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Amiloride protects PC12 cells from MPP+ induced injury via autophagy-lysosome pathway |
ZHANG Run-ping, LI Jian-ping |
(Department of Neurology,Suzhou New District People′s Hospital,Suzhou Jiangsu 215129,China) |
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Abstract Objective: To evaluate the neuroprotective effects of amiloride; a non-selective blockers of acid-sensing ion channels, on PC12 cells and to view the influence of autophagy-lysosome pathway (ALP). Methods: PC12 cells were tested with methy-phenylpyridi(MPP+) ,while giving amiloride as interventiong:The cells viability was analyzed by MTT assay; The cells injury was assessed by lactate dehydrogenase (LDH) assay;Flow cytometry was used to study the apoptotic;Western blot analysis was used to study the autophagic mechanisms. Results: MPP+ medium treatment resulted in significantly lower survival rate,the supernatant of LDH leakage was significantly higher,the apoptosis rate increased,the expression of LC3-Ⅱ was inhibited while induced more expression of LAMP2a;but amiloride effectively improved the cell survival rate,reduced LDH leakage rate,inhibited apoptosis,and upregulated LC3-Ⅱ protein,which is associated decrease expression of LAMP2a. Conclusion: Amiloride can protect PC12 cells against MPP+-induced cell death by autophagic,through inhibition of acidsensing ion channels activity.
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Received: 13 May 2014
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