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Effects of Hexachlorobenzene and Macozeb on the Apoptosis of PC-12 Cells

Author: ZuoChunYe
Tutor: ZhangJinSong；XiaoHang
School: Nanjing Medical University
Course: Emergency Medicine
Keywords: EDCs Hexachlorobenzene PC-12 cells Apoptosis p-PLCγ p-ERK1/2 Mancozeb
CLC: R114
Type: Master's thesis
Year: 2011
Downloads: 29
Quote: 0
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Abstract


Objective To investigate the effects of Hexachlorobenzene on the apoptosis of PC-12 cells.Methods: The PC-12 cells were cultured in high glucose Dulbecco’s modified Eagle’s medium （DMEM） and supplemented with 10 % fetal bovine serum, 100μg.ml-1 penicillin, and 100μg.ml-1 streptomycin at 37℃in a 5% CO2 atmosphere. Treated with Hexachlorobenzene （0.1,10,20,100, 200μmol/L） for 24 hours. Cell Counting Kit-8 was used to detected the proliferation and to do toxicity analysis. The morphological changes of PC-12 cells were observed by microscope, and cell apoptosis rates were detected by flow cytometry. The expression of p-PLCγand p-ERK1/2 were determined by Western blot.Results: After treated with Hexachlorobenzene, PC-12 cells had morphological changes and higher apoptosis rates. The changes was evident with the increasing of the dose of Hexachlorobenzene. The 50% inhibiting concentration was 72μmol/L. The apoptosis rate of 200μmol.L-1 was 87.12±3.21%.Conclusions: Hexachlorobenzene could induce apoptosis of PC-12 cells, and the expression of p-PLCγand p-ERK1/2 proteins may play a role in the process of PC-12 cells apoptosis induced by Hexachlorobenzene.Part two:Effects of Mancozeb on the apoptosis of PC-12 cellsObjective To investigate the effects of Mancozeb on the apoptosis of PC-12 cells.Methods The PC-12 cells were maintainedon tissue culture plastic in high glucose Dulbecco’s modified Eagle’s medium （DMEM） and supplemented with 10 % fetal bovine serum, 100μg.ml-1 penicillin, and 100μg.ml-1 streptomycin at 37 in a 5% CO2 atmosphere. Treated with Mancozeb （0,1,10,30,60,120μmol/L） for 24 hours. Cell Counting Kit-8 was used to assess the proliferation and toxicity analysis induced by Mancozeb. The morphological changes of PC-12 cells were observed by microscope, and cell apoptosis rates were detected by flow cytometry to match with the staining of FITC-AnnexinV/PI. The expression of apoptosis associated protein Bcl-2, Bax and p-ERK1/2 were determined by Western blot.Results After treated with Mancozeb, PC-12 cells showed evident morphological changes and higher apoptosis rates, and the changes became more evident as a higher-dose of Mancozeb. The 50% inhibiting concentration is 49.95μmol·L-1,The apoptosis rate of 120μmol·L-1 is 90.46±3.94%. The Bax protein levels were increased and the Bcl-2 protein levels were decreased. The p-ERK1/2 expressions were significantly up-regulated compared with the control group. The grey value of p-ERK1/2 （120μmol/L） is 128.04±2.51 and 178.40±4.04 Conclusions Mancozeb could induce apoptosis of PC-12 cells, and the expression of p-ERK1/ 2 proteins may play a role in the process of PC-12 cells apoptosis induced by Mancozeb.

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