淫羊藿总黄酮延缓衰老的研究

目的：淫羊藿总黄酮(EF)延缓人胚肺二倍体成纤维细胞衰老及神经内分泌、免疫衰老的机制。方法：采用含EF。的血清对人二倍体成纤维细胞2BS细胞株进行处理，观察2BS细胞寿命；采用荧光实时定量PCR法检测p16基因mRNA的表达；ELISA法检测细胞总视网膜母细胞瘤蛋白(Rb)和磷酸化Rb蛋白的含量；TRAP-Hyb单管一步法检测细胞端粒酶活性；端粒限制性片段(TRF)Southem blot法检测2BS细胞端粒长度变化。采用基因芯片技术，检测下丘脑、垂体、肾上腺、脾脏淋巴细胞的基因表达谱。结果：EF能够延长2BS细胞的传代寿命；下调2Bs细胞p16基因mRNA的表达；增加磷酸化Rb蛋白的含量；延缓衰老细胞端粒长度的缩短，并未激活细胞端粒酶活性；EF上调HPAI、轴多种神经递质、激素、细胞因子或其受体表达。结论：淫羊藿总黄酮通过抑制p16基因表达，促进磷酸化Rb蛋白的产生，从而延缓衰老细胞端粒长度的缩短，发挥延缓细胞衰老的作用。EF能上调神经递质受体的表达并通过NEI网络的下行通路激活神经内分泌和免疫系统；通过下调促凋亡、抗增殖基因，上调抗凋亡、促增殖基因的表达，重塑淋巴细胞基因表达的平衡，延缓免疫衰老。

Study on Mechanisms in Delay of Human Aging by Epimedium Flavonnoids

Objective: To investigate the mechanisms of delaying the aging of diploid fibroblasts of human embryoes and lungs and neuroendocrine and immunity by epimediun flavonoids (EF). Method: To handle the 2BS cells of diploid fibroblasts of humans by serum with EF and observe the life of 2BS; to detect the expression of MRNA of p16 gene by the method of fluorescent real - time quantitative PCR; to determine the content of the retinoblastoma protein and the phosphorylase Rb protein of cells by ELISA method; to test the activity of telomerase of cells by the method of simgle - tube and one - step TRAP - Hyb; to measure the change of the length of telomere of 2BS cells by the Southern blot method of telomere ristriction fragment; and to detect the gene expression profile of hypothalamus, pituitary, adrenal, spleen lymphocytes of rats by the technology of gene chips. Result: EF can be able to increase the generation life of 2BS cells, decrease the expression of mRNA of gene p16 of 2BS cells, heighten the content of phosphorylase Rb protein, and delay the reduction of the length of aging telomere cells without activating the activity of the telomerase of cells and upgrade various neuro -transmitters, hormones and cell factors as well as the expression of the receptors in HPAT axis of old rats. Conclusion: By the way of restricting the expression of p16 gene and urging the production of phosphorylase Rb protein to retard the reduction of telomere length of aged cells, EF is able to play the role of delaying the aging of cells, it can also be able to up - grade the expression of the receptors of neuro - transmitters and activate neuro - endocrines and immune system by the down - pathway in NE1 network, and re - establish the balance of the gene expression of lymphocytes and delay the immune aging of cells by the expression of reducing the genes of pro - apoptosis but anti - proliferation and upgrading the genes of anti - apoptosis but pro - proliferation of cells.