Acta Pharmacologica Sinica (2010) 31: 219–226; doi: 10.1038/aps.2009.194

Department of Pharmacology, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China

Aim: To study the inducing effect of bicyclol on heat shock protein 27 (HSP27) and its role on anti-apoptosis in HepG2 cells intoxicated with D-galactosamine (D-GaIN).

Methods: HepG2 cells were treated with various concentrations of bicyclol and then subjected to D-GaIN intoxication.  Apoptosis was assayed by hoechst 33258 staining and flow cytometry analysis.  HSP27, cytochrome c, apoptosis inducing factor (AIF) and c-Jun N-terminal kinase (JNK) were assayed by Western blot.  Heat shock factor 1 (HSF1) was determined by electrophoretic mobility shift assay and the interactions of HSP27 with cytochrome c and AIF were detected by co-immunoprecipitation.    

Results: The results showed that bicyclol induced HSP27 protein and mRNA expression in HepG2 cells in both time- and dose-dependent manners (the maximal response: 1.23 fold increase at 100 µmol/L).  Bicyclol treatment stimulated HSF1 activation and increased the HSF1-HSE binding activity (the maximal response: 2.1 fold increase at 100 µmol/L).  This inducing effect of bicyclol on HSP27 and HSF1 was markedly blocked by quercetin.  Pretreatment of the cells with bicyclol markedly attenuated D-GaIN-induced apoptosis and the release of cytochrome c and AIF from mitochondria.  The induced HSP27 by bicyclol suppressed the activity of caspase-3 and the phosphorylation of JNK caused by D-GaIN in HepG2 cells.  All the above effect of bicyclol against D-GaIN-induced hepatocytes apoptosis were significantly reversed by quercetin.

Conclusion: HSP27 is involved in the anti-hepatocytes apoptosis of bicyclol, and this effect of bicyclol-induced HSP27 is mainly through inhibition of mitochondria and JNK apoptotic pathways.

Keywords: bicyclol; heat shock protein; mitochondria; c-Jun N-terminal kinase; apoptosis; HepG2 cells; cytochrome c; D-galactosamine