An Experimental Research On Liucheng Soup Medicated Serum Against MRC-5Cell Aging And Preliminary To Discuss The Mechanism Of This Anti-aging Effect

Objective:Firstly use Liucheng soup in anti-aging research, observing the influence of D-galactose induced aging MRC-5cell’s vitality, the shape, the anti-oxidation damage, apoptosis cycle and apoptosis related protein expression, to provide new theoretical basis for the future anti-aging research of traditional Chinese medicine.Methods:prepare the medicated serum of Liucheng soup, using a dose of80g/1of D-galactose in human embryonic lung fibroblasts cell (MRC-5cells) to prepare aging model. Set up blank serum control group, model group,10%medicated serum,8%medicated serum,6%medicated serum,4%medicated serum,2%medicated serum group (including medicine serum with blank serum dilution),all7groups, through the MTT method to determine the best medicated serum concentration and the best effect of time, according to the results. Set up blank serum control group, model group, high dose group (8%medicated serum), medium dose group (4%medicated serum) and low dose group (2%medicated serum).24hours after cells transfer of culture, join medicated serum according to its groups,placed in (37℃,5%CO2) incubator for48hours, after then take pictures, and collect the supernatant fluid and cells, detection MDA, NO content; T-SOD and GSH-Px activity inside and outside of the cell; Using flow cytometry counting method to detect cell cycle and apoptosis; With Western-blot method detection aging related bax, BCL-2, caspase3protein expression, using SPSS13.0to analysis the data,Results:the model group cells appear flat, bigger, fusion slow, the rest of the group,compared with model group, the oxidase T-SOD and GSH-PX activity enhancement (P<0.05), NO cells and extracellular MDA lower (P<0.05), both inside and outside of the cell, with the most obvious difference in the dose group; Compared to the model:Flow cell counting display that the G1phase cells decreased, S period cell and apoptosis rate increase; Western-blot method shows that promote apoptosis protein caspase3, bax expression is reduced and apoptosis protein bcl-2expression is increased, the most obvious change in media dose group, comparison between groups with analysis of variance, and meet the normality and homogeneity of variance. P <0.05for significant differences.Conclusion:Liucheng soup may by increasing T-SOD and GSH-Px activity of the cell inside and outside, accelerate the death of NO direct and clear extracellular MDA play antioxidant activity; regulate the cell cycle by reduced cells mass that blocked in the G1phase, regulate apoptosis related protein expression, reduce apoptosis rate to play anti-aging effect.