血清TC、TG、LDL-C水平显著降低,HDL-C显著升高（P<0.05）,精子总数和精子活力显著升高（P<0.05）,血清T水平显著升高（P<0.05）,睾丸损伤改善,睾酮合成相关指标CYP11A1、LHR、STAR的mRNA和蛋白水平显著升高（P<0.01）,睾丸中总锌含量显著升高（P<0.05）,锌转运蛋白ZIP1、ZIP2显著升高,ZnT1、ZnT8显著降低（P<0.05）,睾丸Bcl-2 mRNA和蛋白水平升高,Bax、Caspase-9、Caspase-12 mRNA和蛋白水平降低（P<0.01）。结论 锌可能通过改善生精环境中的锌稳态、增加睾酮分泌和减少细胞凋亡来改善高脂肪饮食诱导的雄性大鼠生殖能力下降。
Abstract
Objective To investigate the effects of zinc supplementation on reproductive injury induced by high fat diet (HFD) in male rats. Methods 24 male SD rats were randomly divided into the ND group (10% calories from fat), HFD group and HFD+ZnSO4 group (60% calories from fat), with 8 rats in each group. ND rats were given a normal diet, and rats in the HFD and HFD+ZnSO4 groups were given a high fat diet. After 8 weeks of modeling, ND group and HFD group were given the same amount of normal saline by gavage, and HFD+ZnSO4 group was given 25 mg/(kg bw·d) zinc sulfate solution by gavage. The rats were killed at the end of 16 weeks. The fat tissues from the testis and peritestis were weighed, and the sperms from the epididymis tail were taken fresh to detect the total sperm number and motility. Serum total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C) were measured by an automatic biochemical analyzer. Serum testosterone (T), estradiol (E2) and prolactin (PRL) were detected by chemiluminescence immunoassay. The content of zinc in testicular tissue was determined by inductively coupled plasma mass spectrometry. The histopathological changes of the testis were observed after HE staining. Real-time fluorescent quantitative PCR and Western blot were used to detect the mRNA and protein expressions of zinc transporters, testosterone synthesis and apoptosis related genes in the testicular tissue. Results Compared with the ND group, body weight, peritestosterone fat weight and Lee's index in the HFD group were significantly increased (P<0.05). Serum TC, TG and LDL-C levels were significantly increased, while HDL-C was significantly decreased (P<0.05). The total sperm count and sperm motility were significantly decreased (P<0.05). Serum T level was significantly decreased (P<0.05). Testicular structure was damaged.The mRNA and protein levels of 3β-HSD1, CYP11A1, LHR and STAR were significantly decreased (P<0.01). The total zinc content in the testis was significantly decreased (P<0.05). The expressions of zinc transporters ZIP1 and ZIP2 were significantly decreased, and ZnT1 and ZnT8 were significantly increased (P<0.05). The levels of anti-apoptosis markers Bcl-2 mRNA and protein in the testis were decreased, while the levels of apoptosis markers Bax, Caspase-9, Caspase-12 mRNA and protein were increased (P<0.01). Compared with the HFD group, body weight, peritestosterone fat weight and Lee's index in the HFD+ZnSO4 group were significantly decreased (P<0.05). Serum TC, TG and LDL-C levels were significantly decreased, while HDL-C levels were significantly increased (P<0.05). The total sperm count and sperm motility were significantly increased (P<0.05). Serum T level was significantly increased (P<0.05). Testicular structure was improved. The mRNA and protein levels of testosterone synthesis related genes CYP11A1, LHR and STAR were significantly increased (P<0.01). The total zinc content in the testis was significantly increased (P<0.05). The zinc transporters ZIP1 and ZIP2 were significantly increased, and ZnT1 and ZnT8 were significantly decreased (P<0.05). The levels of Bcl-2 mRNA and protein in the testis were increased, while the levels of Bax, Caspase-9 and Caspase-12 mRNA and protein were decreased (P<0.01). Conclusion Zinc supplementation ameliorates the reproductive damages induced by high fat diet in male rats by improving zinc homeostasis, increasing testosterone secretion and decreasing sperm apoptosis.
关键词
锌 /
高脂饮食 /
生殖 /
大鼠
Key words
zinc /
high fat diet /
reproduction /
rats
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基金
河北省政府资助临床医学优秀人才培养项目（No.ZF2023175）; 河北省“三三三人才工程”资助项目（No.A202101068）
