The Expression And Release Of Hsp60in6-OHDA Induced In Vivo And In Vitro Models Of Parkinson’s Disease

Objective To investigate the expression and release of Hsp60in6-OHDA induced rat model and PC12cell model of Parkinson’s disease (PD).Methods We first established6-OHDA induced rat model and PC12cell model of Parkinson’s disease (PD). In PD rat model, the number of TH positive cells in the midbrain was detected by immunohistochemistry. The TH and Hsp60protein levels were detected by western blot. Immunofluorescence showed the the expression and location of Hsp60in DA neurons, microglia and astrocytes. In the PC12cell model of PD, the expression of Hsp60in6-OHDA treated PC12cells and in the culture supernatant was detected by western blot. The expression and location of Hsp60in PC12cells was further detected by immunocytochemistry.Results:(1) In6-OHDA induced PD rat model, TH positive cell number and TH protein level after6-OHDA treatment were decreased gradually, beginning at3d and becoming significant at5d (P<0.05)(2) In6-OHDA induced PD rat model, the protein expression of Hsp60was decreased at3d after6-OHDA treatment, and to the lowest at1w (P<0.05), then rose back.(3) Immunofluorescence showed that some Hsp60was located outside of DA neurons after6-OHDA treatment.(4) Immunofluorescence showed that no expression of Hsp60was found in the rest microglia. But positive staining of Hsp60was found on the surface of activated microglia. (5) Immunofluorescence showed that the collocation of Hsp60and astrocytes could be detected, and the expression of Hsp60was increased in activated astrocytes (P<0.05).(6) The expression of Hsp60was decreased gradually in the6-OHDA treated PC12cell model of PD (P<0.05).(7) The expression of Hsp60was detected and increased in the culture supernatant of6-OHDA treated PC12cells (P<0.05).(8) By immunocytochemistry, the number of PC12cells was decreased after6-OHDA treatment, and the condensed or fragmented nucleus could be found. Besides, the expression of Hsp60was decrease gradually.Conclusion:In PD models, the expression change and release of Hsp60was detected, and Hsp60may take part in the progress of PD possibly by activating microglia.