背景:何首乌颗粒由何首乌、肉苁蓉、怀牛膝等组成,具有补肝肾、益精血、延年益寿的功效。目的:观察中药何首乌颗粒对D-半乳糖致亚急性衰老大鼠血清睾酮含量及睾丸生精细胞增殖细胞核抗原表达的影响。设计:完全随机分组设计,对照实验。单位:承德医学院基础医学研究所。材料:实验于2003-04/07在承德医学院基础医学研究所完成。选用成年雄性SD大鼠32只。大鼠被随机分为4组:正常对照组、模型组、用药组和阴性对照组,每组8只。方法:①模型组、用药组、阴性对照组大鼠均采用D-半乳糖连续腹腔注射200mg/(kg·d),1次/d,共40d,制作亚急性衰老的大鼠模型。模型组大鼠于造模成功后不再作任何处理;用药组大鼠于造模成功后每天用何首乌颗粒(由灸何首乌、怀牛膝、肉从蓉、茯苓、丹参、淫羊藿六味中药组成,经熬制后每毫升含生药1.5g)灌胃;阴性对照组大鼠在造模成功后每天灌胃同等剂量的生理盐水,时间同用药组大鼠。正常对照组大鼠不造成衰老模型,干预措施同阴性对照组。②血清睾酮水平酶联免疫吸附法检测。增殖细胞核抗原的表达情况采用免疫组化法检测,以细胞核呈桔黄色或棕黄色颗粒为阳性。③计量差异性测定采用t检验。主要观察指标:各组大鼠血清睾酮含量、睾丸生精细胞增殖细胞核抗原的表达比较。结果:大鼠32只均进入结果分析。①血清睾酮水平:模型组大鼠血清睾酮水平明显低于正常对照组犤(0.52±0.14),(1.26±0.32)μg/L,P<0.05犦;用药组明显高于模型组犤(1.16±0.32)μg/L,P<0.05犦。②大鼠睾丸增殖细胞核抗原的阳性细胞率:模型组明显低于正常对照组犤(23.26±3.38)%,(39.15±3.48)%,P<0.05犦;用药组大鼠明显高于模型组犤(37.47±3.18)%,P<0.05犦。③增殖细胞核抗原在生精小管中的分布及变化:各组切片均可见增殖细胞核抗原阳性染色,免疫阳性产物位于细胞核,通常呈棕黄色颗粒,小部分呈弥散状;免疫阳性细胞主要是精原细胞,另外还有少量初级精母细胞、支持细胞和肌样细胞核着色。结论:何首乌颗粒可以逆转衰老大鼠血清睾酮含量及睾丸生精细胞增殖细胞核抗原表达水平的下降,从而延缓生殖功能的退化,具有一定延缓衰老的作用。 Background: Polygonum multiflorum consists of Polygonum multiflorum, Cistanche deserticola, Achyranthes bidentata and so on. It has the effect of supplementing liver and kidney, promoting blood circulation, and prolonging life. Objective: To observe the effect of Heshouwu Granules on serum testosterone levels and the expression of proliferating cell nuclear antigen of testicular spermatogenic cells in subacute aging rats induced by D-galactose. Design: completely random grouping design, control experiment. Unit: Institute of Basic Medicine, Chengde Medical College. MATERIALS: The experiment was performed at the Institute of Basic Medical Sciences, Chengde Medical College from April 2003 to July 2007. 32 adult male SD rats were selected. Rats were randomly divided into 4 groups: normal control group, model group, drug group and negative control group, with 8 rats in each group. METHODS: 1 Rats in the model group, the drug group and the negative control group were treated with D-galactose by continuous intraperitoneal injection of 200 mg/(kg·d) once daily for 40 days to establish a subacute aging rat model. The rats in the model group were no longer treated after the successful modeling; the rats in the medication group were given Heshouwu granules every day after the success of the modeling (by the moxibustion of Polygonum multiflorum Thunb, Achyranthes bidentata, Radix Astragali, Radish, Salvia miltiorrhiza, Epimedium) The rats were given intragastric administration of 1.5 g per ml of crude drug after tanning. Rats in the negative control group were given the same dose of normal saline every day after the establishment of the model. The time was the same as that of the drug-treated group. Rats in the normal control group did not cause the aging model. The interventions were the same as the negative control group. 2 serum testosterone levels detected by enzyme-linked immunosorbent assay. The expression of proliferating cell nuclear antigen was detected by immunohistochemistry and the nuclei were positive for orange or brown-yellow particles. 3 measurement differences using t test. MAIN OUTCOME MEASURES: The levels of serum testosterone and the expression of proliferating cell nuclear antigen in spermatogenic cells of the testes were compared in each group. RESULTS: All 32 rats were involved in the result analysis. 1 Serum testosterone levels: The serum testosterone levels in the model group were significantly lower than those in the normal control group (0.52 ± 0.14), (1.26 ± 0.32) μg/L, P <0.05 犦; the medication group was significantly higher than the model group 犤 (1.16 ± 1) 0.32) μg/L, P<0.05 犦. 2 Rat testicular proliferating cell nuclear antigen positive cell rate: the model group was significantly lower than the normal control group (23.26±3.38)%, (39.15±3.48)%, P<0.05犦; the drug group was significantly higher than the model group. (37.47±3.18)%, P<0.05 犦. 3 Proliferating cell nuclear antigen distribution and changes in seminiferous tubules: positive staining of proliferating cell nuclear antigen can be seen in each group of sections. The immunopositive products are located in the nucleus, usually present as brownish yellow particles with a small part diffuse; immunopositive cells are mainly spermatogonia. Cells, in addition to a small amount of primary spermatocytes, supporting cells, and myoid nuclear staining. Conclusion: Heshouwu granules can reverse the decrease of serum testosterone content and the expression of proliferating cell nuclear antigen in testis spermatogenic cells of aging rats, thereby delaying the degradation of reproductive function and having a certain anti-aging effect.