Senescent Changes Of The Gonad Axises And Experimental Study On Anti-Aging Effect Of TNY

Senescence is an unavoidable physiological process that is caused by ageing of tissue and cells, declining of organ functions along with the increasing of age. In 1977, Besedevsky advanced the theory of immune-neuroendocrine network and hypothalamus-pituitary-gonad axis (HPG axis) is a important component in the network. The gonad functions decline gradually along with senescence, the function that the central nervous system regulate the HPG axis have also changed, the function of the whole HPG axis decline. Aging model induced by D-galactose (D-gal) is universally accepted, and has been extensively used in the studying field of aging and anti-aging. We plan to observe the senescent changes of D-gal model from the HPG axis point,perfect the study of D-gal aging mechanism; and also study the anti-aging effect of Tiannianyin (TNY), establish theoretical and experimental basis for clinical using of Chinese traditional medicine.A. Senescent changes of the gonad axises1. Objective To observe the senescent changes of the gonad axises of D-gal model.2. Methods80 adult male SD rats were used in this study.80 rats were randomly divided into two groups: ①Normal group, 24 rats, normal SD rats with no disposal; ②Model group, 56 rats, these rats were used to made sub-acutely aging model. After the model was successfully established, they were used to observe the senescent changes and study the anti-aging effect of TNY respectively. ⑴ Establishment of the aging model: D-gal was compounded into 25% solution by physiological saline. The solution was injected into the rats abdominal cavity continually by 200mg/kg/d for 40 days. The weight, behavior, energy and biochemical indexes were observed along with this process to make sure whether the model was successfully established.⑵ The blood was taken from the rats medial angles of the eye, and through centrifugal seperation the serum was obtained, then the following indexes were detected respectively. ① The RT-200C Leidu semi-automatic biochemical analytic instrument was used to detect total cholesterol (TC), triglyceride (TG) and high-density lipoprotein cholesterol (HDL-C) in serum. ② The superoxide dismutase (SOD) in serum was detected by xanthine oxydase method. ③ The testosterone level in serum was detected by ELISA. ⑶ The rats were killed by amputating the head and the hypothalamus was taken. The concentration of monamine transmitters including noradrenaline (NA), dopamine (DA) and 5-hydroxytrypatmien (5-HT) in hypothalamus were detected by high-performance liquid chromatography (HPLC), with electrochemical detection.⑷ The hypothalamus, pituitary and testes were taken out respectively after the rats being infused with 4% polyformaldehyde＋2.5% glutaraldehyde, ultrathin sectioned and observed under transmission electron microscope.⑸ The testes were taken after the rats being infused with 4% polyformaldehyde, embedded in paraffin and sectioned as usual. The thickness was 5-7μm. HE-staining was used to observe the conformation changes of seminiferous tubule; SP immunohistochemistry to observe PCNA and telomerase expression of the spermatogenic cells; TUNEL to observe the apoptosis of spermatogenic cells; in situ hybridization to observe the p53 mRNA expression of spermatogenic cells.⑹ The rats were killed by amputating the head and the testes were taken. Western blotting was used to analyze the p53 protein content of the testes.3. ResultsD-gal could induce a series of changes of HPG axises and relevant biochemical indexes.⑴Common status of the model rats The weight increase of model rats was lower than normal rats; the model rats' hair became yellow and dropped; movement of model rats reduced.⑵ Blood lipidThe TC, TG content in serum of model rats was significantly higher than that of normal rats (P<0.01, P<0.05); while the HDL-C content of model rats was significantly lower than the normal rats (P<0.05).⑶ SOD in serumAfter D-gal was injected , the SOD value in serum of model rats significantly dropped from 327...