Regulation Of Metabotropic Glutamae Receptor β-app Expression In The Role Of Aluminum Neurotoxicity

Objective Aluminum exposure in vitro by cultured neurocytes to study theβ-APP inducedneurocytes loss in aluminum and aluminum neurotoxicity induced by the relation, ofmetabotropic glutamate receptors and the relationship between the neurotoxicity induced byaluminum, andβ-APP and metabotropic glutamate receptors.Methods:①primary culturedneurocytes with AlCl3 ? 6H2O exposed to final concentrations of aluminum were 0mmol / L,0.5mmol / L, 1mmol / L, 2mmol / L. 48h post-contrast microscope observation of the role of cellgrowth state, application Cell Counting Kit-8 group of neurocytes was detected by cell viability,AO-EB and Hoechst staining of aluminum exposure cell survival and apoptosis; flow cytometryAnnexin V-PI double staining quantitative determination of apoptosis rate; using fluorescencequantitative PCR and immunohistochemical detection of neurocytesβ-APP, mGluR1, mGluR3,mGluR7 expression.②The cultured primary neurocytes with 2mmol/L of aluminum wereexposed to different concentrations of metabotropic glutamate receptor agonist or antagonist, todetect the cell viability, flow cytometry, AnnexinV-PI double staining quantification apoptosisrate determined by fluorescence quantitative PCR and immunohistochemical detection ofneurocytesβ-APP, mGluR1, mGluR3, mGluR7 expression.Results:①exposed cell processescould shrink, increasing the cell body rounded, cells decreased, and the cell boundaries unclear;the same time as the pathological changes of Al3 + concentration and exposure time of prolonged;AO-EB and Hoechst staining method shows the aluminum exposed cells morphological changesof apoptosis and necrosis; neurocytes viability test result: The neurocytes activity wassignificantly (P <0.01), 0.5mmol / L, 1 mmol / L, 2mmol / L Al3 + activity of neurocytes dosegroup than 0mmol / L Al3 + group; flow cytometry test results: The group of neurocytes apoptosisrate was significantly (P <0.01), 1mmol / L Al3 + and 2mmol / L Al3 + dose group of neurocyteswas significantly higher than 0mmol / L Al3 + group; fluorescence quantitative PCR andimmunohistochemistry showed that neurocytes: compared with the control, low and mediumdose group mGluR1, mGluR3, mGluR7 gene and protein expression has not yet reachedstatistical significance (P> 0.05), high dose group mGluR1 gene and protein expression increasedsignificantly (P <0.01); compared with the control group, high dose group mGluR3, mGluR7gene and protein decreased significantly (P <0.01); compared with the control group, middle and high dose groupβ-APP gene and protein expression increased significantly (P <0.05);②joinedGroupⅡ,Ⅲmetabotropic glutamate receptor agonist, significantly improved after the neuronsof aluminum-like morphological changes of necrosis, joined the GroupⅡ,Ⅲmetabotropicglutamate receptor antagonist, aluminum can be increased after transfection The morphologicalchanges of cell necrosis; different concentrations of metabotropic glutamate receptor agonists cansignificantly improve nerve cell activity (P <0.05, P <0.01), different concentrations ofmetabotropic glutamate receptor antagonist, significantly reduced cell viability (P <0.05, P<0.01); flow cytometry showed that:Ⅱ,Ⅲgroup of metabotropic glutamate receptor agonistscan significantly reduce the cell apoptosis rate (P <0.01),Ⅱ,Ⅲgroup metabotropic glutamatereceptor antagonist can significantly improve the rate of neuronal apoptosis (P <0.01);fluorescence quantitative PCR and immunohistochemistry results showed that: compared with thecontrol group,Ⅱ,Ⅲgroup by metabotropic glutamate agonist can significantly reduce theneurocytes of aluminum mGluR1, mGluR3, mGluR7, andβ-APP gene and protein expression.Conclusion 1. Aluminum can induce neuronal apoptosis, the extent of apoptosis is closely relatedwith the doses of aluminum. 2. With the increased concentration of aluminum in the increasedexpression ofβ-APP, addingⅡ,Ⅲgroup of metabotropic glutamate receptor agonist, decreasedexpression ofβ-APP, suggesting that:β-APP induced nerve cell loss in aluminum and aluminumchloride neurotoxicity;β-APP and metabotropic glutamate receptors. 3. Aluminum-inducedabnormal expression of mGluRs: mGluR1 expression was increased, mGluR3 mGluR7expression decreased. Tips metabotropic glutamate receptor and nerve toxicity induced byaluminum. 4.Ⅱ,Ⅲgroup of metabotropic glutamate receptor agonists neuroprotective effect.