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Regulation of Metabotropic Glutamae Receptor β-APP Expression in the Role of Aluminum Neurotoxicity

Author: YangZuo
Tutor: NiuQiao
School: Shanxi Medical
Course: Occupational and Environmental Health
Keywords: Aluminum Nerve cells β-APP Neurotoxicity m GluRs
CLC: R114
Type: Master's thesis
Year: 2010
Downloads: 31
Quote: 0
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Abstract

Objective exposed to aluminum in vitro nerve cell culture studies β-APP in aluminum induced loss of nerve cells in the aluminum-induced neurotoxicity starting to explore metabotropic glutamate receptors aluminum induced neurotoxicity relationship, and β-APP and metabolic glutamate receptors. Methods ① nerve cells in primary culture and AlCl ₃ · 6H ₂ O exposure, so that the aluminum of the final concentration were 0 mmol / L, 0.5mmol / L, 1 mmol / L, , 2mmol / L. Phase contrast microscope after 48h observe the growth state of the cells, detected by Cell Counting Kit-8 group of neurons in the cell viability, AO-EB and Hoechst staining aluminum exposure to cell survival and apoptosis; flow cytometry AnnexinV- PI double staining Quantitative determination of the rate of apoptosis; fluorescence quantitative PCR and immunohistochemical detection of beta-APP nerve cells of mGluR1 of mGluR3, mGluR7 of expression. The ② 2mmol / L aluminum exposure in primary cultures of nerve cells with different concentrations of metabotropic glutamate receptor agonists or antagonists detect vitality of cells, flow cytometry Annexin V-PI double staining Quantitative determination of the rate of apoptosis, quantitative PCR and immunohistochemical detection of β-APP of the nerve cells of mGluR1, of mGluR3, mGluR7 expression. Results ① exposure allows cells protruding atrophy, rounded cell body increases, the decrease in the number of cells and cell ill-defined; pathological changes Al3 concentration and exposure time and aggravation; AO-EB and Hoechst staining the morphological changes of of aluminum exposure apoptosis and necrosis; nerve cell viability test results: the group of nerve cell viability difference was statistically significant (P <0.01), 0.5 mmol / L, 1 mmol / L, 2 mmol / L Al ^{3 dose group of neurons in the vitality below 0mmol / L Al ^{3 group; flow cytometry test results: group of nerve cell apoptosis rate difference was statistically significance (P <0.01), 1mmol / L Al ^{3 and 2 mmol / L Al ^{3 dose group of nerve cells apoptosis rate significantly higher than 0mmol / L Al ^{3 group; fluorescence quantitative PCR and immunohistochemical detection of nerve cells results show that: Compared with the control group, low, middle dose group of mGluR1 of mGluR3, mGluR7 gene and protein expression has not yet reached statistical significance (P gt; 0.05), the mGluR1 the high dose group gene and protein expression increased, the difference was statistically significant (P lt; 0.01);, high dose group mGluR3, mGluR7 gene and protein expression decreased compared with the control group, the difference was statistically significant (P lt; 0.01); compared with the control group, the β-APP gene and protein expression in the low-dose group not reach statistical significance (P gt; 0.05) in the high dose group β-APP gene and protein expression. increased, the difference was statistically significant (P lt; 0.05); ② join Ⅱ, Ⅲ metabotropic glutamate receptor agonist can significantly improve the nerve cells exposed to aluminum necrosis morphological changes, adding Ⅱ, Ⅲ group metabotropic glutamate receptor antagonists can increase the morphological changes of the cells exposed to aluminum necrosis; different concentrations of metabotropic glutamate receptor agonist can significantly improve nerve cell vitality (P lt; 0.05, P lt; 0.01) different concentrations of metabotropic glutamate receptor antagonists can significantly reduce cell viability (P lt; 0.05, P lt; 0.01); measured by flow cytometry results showed that: Ⅱ, group Ⅲ metabotropic glutamate receptor agonist can dramatically reduce the nerve cell apoptosis rate (P lt; 0.01) II the group Ⅲ metabotropic glutamate receptor antagonist can significantly improve nerve cell apoptosis rate (P lt; 0.01); quantitative PCR and immune staining showed that: Compared with the control group exposed to aluminum, II, group Ⅲ metabotropic glutamate receptor agonist can significantly reduce the the the dyed aluminum nerve cells of of mGluR1 of mGluR3, mGluR7 and β-APP gene and protein expression. Conclusions aluminum induced neuronal apoptosis, closely related to the degree of apoptosis aluminum dose. With the increased concentration of the dye aluminum increased expression of β-APP, adding Ⅱ, Ⅲ metabotropic glutamate receptor agonist β-APP decreased expression: β-APP in the loss of nerve cells induced by aluminum and aluminum chloride neurotoxicity; β-APP metabotropic glutamate receptors. The aluminum induced mGluRs abnormal expression: of mGluR1 expression is elevated, mGluR3mGluR7 decreased expression. Tip metabotropic glutamate receptor neurotoxicity induced by aluminum. Ⅱ, Ⅲ metabotropic glutamate receptor agonists have neuroprotective effects.}}}}}

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