五子衍宗丸通过增强支持细胞自噬改善生精功能

Mechanism of Wuzi Yanzong Pills in improving spermatogenesis by enhancing the autophagy in Sertoli cells

目的:研究五子衍宗丸通过调控睾丸支持细胞自噬以改善大鼠生精功能机制。方法:将25只雄性SD大鼠随机分为对照组,模型组,五子衍宗丸低、中和高剂量组,每组5只。对照组每天早晚给予0.5%羧甲基纤维素钠(CMC-Na)灌胃。模型组每天早上给予雷公藤多苷(20mg/kg),晚上给予0.5%CMC-Na。五子衍宗丸组每天早上给予雷公藤多苷(20mg/kg),晚上分别给予五子衍宗丸低、中、高剂量(0.5、1.0、2.0g/kg)。灌胃30d。计算机辅助精子分析系统测定精子活力、浓度和总数;免疫组织化学染色分析支持细胞自噬标志蛋白轻链蛋白3(LC3)、溶酶体相关膜蛋白2(LAMP-2)的表达;免疫透射电镜观察自噬小体数量。结果:雷公藤多苷可降低精子活力、浓度和总数,降低LC3、LAMP-2的表达(P<0.05,P<0.01),减少自噬小体数量(P<0.05)。与模型组比较,五子衍宗丸中、高剂量组精子活力增加,低剂量组精子活力降低;各剂量组精子浓度和总数增加;中剂量组LC3表达降低,低、高剂量组LC3表达明显增高(P<0.05);各剂量组LAMP-2表达均增高,中、高剂量组显著增高(P<0.05);低剂量组自噬小体数量增多,中、高剂量组数量降低。结论:五子衍宗丸可通过提高支持细胞自噬水平改善生精功能。

Objective: To investigate the mechanism of Wuzi Yanzong Pills(WYP) improving spermatogenesis by regulating the autophagy in Sertoli cells. Methods: Twenty-five male SD rats were randomly divided into five groups, i.e.the control group, the model group, the low-dose WYP group, the medium-dose WYP group and the high-dose WYP group,respectively, 5 rats in each group. The control group was intragastrically administered with 0.5% sodium carboxymethylcellulose(CMC-Na) every morning and evening. The model group was given Triperygium wilfordii multiglucoside(20 mg/kg) every morning and 0.5% CMC-Na at night. The Wuzi Yanzong Pills group was given Triperygium wilfordii multiglucoside(20 mg/kg)every morning, and the Wuzi Yanzong Pills were given low, medium and high doses(0.5, 1.0, 2.0 g/kg) at night. All mice received intragastric gavage for 30 d. Computer-assisted sperm analysis system was used to determine sperm motility, concentration and total number;immunohistochemical staining analysis supported the expression of autophagy marker protein light chain protein 3(LC3) and lysosomal associated membrane protein 2(LAMP-2);immunoradiometric electron microscopy observed the number of autophagosomes. Results: Triperygium wilfordii multiglucoside can reduce sperm motility, concentration and total number, decrease the expression of LC3 and LAMP-2(P<0.05, P<0.01), and lessen the number of autophagosomes(P<0.05). Compared with the model group, the sperm motility increased in the middle and high dose groups of Wuzi Yanzong Pills, and the sperm motility decreased in the low dose group;the sperm concentration and total number increased in each dose group;the LC3 expression in the middle dose group decreased, and the LC3 expression in the low and high dose groups increased significantly(P<0.05);the expression of LAMP-2 increased in each dose group, and the middle and high dose groups increased significantly(P<0.05);the number of autophagosomes increased in the low dose group, and the number in the middle and high dose groups decreased. Conclusion: Wuzi Yanzong Pills can improve spermatogenic function by increasing the level of autophagy in Sertoli cells.