The Effect On The Proliferation Of Rats’ Olfactory Ensheathing Cells That Primary Cultured In Various Concentrations Lycium Bararum Polysaccharides And Astragalus Injection

The Purity and Identification of Olfactory Ensheathing CellsObjective To explore the culture and the identification of olfactory ensheathing cellsthat from the rats’ olfactory bulbs, and the concentration of olfactory ensheathing cells.Method In this experiment, we use the Nash differential attachment to purify ratolfactory ensheathing cells in vitro which from rat’s olfactory bulbs. Then we mark thespecificity antigen--Low-affinity nerve growth factor receptor p75with specificity antibodylabeled green immunofluorescence and make the nucleus blue with the help of DAPI. Countthe greens and blues, and their ratio is the right result we ever want.Result The maturity olfactory ensheathing cells adhered to the wall, and most of themare bipolar or tripolar with long and thin processes, high refraction, fruity. Seven days later,the main cells are bipolar--OECs, and their growth momentum is direction. After purity, wecan see the major cells are olfactory ensheathing cell-like cells with the help of microscope.When we mark them with immunofluorescence, there were65percent OECs in the wholecells.Conclusion The method that Nash differential attachment to purify rat’s olfactoryensheathing cells in vitro which from rat’s olfactory bulbs was available, and the propotion of olfactory ensheathing cells is at least65percent. The Effect on the Rat’s Olfactory Ensheathing Cells in VariousConcentrations Lycium Bararum Polysaccharides and AstragalusInjectionObjective To explore the proliferation of rats’ olfactory ensheathing cells in vitro withlycium bararum polysaccharides and astragalus injection and to explore the bestconcentrations.Method We use Nash differential attachment to purify rat’s olfactory ensheathing cellsin vitro, then put them in different plates. There are nine group: O is the blank that the culturemedium without fetal bull serum, and the rest are experiment groups--A、B、C、D are thegroups of lycium bararum polysaccharides, and the concentration are as follow: A is20μg/ml, B is50μg/ml, C is100μg/ml and D is200μg/ml; E、F、G、H are the groups ofastragalus injection, E is0.05g/ml, F is0.1g/ml, G is0.2g/ml and H is0.5g/ml. Then we useabove cells to p75immunofluorecent staining, CCK-8test and use the supernate of cellsculture medium to make ELISA test in order to know the content of NGF and BDNF(shownwith absorbance values). The date are analyzed with SPSS18.0software, using a one-wayanalysis of variance for the date of normal distribution, otherwise, the Kruskal-Wallis H testwas used. Results are expressed as mean±SD, and p value <0.05are considered statistucallysignificant.Result①The groups of lycium bararum polysaccharides: result of ELISA: A、B、C、D groups compare with O group, p value <0.05, and C group compare with A、B、D groups, pvalue <0.05. The result of CCK-8: A、B、C、D groups compare with O group, p value <0.05,and C group compare with A、B、D groups, p value <0.05. The result of immunofluorecent staining: A、B、C、D groups compare with O group, p value <0.05, and C group compare withA、B、D groups, p value <0.05.②The groups of astraglus injection: the results of ELISA: E、F、G、H groups compare with O group, p value <0.05, and F group compare with E、G、Hgroups, p value <0.05. The result of CCK-8: E、F、G、H groups compare with O group, p value<0.05, and F group compare with E、 G、 H groups, p value <0.05. The result ofimmunofluorecent staining: E、F、G、H groups compare with O group, p value <0.05, and Fgroup compare with E、G、H groups, p value <0.05.Conclusion Olfactory ensheathing cells can be proliferated with Lycium bararumpolysaccharides and astraglus injection, and the best concentrations are100μg/ml and0.1g/ml respectively.