| Objective:To approach the mechanism of glutamate (Glu) and glutamine (Gln) and gamma-aminobutyric acid (GABA) in mediating the antagonism of taurine (Tau) to injury of rats’cerebral ganglion nerve cells induced by manganese (Mn), we applied the primary culture method on cerebral ganglion nerve cells of baby rats.Methods:We took the nerve cells of cerebral ganglion from SPF Wistar rats that were born no more than24hours. And we put the cerebral ganglion nerve cells in primary culture till the optimum state, then the cells of the optimum state were used to enforce following experiments. we selected the concentrations of Mn and Tau through MTT experiments, then we divided the cells that would be cultured for later experiments into eight groups as follows:①Control group;②Tau group;③Mn group of low concentration;④Mn group of moderate concentration;⑤Mn group of high concentration;⑥Preventative group with Mn of low concentration;⑦Preventative group with Mn of moderate concentration;⑧Preventative group with Mn of high concentration. We terminated the culture of each group after24hours handling by treated factors, then used inversely epipolic microscope to observe and take photos. After these we carried out the follow tests:BCA test for detecting concentration of protein that would be referenced in late experiment, HPLC test for detecting the concentrations of3amino acids neurotransmitters both in and out of cells.Results:1、Through MTT experiments we selected Tau of4mmol/L, Mn of0.1mmol/L, Mn of0.2mmol/L and Mn of0.4mmol/L to combine handling factors.2、Compared with control group, Tau group had better shape and cell density, and Mn caused the changes of alive cells’shape and decreased the amount of alive cells. When the concentration of Mn increased, the change of cells also increased. The Preventative groups had more complete form and more alive cells than each corresponding Mn group. The change of cells of Preventative groups increased when the concentration of Mn increased.3、HPLC tests’result were that as follow.①Out of cells:Comparing with Control group, extracellular Glu concentration of Tau group decreased significantly (P<0.01), and extracellular Glu concentrations of every Mn group all increased significantly (P<0.01). Extracellular Glu concentration of Mn group increased gradually when the concentration of Mn increased gradually (P<0.01). Extracellular Glu concentrations of every Preventative group all were lower than each corresponding Mn group (P<0.01). Extracellular Glu concentration of Preventative group increased gradually when the concentration of Mn increased gradually (P<0.01). Comparing with Control group, extracellular Gln concentration of Tau group decreased significantly (P<0.01), and extracellular Gln concentrations of every Mn group all decreased significantly (P<0.01). Extracellular Gin concentration of Mn group decreased gradually when the concentration of Mn increased gradually (P<0.01). Extracellular Gln concentrations of every Preventative group all were lower than each corresponding Mn group (P<0.01). Extracellular Gln concentration of Preventative group decreased gradually when the concentration of Mn increased gradually (P<0.01). Comparing with Control group, extracellular GAB A concentration of Tau group increased significantly (P<0.01). Comparing with Control group, extracellular GABA concentrations of every Mn group all increased, but only the increasing of the highest concentration of Mn in these three groups was significant (P<0.05). Extracellular GABA concentration of Mn group increased gradually when the concentration of Mn increased gradually, but the difference in these three groups was not significant (P>0.05). Extracellular GABA concentrations of every Preventative group all were higher than each corresponding Mn group (P<0.01). Extracellular GAB A concentration of Preventative group increased gradually when the concentration of Mn increased gradually (P<0.01).②In cells:Comparing with Control group, intracellular Glu concentration of Tau group decreased significantly (P<0.01), and intracellular Glu concentration of every Mn group all increased significantly (P<0.01). Intracellular Glu concentration of Mn group increased gradually when the concentration of Mn increased gradually (P<0.01). Intracellular Glu concentrations of every Preventative group all were lower than each corresponding Mn group (P<0.01). Intracellular Glu concentration of Preventative group increased gradually when the concentration of Mn increased gradually (P<0.01). Comparing with Control group, intracellular Gln concentration of Tau group decreased significantly (P<0.01), and intracellular Gln concentration of every Mn group all decreased significantly (P<0.01). Intracellular Gin concentration of Mn group decreased gradually when the concentration of Mn increased gradually (P<0.01). Intracellular Gln concentrations of every Preventative group all were higher than each corresponding Mn group (P<0.01). Intracellular Gln concentration of Preventative group decreased gradually when the concentration of Mn increased gradually (P<0.01). Comparing with Control group, intracellular GABA concentration of Tau group increased significantly (P<0.01), and intracellular GABA concentrations of every Mn group all decreased significantly (P<0.01). Intracellular GABA concentration of Mn group decreased gradually when the concentration of Mn increased gradually (P<0.01). Intracellular GABA concentrations of every Preventative group all were higher than each corresponding Mn group (P<0.01). Intracellular GABA concentration of Preventative group increased gradually when the concentration of Mn increased gradually (P<0.01).③If we consider both in and out of cells at the same time, we can find that as follow. Comparing with Control group, mean concentration of Glu of Tau group decreased significantly (P<0.01), and mean concentrations of Glu of every Mn group all increased significantly (P<0.01). Mean concentration of Glu of Mn group increased gradually when the concentration of Mn increased gradually (P<0.01). Mean concentrations of Glu of every Preventative group all were lower than each corresponding Mn group (P<0.01). Mean concentration of Glu of Preventative group increased gradually when the concentration of Mn increased gradually (P<0.01). Comparing with Control group, mean concentration of Gln of Tau group decreased significantly (P<0.01), and mean concentrations of Gln of every Mn group all decreased significantly (P<0.01). Mean concentration of Gln of Mn group decreased gradually when the concentration of Mn increased gradually (P<0.01). Mean concentrations of Gln of every Preventative group all were lower than each corresponding Mn group (P<0.01). Mean concentration of Gln of Preventative group decreased gradually when the concentration of Mn increased gradually (P <0.01). Comparing with Control group, mean concentration of GABA of Tau group increased significantly (P<0.01). Comparing with Control group, mean concentrations of GABA of every Mn group all increased, but the increasing was not significant (P>0.05). Mean concentration of GABA of Mn group increased gradually when the concentration of Mn increased, but the difference in these three groups was not significant (P>0.05). Mean concentrations of GABA of every Preventative group all were higher than each corresponding Mn group (P<0.01). Mean concentration of GABA of Preventative group increased gradually when the concentration of Mn increased gradually (P<0.01).④When we got mean concentrations of GABA to divide mean concentrations of Glu, we found that as follow. Comparing with Control group, the ratio of Tau group decreased significantly (P<0.01), and the ratio of every Mn group all increased significantly (P<0.01). The ratio of the two higher Mn groups both increased significantly when compared with the lowest Mn group (P<0.01). The ratio of the two higher Mn groups had no difference (P>0.05). The ratio of every Preventative group all were lower than each corresponding Mn group (P <0.01). The ratio of Preventative group increased gradually when the concentration of Mn increased gradually, but the difference in these three groups was not significant (P>0.05).Conclusions:Mn can cause injury of rats’cerebral ganglion nerve cells which were primary cultured, and Tau can antagonize it in some extent. Glu may play an important part in the injury induced by Mn, and GABA may play an important part in the antagonism of Tau to the injury. |
PDF Full Text Request