title: [Effect of chaperone-mediated autophagy in MPP(+) -induced SH-SY5Y cells and interventional effect of puerarin].
abstract: OBJECTIVE
  To study the protective effect of puerarin on MPP(+) -induced SH-SY5Y cells by chaperone-mediated autophagy (CMA).
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  Puerarin could protect against the MPP(+) -induced cell injury, whose protective mechanism may be related to the chaperone-mediated autophagy pathway of interventional molecules.
authors.forename: Xun-Cui
authors.initials: XC
authors.lastname: Wang
authors.forename: Xiu
authors.initials: X
authors.lastname: Wang
authors.forename: Qing-Lin
authors.initials: QL
authors.lastname: Li
mesh_terms: D001343:Autophagy / Q000187:drug effects* / Q000235:genetics; D050883:HSC70 Heat-Shock Proteins / Q000235:genetics; D006801:Humans; D007529:Isoflavones / Q000494:pharmacology*; D052119:Lysosomal-Associated Membrane Protein 2 / Q000235:genetics; D018832:Molecular Chaperones / Q000235:genetics*; D010300:Parkinson Disease / Q000188:drug therapy / Q000235:genetics; D010588:Phagosomes / Q000187:drug effects / Q000235:genetics; D010880:Piperidines / Q000494:pharmacology*; D011720:Pyrazoles / Q000494:pharmacology*; D014407:Tumor Cells, Cultured; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics
chemical_list: C524478:1,3-bis(4-hydroxyphenyl)-4-methyl-5-(4-(2-piperidinylethoxy)phenol)-1H-pyrazole; D050883:HSC70 Heat-Shock Proteins; C494371:HSPA8 protein, human; D007529:Isoflavones; C497811:LAMP2 protein, human; D052119:Lysosomal-Associated Membrane Protein 2; D018832:Molecular Chaperones; D010880:Piperidines; D011720:Pyrazoles; C033607:puerarin