Effect of chaperone-mediated autophagy in MPP(+) -induced SH-SY5Y cells and interventional effect of puerarin / 中国中药杂志
China Journal of Chinese Materia Medica
; (24): 106-112, 2014.
Article
in Chinese
| WPRIM (Western Pacific)
| ID: wpr-319645
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To study the protective effect of puerarin on MPP(+) -induced SH-SY5Y cells by chaperone-mediated autophagy (CMA).</p><p><b>METHOD</b>The Parkinson's disease cell model was established by injuring SH-SY5Y cells with 1 mmol x L(-1) MPP+. The CCK-8 staining was adopted to detect the effect the puerarin of different concentrations on the survival rate of MPP(+)-induced SH-SYSY cells. The autophagosome formation was observed under transmission electron microscope. The AO staining showed the changes in the lysosome activity. RT-PCR was used to detect the changes in Lamp2a and Hsc70 mRNA expressions. The western blotting was adopted to test the expressions of Lamp2a, Hsc70 and alpha-synuclein protein in cells.</p><p><b>RESULT</b>Within the concentration range of 12. 5-50.0 micromol x L(-1), the pretreatment with puerain for 30 minutes could protect the injury of MPP+ in SH-SY5Y cells, and showed a certain dose-effect relationship. The AO staining and electron microscope showed the effect of puerain within the concentration range of 12.5-50.0 micromol x L(-1) on 1 mmol x L(-1) MPP(+)-induced SH-SY5Y cells; autophagosomes emerged in cells, and increased along with the rise in the puerarin dose. The results of the flow cytometry revealed that 50.0 micromol x L(-1) of puerarin could protect against the increase of the ROS level in 1 mmol x L(-1) MPP(+) -induced SH-SY5Y cells and prevent the oxidative injury. The results of RT-PCR and western blotting indicated that puerain within the concentration range of 12.5-50.0 micromol x L(-1) alleviated the MPP(+)-induced SH-SY5Y cell injury, and inhibited the accumulation of alpha-synuclein proteins in MPP(+) -induced SH-SY5Y cells by up-regulating Hsc70, Lamp2a mRNA and protein level.</p><p><b>CONCLUSION</b>Puerarin could protect against the MPP(+) -induced cell injury, whose protective mechanism may be related to the chaperone-mediated autophagy pathway of interventional molecules.</p>
Full text:
Available
Database:
WPRIM (Western Pacific)
Main subject:
Parkinson Disease
/
Pharmacology
/
Piperidines
/
Pyrazoles
/
Autophagy
/
Phagosomes
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Tumor Cells, Cultured
/
Up-Regulation
/
Molecular Chaperones
/
Drug Therapy
Limits:
Humans
Language:
Chinese
Journal:
China Journal of Chinese Materia Medica
Year:
2014
Document type:
Article