Research Of Anti-Aging Affect And Mechanism Of Ginsenoside Rg1 On Animal Aging Induced By D-Galactose

Objective Stem cell aging is the latest theory of aging, it is important to regulate stem cell in delaying senescence and prevention of senile disease. Neural stem cells (NSCs) in aging and degenerative diseases of the nervous system are closely related. Previous studies have shown that ginsenoside Rg1 was able to add smart and inhibit aging of brain, which is important anti-aging ingredients in ginseng that is traditional Chinese medicine. The reports of Ginsenoside Rgl regulation of NSCs on aging was only observed in our group, but the mechanism during the treatment remains elucidated. To research of anti-aging affect of ginsenoside Rgl on brain aging, we establish the animal and cell aging model induced by D-galactose. The project aims to explain the modern biological mechanism of "theory of air and blood" and "air anti-aging" pathway to regulate NSC aging, provide theoretical and laboratory basis for prevention and control of diseases of nervous system degenerative.Methods1. Research of anti-aging affect of ginsenoside Rgl on animal aging induced by D-galactose.(1) The experimental group:forty male SD rats were randomly divided into normal group, Rgl normal group, brain aging model group and Rgl brain aging model group. Brain aging model group and Rgl brain aging model group were treated with D-galactose(120mg/kg*d) for 42 days by subcutaneous injection; rats of Rgl brain aging model group were treated with Ginsenoside Rgl(20mg/kg*d) 27 days by intra-peritoneal injection after 16 days; rats of Rg1 normal group were treated with saline 42 days by subcutaneous injection and Ginsenoside Rgl 27 days by intra-peritoneal injection after 16days;the same volume saline was given to the normal group by subcutaneous and intra-peritoneal at the same time.(2)Experimental measurement:learning and memory abilities were measured by Morris water maze; The number of senescent cells was detected by SA-β-Gal staining; The level of IL-1 and IL-6 proinflammatory cytokines in hippocampus were detected by ELISA; The activities of SOD, contents of GSH in hippocampus were quantified by chromatometry; The change of telomerase activities and telomerase length were performed by TRAP-PCR and southern blotting assay, respectively. The change of P53 and p19、p21 were performed by western blotting and RT-PCR, respectively.2. Preliminary research of anti-aging affect of ginsenoside Rg1 on cell senescence model induced by D-galactose. Primary cultured neural stem cells were isolated from SD pregnant rats. To assess the types and purity of the cells present in third culture, the expressions of the specific markers-Nesin and flow cytometry nestin in the NSCs.(1) The experimental group:the third cultured cells were dicided into normal group, Rg1 normal group, neural stem cell aging model group and Rg1 neural stem cell model group. Neural stem cell aging model group:treat with D-galactose 10mg/ml for 48h and nomal culture for 48h; Rgl neural stem cell model group:while copying the same treatment as that of nural stem cell aging model group at the begin 48h, then treat with ginsenoside Rgl 8μg/ml; normal group:treat with the same volume of DMEM/F12 96h; Rg1 normal group:treat with the same volume of DMEM/F12 at the beging 48h, then treat with ginsenoside Rgl 8μg/ml.(2) Experimental measurement:Cell proliferation ability were measured by counting the neurospheres; The level of senescent neurospheres was detected by SA-P-Gal staining; The level of MDA and ROS were detected by chromatometry and flow cytometry; he change of p19、p21 and p53 were performed by RT-PCR.Results1. In brain aging model group, the spatial learning and memory capacities were weaken, SA-β-Gal positive granules were increased in section of brain tissue, the activity of antioxidant enzyme SOD and the contents of GSH was decreased in hippocampus, the level of IL-1 and IL-6 was increased in hippocampus, the length of telomere and the activity of telomerase was decreased in hippocampus, and the expression of P53、p19 and p21 was increased.2. In Rgl brain aging group, the spatial learning and memory capacities were enhanced, SA-β-Gal positive granules in section of brain tissue was decreased, the activity of antioxidant enzyme SOD and the contents of GSH was increased in hippocampus, the level of IL-1 and IL-6 in hippocampus was decreased, the length contraction of telomere was suppressed while the change of telomerase activity was increased in hippocampus, and the expression of P53、p19 and p21 was decreased.3. Compared with normal group, the spatial learning and memory capacities were enhanced in Rg1 normal group, SA-β-Gal positive granules in section of brain tissue was decreased in Rg1 normal group, the level of IL-1 and IL-6 in hippocampus was decreased in Rg1 normal group.4. In neural stem cells aging model group, the number of neurospheres were increased, the proportion of SA-P-Gal positive neurospheres were increased, the levels of MDA was increased in cultured and the level of ROS was increased in cell, and the expression of P53、p19 and p21 was increased.5. In Rg1 neural stem cell aging group, the number of neurospheres were reduced, the proportion of SA-β-Gal positive neurospheres were decreased, the levels of MDA was decreased in cultured and the level of ROS was decreased in cell,and the expression of P53、p 19 and p21 was ecreased.6. Compared with normal group,the level of MDA and ROS was decreased in Rgl normal group.Conclusion1. The results indicate that improvement of antioxidant ability, reduction the level of proinflammatory cytokines, regulation of telomerase system and inhibition the expression of senescence-associated gene for rats aging model may be the underlying anti-aging mechanism of Ginsenoside Rgl.2. The results indicate that improvement of antioxidant ability, then inhibit the DNA damage of neural stem cell in brain aging model may be the underlying anti-aging mechanism of Ginsenoside Rgl.