Objective To investigate the changes of hypothalamus brain neurotransmitter of aging rats and mechanisms of Heshouwuyin(HSWY) on anti hypothalamus aging.Methods The sub-acutely aging model was continually induced by ip injection of D-galactose in rats.Oral HSWY was administered once daily for 60 days after aging induction.The rats were killed by being amputated by head and the hypothalamus was taken.Use the method of ELISA to detect SP,β-EP and GnRH.The concentration of monamine transmitters including noradrenaline(NA),dopamine compared with the normal group(P0.05).HSWY dose-dependently decreased the(DA) and 5-hydroxypatmien(5-HT) in hypothalamus were detected by high-performance liquid chromatography(HPLC),with electrochemical detection.Results GnRH and SP of rats in the model group increased significantly,β-EP,NA,DA,5-HT concentration decreased remarkably determination of GnRH and SP in hypothalamus,and raised the concentration of β-EP NA,DA,5-HT.Conclusion The normal function of hypothalamus is disturbed in aging rats.HSWY has the effects on preventing and delaying senescence of hypothalamus by regulating the disorder of hypothalamus neurotransmitter.

Objective:To investigate the protective effects of HESHOUWU decoction on senile rats' kidney. Methods:The senile model rats were made by abdominal injecting D-galactose;after the model was successfully established,the rats were adminstrated with HESHOUWU decoction.The slices of kidney were made by histological method and stained by HE staining.The morphological changes of kidney were observed under microscope and quantitatively analyzed.Results:The kidney glomerulus number of rats in HESHOUWU decoction group was obviously higher than in model group and approach normal rats(P0.01);the renal capsule breadth and the kidney glomerulus diameter were obviously lower than model group(P0.01).Conclusions: HESHOUWU decoction can improve the morphology of senile rats kidney and can delay renal senility of rats.

目的通过检测p53、Rb、p16、p19、p21在D-半乳糖所致衰老大鼠睾丸组织的表达,探讨松花粉对睾丸组织p53/Rb细胞传导通路的影响。方法采用RT-PCR检测p53、Rb基因在睾丸的表达;Western印迹检测大鼠睾丸磷酸化Rb(p-Rb)、p16、p19、p21蛋白表达。结果模型组睾丸组织p53、Rb基因和p16、p19、p21蛋白的表达明显高于正常组(P<0.01);p-Rb蛋白表达显著低于正常组(P<0.01)。而松花粉能降低p53、Rb基因和p16、p19、p21蛋白的表达提高p-Rb的表达,且明显好于阳性对照组(P<0.05)。结论松花粉通过调节睾丸组织p53/Rb细胞传导通路相关蛋白及基因的表达,起到延缓睾丸组织衰老的作用。

Objective:To investigate the effects of HESHOUWUYIN on expression of TGF-β1 and TIMP-1 in senile rat’s kidney.Methods:The senile rats model was made by abdominal injecting D-galactose and oral administration of HESHOUWUYIN.SP immunohistochemistry was used to observe expression of TGF-β1 and TIMP-1 in kidney.Results:The expression of TGF-β1 and TIMP-1 in kidney of rats in HESHOUWUYIN group were obviously lower than model group(P0.01,P0.05) .Conclusions:HESHOUWUYIN can improve glomerulosclerosis and renal interstitial fibrosis,so has anti-aging effects on kidney of senile rats.

目的观察松花粉对腮腺组织表皮生长因子(EGFR)表达的影响。方法D-半乳糖连续腹腔注射制作亚急性衰老雄性大鼠模型,采用免疫组织化学方法(SP法)观察高、中、低剂量(1400、700、350mg·kg-1·d-1松花粉对衰老的腮腺组织EGFR表达的影响。结果EGFR在腮腺组织广泛表达,D-半乳糖所致亚急性衰老大鼠模型组EGFR的表达明显下降,与正常对照组比较差异显著(P<0.01);而给予松花粉后EGFR的表达明显上升,其中预防中剂量组效果明显。结论松花粉具有改善衰老雄性大鼠腮腺功能紊乱,预防和延缓腮腺组织衰老的作用。

OBJECTIVE:To study the inhibitory effect of Danggui Buxue Tang (DBT), a compound traditional Chinese herbal medicine for supplementing blood, on tumor growth in tumor-bearing mice after inoculation of EL-4 cells, and its immune mechanism as well as its synergic effect in reducing toxicity of cytoxan (CTX).METHODS:Experiment was carried out in tumor-bearing mice after inoculation of EL-4 cells. The mice were randomly divided into four groups after 7 days of the inoculation: untreated group, DBT-treated group [24 g/(kg x d)], CTX-treated group [7.5 mg/(kg x d)] and DBT plus CTX-treated group, with another ten normal mice as control. Inhibitory rate of tumor growth, survival time, immune function and variability of blood cells were measured in the mice during the experiment.RESULTS:After treatment of relevant interventions for 15 days, the tumor in the DBT-treated group, CTX-treated group and DBT plus CTX-treated group grew slower than the untreated group (P<0.05). Murine survival time in the DBT-treated group, CTX-treated group and DBT plus CTX-treated group was lengthened as compared with that in the untreated group (P<0.05). Compared with the untreated group, all kinds of immune indexes in the DBT-treated group and DBT plus CTX-treated group were significantly improved (P<0.05), while the immune indexes in the CTX-treated group were decreased (P<0.05). Compared with the CTX-treated group, all kinds of immune indexes in the DBT plus CTX-treated group were significantly improved (P<0.05).CONCLUSIONS:DBT can enhance the immune function in tumor-bearing mice and the inhibitory effect of DBT on tumor growth is related to the enhanced immune response. DBT can also increase the therapeutic effects and reduce the side effects of CTX.

Objective:To observe the changes of PCNA expression,apoptosis and related factors of spermatogenic cells in the testis of aging rats.Methods:SD rats were used and divided into the normal control group and aging model group at random.The sub-acutely aging model rats were made by injecting D-gal into abdominal cavity continually.SP immunohistochemistry was used to observe the PCNA and telomerase expression,TUNEL to observe the apoptosis, hybridization in situ and Western blotting to observe the p53 gene expression of the spermatogenic cells.Results:Com- pared with normal rats,the PCNA positive cell rate of aging rats decreased,and the number of telomerase positive cells decreased;the apoptotic index increased significantly,and p53 mRNA positive cell rate and p53 protein contents in- creased significantly.Conclusion:The proliferative ability of the spermatogenic cells decreased,apoptosis increased, and the function of testis was reduced in the aging rats.

Objective:To observe the changes of proliferation and apoptosis of cells in testis during different developmental stage of human fetus.Methods:SP immuocytochemistry and TUNEL were used to observe the proliferation and apoptosis of cells in testis of 12~(th) to 32~(nd) week human fetus.Results:In 12~(th) week fetus testis,the cells began apoptosis,and reached peak at 20-32~(nd) week;and the positive products located in nuclei.A great quantity of PNCA positive cells could be observed in 12~(th) week testis,and reached peak at 16-24~(th) week;the PCNA positive cells decreased gradually with age of fetus (P＜0.01).Conclusion:During the development of human fetus,the proliferation and apoptosis of cells in testis express in different level,which indicate that proliferation and apoptosis play an important role in development process of testis.

ObjectiveTo observe the effects of traditional Chinese medicine Tiannianyin on telomerase expression of spermatogenic cells in testis of aging rats.Methods40 SD rats were applied in this study and divided into 4 groups at random: normal group,aging model group,Tiannianyin group,negative control group,and each group had 10 rats.The sub-acutely aging model rats were made by injecting Dgal into abdominal cavity continously.ELISA was used to detect the testosterone content in serum,xanthine oxidase method to detect the superoxide dismutase(SOD) activity in serum,SP immunohistochemistry to observe the telomerase expression of spermatogenic cells in testis.ResultsThe testosterone content and SOD activity in serum of aging rats decreased significantly compared with normal rats(P0.05,P0.01);the telomerase positive cells of aging rats were much less than normal rats.Tiannianyin could inhibit the decreasing of testosterone content and SOD activity in serum significantly(Tiannianyin group compared with aging model group: P0.05,P0.01),and increase the telomerase expression significantly.ConclusionTraditional Chinese medicine Tiannianyin can improve the telomerase expression of spermatogenic cells in testis,and has fairly anti-aging effects.

Objective:To explore the influence of Pine Pollen on anti-oxidative ability in liver of D-gal aging rats. Methods; The sub-acutely aging model rats were made by intraperitoneal injection of D-gal continually. Biochemical methods were used to detect T-AOC, SOD and GSH-PX activities in liver tissue bomogenate of each group. Results- T-AOC, SOD and GSH-PX activities of rats in aging model group significantly decreased compared with normal rats (P0. 01);While the three indexes were remarkably increased after taking Pine Pollen, and there was statistically difference between Pine Pollen and VE (P 0. 01). Conclusions:Pine Pollen could obviously increase the anti-oxidative ability of liver, and has certain anti-aging effects.

目的探讨松花粉的抗下丘脑-垂体-睾丸衰老作用。方法D-半乳糖连续腹腔注射制作亚急性衰老雄性大鼠模型,分别采用放射免疫和酶联免疫法检测血清中黄体生成激素(LH)、卵泡刺激素(FSH)、胰岛素样生长因子1(IGF-1)、睾酮(T)含量以及下丘脑促性腺激素释放激素(GnRH)浓度的变化。结果D-半乳糖所致亚急性衰老大鼠模型组LH、FSH、GnRH均明显上升,IGF-1、睾酮含量明显下降,与正常对照组比较差异显著(P<0.01);而经松花粉给药后LH、FSH、GnRH三者均有显著下降,IGF-1、睾酮含量明显提高。结论松花粉具有改善衰老雄性大鼠下丘脑-垂体-睾丸轴功能紊乱的作用。

目的通过对亚急性衰老大鼠模型血清超氧化物歧化酶（SOD）、丙二醛（MDA）、谷胱甘肽过氧化物酶（GSH-Px）和总抗氧化能力（T-AOC）的测定,观察大鼠精子数量、活动度和存活率的变化,从而探讨松花粉的抗衰老作用。方法D-半乳糖连续腹腔注射制作亚急性衰老大鼠模型,采用生物化学方法检测了各实验组血清中SOD、MDA、GSH-Px和T-AOC的含量;观察了大鼠精子数量、活动度和存活率的变化。结果D-半乳糖所致亚急性衰老大鼠模型组SOD活力、GSH-Px活力和T-AOC均明显下降,MDA含量明显上升,与正常对照组比较差异显著（P＜0.01）;而经松花粉给药后SOD活力、GSH-Px活力和T-AOC三者均有显著提高,MDA含量明显下降;松花粉可提高D-半乳糖所致亚急性衰老大鼠精子的数量、活动度和存活率。结论松花粉可显著提高亚急性衰老大鼠抗氧自由基的能力,提高精子的数量、活动度和存活率,具有延缓衰老的作用。

目的:观察中药天年饮对亚急性衰老大鼠垂体前叶细胞增殖与凋亡的影响.方法:实验于2003-04/07在承德医学院基础医学研究所进行.将40只SD大鼠随机分为4组:正常组、模型组、用药组和阴性对照组,每组10只.模型组、用药组、阴性对照组采用D-半乳糖腹腔注射制作亚急性衰老模型.造模成功后,用药组给予中药天年饮(炙何首乌15 g,怀牛膝15 g,肉苁蓉10g,淫羊藿10 g,丹参25 g,茯苓15g,熬制后每毫升含生药1.5g)灌胃,阴性对照组给予同等剂量生理盐水灌胃.分别采用免疫组化方法和原位末端标记法检测垂体前叶细胞增殖核抗原的表达及凋亡的变化情况.结果:40只大鼠均进入结果分析.①垂体前叶细胞增殖核抗原阳性细胞数量:模型组明显低于正常组[(24.00±3.62),(78.00±3.80)个/视野,(t=81,P＜0.01)];用药组明显高于模型组[(76.00±1.51)个/视野,(t=52,P＜0.01)],并接近正常水平.②垂体前叶细胞凋亡数量:模型组明显高于正常组[(67.00±2.79),(19.00±1.94)个/视野,(t=49.32,P＜0.01)];用药组明显低于模型组[(21.00±3.46)个/视野,(t=41.14,P＜0.01). 结论:天年饮可提高衰老大鼠垂体前叶细胞增殖核抗原表达水平,降低垂体前叶细胞的凋亡指数,说明天年饮可以减弱动物模型垂体功能的退化,具有一定延缓衰老的作用.

Objective: To study the effects of Traditional Chinese Medicine Tiannianyin (TNY) on testestrone level in serum and the PCNA expression in pituitary cell of aging rats. Methods: The model of sub-acutely aging rats were made by injecting D-gal into the abdominal cavity continually. Then the testestrone level in serum and PCNA expression in anterior pituitary cell were detected before and after taking TNY. Results: The testestrone level in serum of aging rats decreased significantly compared with normal rats(P0.05), and the testestrone level increased after taking TNY (compared with aging rats: P0.05). PCNA expression in anterior pituitary cell of aging rats decreased significantly compared with normal rats(P0.01)and after taking TNY it increased significantly (P0.01). Conclusions: TNY has fairly effects on delaying senescence.

目的:观察何首乌颗粒(Heshouwu Granule,HG)对亚急性衰老大鼠睾丸PCNA表达的影响.方法:D-半乳糖(D-gal)连续腹腔注射制作亚急性衰老大鼠模型,检测HG灌胃前后模型大鼠血清睾酮含量及睾丸生精细胞PCNA表达的变化情况.结果:D-半乳糖致亚急性衰老大鼠血清睾酮含量及睾丸PCNA阳性细胞率明显下降,与正常大鼠比较差异显著(P<0.05,P<0.01);经HG灌胃后,二者均升高接近正常水平,与模型大鼠相比有明显差异(P<0.05,P<0.01).结论:HG可提高亚急性衰老大鼠血清睾酮含量及睾丸生精细胞PCNA表达水平,具有一定的延缓衰老作用.

Objective:By studying on the expression of PCNA and NGF in CA 3 area of hippocampus to make out the developmental law of hippocampus . Method: The brain of rats were taken out at postnatal d1,d2,d3,d5,d10,d20,d30 respectively and fixed with 10% formalin, SP immunohistochemistry was used in this study to observe the expression of PCNA,NGF in CA 3 area of hippocampus .Results: The PCNA expression in CA 3 area of hippocampus was the most obvious at postnatal d5:neurons in multiform layer proliferated the most, multiform layer took second place and pyramidal layer the least . The PCNA expression decreased gradually after postnatal d10 and basically ended at postnatal d30. NGF positive cell was not observed in CA 3 area of hippocampus at postnatal d1～d5.The expression of NGF reached the top at postnatal d10 and expressed the most in pyramidal layer and became stable at postnatal d20. Conclusions: The expression of NGF has supporting functions to neurons.

目的:观察天年饮(Tiannianyin,TNY)对亚急性衰老大鼠睾丸组织超氧化物歧化酶(Superoxide dismutase,SOD)的活性及对睾丸生精细胞凋亡的影响.方法:采用D-半乳糖连续腹腔注射制作亚急性衰老大鼠模型,检测TNY灌胃前后大鼠血清睾酮含量、睾丸组织SOD的活性及睾丸生精细胞凋亡的情况.结果:D-半乳糖致亚急性衰老大鼠血清睾酮含量及睾丸组织SOD的活性均明显下降,与正常大鼠比较差异显著(P＜0.05,P＜0.01);经TNY灌胃后二者均升高接近正常水平,与模型大鼠相比有明显差异(P＜0.05,P＜0.01).D-半乳糖致亚急性衰老大鼠睾丸曲细精管凋亡百分率及凋亡细胞阳性率均升高,与正常组比较差异显著(P＜0.05);经TNY灌胃后,二者均降低,与模型大鼠比较有明显差异(P＜0.05).结论:TNY可提高亚急性衰老大鼠血清睾酮含量、睾丸组织SOD的活性及降低衰老大鼠睾丸生精细胞的凋亡指数,具有一定延缓性腺衰老的作用.

目的:观察天年饮(Tiannianyin,TNY)对亚急性衰老大鼠睾丸组织超氧化物歧化酶(SOD)的活性及PCNA表达的影响.方法:采用D-半乳糖连续腹腔注射制作亚急性衰老大鼠模型,检测TNY灌胃前后模型大鼠血清睾酮含量、睾丸组织SOD的活性及睾丸生精细胞PCNA表达的变化情况.结果:D-半乳糖致亚急性衰老大鼠血清睾酮含量、睾丸组织SOD的活性及睾丸生精细胞PCNA阳性率明显下降,与正常大鼠比较差异显著(P＜0.05,P＜0.01);经TNY灌胃后,三者均升高接近正常水平,与模型大鼠相比有明显差异(P＜0.05,P＜0.01).结论:TNY可提高亚急性衰老大鼠血清睾酮含量、睾丸组织SOD的活性及睾丸生精细胞PCNA表达水平,具有一定延缓衰老的作用.

Objective To study the effect of Tiannianyin (TNY) on the activity of superoxide dismutase (SOD) and testosterone level in serum, the PCNA expression and apoptosis of the germ cells in testes of sub-acutely aging rats. Methods Injecting D-gal into abdominal cavity continually made the models of sub-acutely aging rats. Then the SOD and testosterone levels in serum were detected, the PCNA expression and apoptosis of the germ cells in testes were observed before and after treatment with TNY. Results The SOD and testosterone levels in serum, PCNA expression of the germ cells decreased significantly in sub-acutely aging rats compared with normal rats (p0.01, p0.05), the three indexes increased after the treatment with THY. Apoptosis cells in sub-acutely aging rats increased significantly compared with normal rats (p0.05) but decreased after TNY treatment. Conclusions THY has anti-aging effect on sex gland induced by D-gal in rats significantly..