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The Mechanism of Methylmercury Chloride Induced-apoptosis in C6 Rat Glioma Cell

Author: LeiCuiPing
Tutor: LiZhiChao
School: Jilin University
Course: Health Toxicology
Keywords: Methylmercury C6 glioma Apoptosis Radical Calcium Membrane potential Bcl-2
CLC: R73-3
Type: Master's thesis
Year: 2005
Downloads: 104
Quote: 0
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Abstract


Methylmercury (MeHg) is a well-known environmental pollutant toxic to the nervous tissue, particularly central nervous system (CNS) during development. In the past half century, many researchers have been investigating the mechanism of MeHg-induced neurotoxicity. In addition to a direct effect on neurons, much experimental evidence supports an involvement of the glial component. There are three kinds of glial cells in central nervous system: astrocyte, microglia and oligodendrocytes. MeHg exerts direct, potent, toxic effects on neurons but glial cells seem to be an important target. In this regard, it is suggestive that the largest part of the brain burden is located in astrocytes. In addition, in vitro experiments suggest that astrocytes play a key-modulatory role in MeHg-induced neurotoxicity. MeHg can inhibit Na+/K+ ATPase activity, increase membrane permeability, induce the Na+ uptake and K+ release and cause cell swelling. MeHg can prevent the absorbing of glutamate, aspartate and cystine in astrocyte, stimulating the efflux of excito-amino acids and inducing amino acid-mediated neuronal death. Many studies showed that MeHg can cause the apoptosis of astrocyte but the mechanism is still unkown. In this study, we assessed the methylmercury chloride(MMC) toxicity in C6 glioma cell model and investigated the mechanism of MMC-induced apoptosis. The changes of reactive oxygen species(ROS), mitochondrial trans-membrance potential (MMP),calcium and Bcl-2 protein expressions have been proved in many model to have essential role to apoptosis. Herein, we evaluated the changes of these markers in dose-dependent and time-dependent way. Our results demonstrate that MMC can induce the apoptosis of C6 glioma cell by increasing the production of ROS, damaging MMP, destroying the calcium homeostasis and inhibiting the expression of Bcl-2 protein. Besides, these changes have dose-dependent and time-dependent effects. We choose C6 glioma cell as the glial model because the cell line expresses markers of both oligodendrocytes and astrocyte. Based on our former experiments and references to other researchers, we decided MMC dose range: 1.25μM to 10.0μM. When exposed to 10μM MMC, most of C6 glioma cell showed apoptosis in 24 hour; but when to1.25μM MMC, no evident apoptosis can be observed in 1 hour and 6 hour. We studied the changes of C6 glioma cell exposed to 0.0μM (contol group) 1.25μM (Exp1), 2.5μM (Exp2), 5.0μM(Exp3), 10.0μM (Exp4) from 1 hour to 24 hour.

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