参阳胶囊急性毒性及对肾阳虚所致雄性不育大鼠的药效学研究

Acute Toxicity of Shenyang Capsule and Its Pharmacodynamics in Male Sterile Rats Caused by Kidney-yang Deficiency

目的研究参阳胶囊急性毒性及对肾阳虚所致雄性不育大鼠的药效。方法取40只KM小鼠随机分为空白对照组和给药组,以最大剂量1日灌胃2次,给药后连续观察14d,测定最大给药量。取80只SD大鼠,使用腺嘌呤造模法制备大鼠肾阳虚模型,将造模成功的大鼠随机分为6组,正常组、模型组、男宝胶囊给药组、参阳胶囊给药组(低、中、高剂量),连续灌胃给予相对应的药物30d。观察参阳胶囊对腺嘌呤所致肾阳虚大鼠雄性不育的治疗效果采用酶联免疫法(ELISA)测定血清中睾酮(T)的水平,采用免疫印迹法(WB)检测阴茎组织中神经型一氧化氮合酶(nNOS)和内皮型一氧化氮合酶(eNOS),取大鼠的睾丸器官采用苏木素-伊红(HE)染色病理切片、大鼠精液在高倍显微镜下观察精子数量与存活率。结果急性毒性研究中小鼠最大给药量为1.6g/kg。药效学研究中,镜下观察发现与模型组相比,各给药组大鼠睾丸组织中生精小管和精子数量增加;各给药组精液中精子存活率均显著提高(P<0.01);参阳胶囊中剂量给药组大鼠阴茎组织中eNOS、nNOS含量均显著升高(P<0.05);各给药组血清T含量均显著升高(P<0.05,P<0.01)。结论参阳胶囊毒性较低或无毒性,对腺嘌呤所致的肾阳虚模型有治疗作用,能显著改善肾阳虚所致的雄性不育。

Objective To study the acute toxicity of Shenyang Capsule and its pesticide effect in male sterility rats caused by kidney-yang deficiency.Methods 40 KM mice were randomly divided into control group and drug administration group.The maximum dose was gavaged twice a day,and the maximum dose(MFD)was determined after 14 d of continuous observation.80 SD rats were selected and adenine modeling method was used to prepare the rats’kidney-yang deficiency model.The successful modeled rats were randomly divided into 6 groups:normal group,model group,Nanbao Capsule administration group and Shenyang Capsule administration group(low,medium and high doses).The effect of Shenyang Capsule on male sterility induced by adenine in rats with kidney-yang deficiency were observed,enzyme-linked immunosorbent assay(ELISA)was used to detect serum testosterone(T)levels,Western blot(WB)was used to detect the penis in the nerve type of nitric oxide synthase(nNOS)and endothelial nitric oxide synthase(eNOS),the testicular organs of rats were stained with hematoxylin eosin(HE)and semen of rats,the number and survival rate of spermatozoa were observed under high-power microscope.Results Microscopic observation showed that compared with model group,spermatogenic tubules and sperm count in the testicular tissues of rats in each group increased,and sperm count significantly increased;the sperm survival rate of each group was significantly increased(P<0.01);the contents of eNOS and nNOS in the penile tissues of rats in Shenyang Capsule group were significantly increased(P<0.05);the serum T content of each group was significantly increased(P<0.05,P<0.01).Conclusion The toxicity of Shenyang Capsule is low or non-toxic.Shenyang Capsule has therapeutic effect on kidney-yang deficiency model induced by adenine,and can significantly improve male sterility caused by kidney-yang deficiency.