Tongluojiu'nao Brain Injection Stem Cell Proliferation And Differentiation Of Embryonic Rat Cortical Neurons In Vitro Experimental Study

Neural stem cells have the ability of potential differentiation, that is orientation differentiation under some conditions. When the adult nerve tissue is destroyed or degenerated, it is possible to replace the injured nerve tissue or cells with the homologous nerve tissue or cells produced through the orientation inducement of neural stem cells. In this study we try to investigate the influences of Tong-luo-jiu-nao Injection on neural stem cells, extend the basic theories and methods of modern medical science which promoting tissue regeneration after removing blood stasis in treatment of brain disease, and hoped that through the in-depth study of neural stem cells lay the groundwork for alternative treatment.1 Neural stem cell culture and the cell multiplication effect of Tong-luo-jiu-nao Injection on neural stem cells using MTS/PMS ColorimetryWe selected cortical cells of E17 embryonic rat, and investigated the effects of Tong-luo-jiu-nao Injection on the cell proliferation by primary cell culture, passage, immunofluorescence and MTS/PMS colorimetry. The results showed that cells have Nestin expression and the basic attributes (properties) of neural stem cells which belong to the stem cells in the central nervous system. We adopted MTS/PMS colorimetry to measure the cell proliferation effects of Tong-luo-jiu-nao Injection different in concentrations at the same time point and those in the same concentration at different time points. The results showed that Tong-luo-jiu-nao Injection promotes cell proliferateion at 0.77-123.2μg/ml, the most at 15.4μg/ml, and has inhibitory action on the proliferation when the concentration is more than 492.8μg/ml. When the concentration is 246.4μg/ml, there is no significance between the treatment and the control group which possibly caused by the reason that the promoting and prohibiting effects are on the balance. Until the Tong-luo-jiu-nao Injection administrated for 6h, can it promote the neural stem cells to proliferate.2. Effects of Tong-luo-jiu-nao Injection on the proliferation and differentiation of the neural stem cellsIn this study, we adopted the immunofluorescence double labeling and RT-PCR to investigate the effects of Tong-luo-jiu-nao Injection on the proliferation, differentiation and self-renew of the neural stem cells. The results shown that compared with the control group, the area density, optical density and numbers of the Nestin/Brdu,Nestin/Vimentin,Nestin/Tuj-1 positive cells in the Tong-luo-jiu-nao Injection group are much higher with significance. The mRNA bands of the three genes of Tuj-1,Vimentin,Nestin are in the range of 200-300bp, nearing 300bp. The bands in the treatment group are wider than those in the control group. The value of the integral optical density of these three...