天年饮干预亚急性衰老大鼠模型的性腺功能(英文)

Intervention of gonad function with tian nian yin in acute senile rat model

背景:随着机体衰老性腺功能逐渐降低,中国传统医学认为精气虚弱是导致机体衰老的主要原因。目的:观察中药天年饮对亚急性衰老大鼠血清睾酮含量、睾丸组织超氧化物歧化酶的活性及睾丸生精细胞凋亡的影响。设计:随机对照实验。单位:承德医学院人体解剖学教研室。材料:实验于2003-04/07在承德医学院基础医学研究所进行。选择成年雄性SD大鼠40只,随机分为4组,每组10只。模型组、用药组、对照组采用D-半乳糖连续腹腔注射200mg/(kg·d)制备亚急性衰老动物模型,1次/d,共40d,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天年饮灌胃(天年饮由灸何首乌、怀牛膝、肉从蓉、茯苓、丹参、淫羊藿6味中药组成,经熬制后含生药1.5g/mL),1次/d,共30d;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。方法:各组大鼠血清中睾酮的含量采用酶联免疫吸附方法检测,睾丸组织中超氧化物歧化酶的活性采用黄嘌呤氧化酶法检测,睾丸生精细胞的凋亡情况采用免疫组化法观察。主要观察指标:①各组大鼠血清中睾酮的含量。②睾丸组织中超氧化物歧化酶的活性。③睾丸生精细胞的凋亡情况。结果:40只大鼠均进入结果分析。①血清中睾酮的含量:模型组大鼠血清睾酮含量低于正常组犤(0.52±0.15)μg/L,(1.26±0.32)μg/L,(t=3.004,P<0.05)犦,用药组高于模型组犤(1.16±0.32)μg/L,(0.52±0.15)μg/L,(t=2.321,P<0.05)犦。②睾丸组织超氧化物歧化酶的活性:模型组明显低于正常组犤(107.22±9.33)kNU/g,(147.73±11.9)kNU/g,(t=13.339,P<0.01)犦,用药组明显高于模型组犤(141.05±14.57)kNU/g,(107.22±9.33)kNU/g,(t=9.970,P<0.01)犦。③生精细胞的凋亡情况:模型组凋亡生精小管百分率、凋亡阳性细胞率均明显高于正常组犤(53.95±2.02)%比(34.21±2.10)%,(8.67±0.80)%比(3.86±0.52)%,(χ2=7.9,P<0.01,χ2=5.01,P<0.05)犦;用药组明显低于模型组犤(35.33±2.18)%比(53.95±2.02)%,(4.68±0.74)%比(8.67±0.80)%,(χ2=7.02,P<0.01,χ2=3.96,P<0.05)犦。结论:衰老模型睾丸生精细胞凋亡增多,经天年饮灌胃后可以使凋亡细胞阳性率明显下降,说明天年饮可以清除体内过多的氧自由基,减少凋亡发生,改善动物模型的生精功能。

BACKGROUND： In follow of the gradual decrease of senile gonad function in the body, traditional Chinese medicine views that deficiency of essential qi is the chief reason of body senility. OBJECTIVE： To observe the effects of tian nian yin on serum testosterone content, superoxide dismutase activity in testicular tissue and apoptosis of testicular seminiferous cell in subacute senile rats. DESIGN： Randomized controlled experiment. SETTING： Human Anatomy Department of Chengde Medical College. MATERIALS： The experiment was performed in Institute of Basic Medicine of Chengde Medical College from April to July 2003, in which, 40 SD male rats were employed and randomized into 4 groups, 10 rats in each group. In model group, medicinal group and the control, D-galactose was injected 200 mg/（kg·d） to prepare subaeute senile animal model, once a day, continuously for 40 days. No any management was given in model group after modeling success. In medicinal group, gastric infusion with tian nian yin [composed of zhi heshouwu （Radix Polygoni Mutiflori Preparatea）, huainiuxi （Radix Aehyranthis Bidentatae）, roucongrong （Herba Cistanehis）, fuling （Poria）, danshen （Radix Salviae Miltiorrhizae） and yinyanghuo （Herba Epimedii）, it was decocted till 1.5 g/ml, with raw herbs contained] was administrated once a day, totally for 30 days after modeling success. In the control, gastric infusion was administrated with physiological saline of same usage as the medicinal group after modeling success. INTERVENTIONS： In every group, enzyme linked immunosorbent assay （ELISA） was used to determine testosterone content in serum. Xanthine oxidase assay （XOD） was used to determine superoxide dismutase activity in testieular tissue and immunohistochemical method （1HC） was used to observe apoptosis of testicular seminiferous cell. MAIN OUTCOME MEASURES： ① Testosterone content in serum. ② Superoxide dismutase activity in testicular tissue. ③ Apoptosis of testicular seminiferous cell. RESULTS： Forty rats all entered result analysis. ① Testosterone content in serum： Testosterone content in model group was lower than normal group [（0.52±0.15） μg/L, （1.26±0.32） μg/L, （t=3.004, P 〈 0.05）], and that in medicinal group was higher than model group [（1.16±0.32） μg/L, （0.52±＇0.15） μg/L, （t=2.321, P 〈 0.05）]. ② Superoxide dismutase activity in testicular tissue： Superoxide dismutase activity in model group was lower remarkably than the normal group [（107.22±9.33） kNU/g, （147.73±11.90） kNU/g, （t=13.339, P 〈 0.01）], and that in medicinal group was higher remarkably than model group [（141.05±14.57） kNU/g, （107.22±9.33） kNU/g, （t=9.970,P 〈 0.01）]. ③ Apoptosis of testieular seminiferous cell： Percentage of apoptotic seminiferous tubule and apoptotie rate of positive cell in model group were higher remarkably than normal group respectively [（53.95 ±2.02）%compared with （34.21±2.10）%, （8.67±0.80）% compared with （3.86±0.52）%, （X^2=7.9, P 〈 0.01, X^2=5.01, P 〈 0.05）], and those in medicinal group were lower remarkably than model group[（35.33±2.18）% compared with （53.95±2.02）%, （4.68±0.74）% compared with （8.674-0.80）%, （X^2=7.02, P 〈 0.01, X^2=3.96, P 〈 0.05）]. CONCLUSION： The apoptosis of testicular seminiferous cell was in creased in senile model group. Tian nian yin in gastric infusion decreased apoptotic rate of positive cell. It is explained that tian nian yin eliminates surplus oxygenic free radical, decreases apoptosis and improves seminiferous function in animal model.