2020 Fiscal Year Research-status Report
Does the combination of nicotine and ethanol facilitate or block dopaminergic neuron damage in Parkinson's disease models?
Project/Area Number |
19K10687
|
Research Institution | Kagawa University |
Principal Investigator |
モストファ ジャーマル 香川大学, 医学部, 助教 (50418802)
|
Co-Investigator(Kenkyū-buntansha) |
塚本 郁子 香川大学, 医学部, 寄附講座教員 (10183477)
|
Project Period (FY) |
2019-04-01 – 2022-03-31
|
Keywords | Ethanol / Nicotine / MPP+ / PC12 |
Outline of Annual Research Achievements |
In vitro study for the effects of ethanol (EtOH), nicotine or their combination and acetaldehyde (AcH) on lactate dehydrogenase (LDH) in cell culture models of Parkinson’s diseases. I.EtOH concentration was measured in PC12 cells after 24 and 48 hr ethanol exposure by head-space GC. The final concentration of EtOH in the culture media was approximately 99 %. II.PC12 cells were maintained in DMEM 10% fetal bovine serum, 5% horse serum, and antibiotics. The culture medium was changed consisting of serum free DMEM and saline with additives (MPP+, EtOH, nicotine, AcH and COA-Cl). The experimental groups were as follows: Untreated control, PD model (treated with 250 uM MPP+), EtOH (10, 20, 50, or 100 mM), AcH (10, 20, 50, or 100 uM), nicotine (0.5, 1, 2, or 4 mM) alone or combination with EtOH (100 mM) and nicotine (4 mM) in MPP+ model. Cell viability was measured with the water-soluble tetrazolium salt (WST)-8 assay. The cell viability of the control group (serum free DMEM) was set to 100%, and the cell viability of the other groups was compared with that of the control group. MPP alone for 72 h markedly decreased the cell viability compared to control. However, exposure of EtOH, AcH and nicotine did not cause a significant change of viability as well as the LDH activity as compared with those in MPP+. In addition, MPP+ alone for 24 h markedly increased the LDH release, while treatment with COA-Cl attenuated this LDH release. These results suggest that COA-Cl protects against MPP+ -induced cell damage.
|
Current Status of Research Progress |
Current Status of Research Progress
4: Progress in research has been delayed.
Reason
Due to the construction of animal facility which began in May 2020. The neurotoxin MPTP will be required to create Parkinson's disease (PD) in mice. And our lab has no enough facilities to provide this experiment.
|
Strategy for Future Research Activity |
Role of systemic administration of nicotine, ethanol and or acetaldehyde on dopamine release, TH phosphorylation at Ser-31 and Ser-40, PKs, PARP-1 and caspase in the the mouse brain of PD models.
|
Causes of Carryover |
Most part of the experiments during last year were not able to perform due to the construction of animal facility.
|