| AIM: Study on the correlation between anti-aging effects of crude Lycium barbarum polysaccharides and signal transduction system of Ca2+-CaMMETHODS: Aging model of mice induced by D-galactose was established to study on the anti-aging effects of crude extract Lycium barbarum polysaccharides. For 15 and 30 weeks administration with polysaccharides from Lycium barbarum in mice, the effects of anti-aging pharmacodynamics was evaluated by research learning and memory behavior, anti-fatigue ability, tissue ultramicrostrucre, hematology, enzymology, CaM level, and other physiology biochemical indicators in order to explore the mechanism of anti-aging and analyse the correlation of Ca2+-CaM signal system between them. Calmodulin (CaM) was isolated and purified from bovine brain by Phenyl-Sepharose hydrophobic chromatography column. And its activity was detected by PDE. The molecular weight of CaM was tested by SDS-PAGE and PAGE. Its electrophoresis behavior was observed on with or without of Ca2+, at the same time, and ultraviolet scanning spectrum was identifed. CaM-couple antigens, prepared with cross linking agent of EDC, were used for animal immunization, animal serum immunopotency and levels of CaM was evaluated by ELISA quantitative analysis.RESULTS: Continuous administration with crude Lycium barbarum polysaccharides for 15 and 30 weeks in the anti-aging experiment, degeneration in the mice body abated obviously. In administration group, latent period of searching platform was shorted, escape latency was prolonged, lost times was reduced, leaning and memory ability was reinforced in the test of Moriss Water Maze, Step Down and Step Through, and the anti-fatigue ability was enhanced in running test. The indexes of thymus and spleen degeneration lightened; blood flow rate accelerated; the levels of blood sugar decreased; the levels of MAO-B, AchE, iNOS, MDA decreased, and the activity of SOD, GSH-Px, CAT, ATPase, NO, TNOS increased remarkably in some tissue and serum. Degeneration morphology was lightened in hepatic and nephridial tissue compared with aging model. The purified CaM activated Cyclic nucleotide phosphodiesterase (PDE), and its diagram of electrophoresis showed single strip, molecular weight about 17.3 KDa. The transport ratio of CaM-Ca2+ was faster than CaM-EGTA in SDS-PAGE, while the result was opposite in PAGE. The results of ultraviolet absorption-spectra showed there were 5 peticular absorption peaks in 251,260,267,272 and 283nm. After CaM-couple antigens had been immuned to animals for 6 weeks, the results of double agar diffusion test showed antigen-antibody reaction abviously. Animal serum immunopotency was tested by ELISA and the valency was 1:800, the levels of CaM rise in brain of the administration group.CONCLUSION: The results of anti-aging experiment showed that polysaccharides of Lycium barbarum could enhanced the leaning, memory, anti-fatigue ability, and immune function, and also to safeguard body in physiolology biochemical aspect by removing free radical, improving the activity of antioxidase, protecting hepatic and nephridial tissue, and also displayed anti-aging effects. At the same time, it was avilable that CaM purified by Phenyl-Sepharose hydrophobic chromatography column was prepared for CaM-couple antigen by EDC method. The levels of CaM can be evaluated by means of quantitative analysis of indirect ELISA. The levels of CaM in brain of aging model were decreased, as it probably related to the decline of activity of ATPase which has the ablity of transport Ca2+, and also indirectly resulted in the decline of neurotransmitter in brain, accordingly, affected the ability of learning and memory.
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