Effect of rotenone on Caspaseª²3, Fas and Fasl and relationship with apoptosis in PC12 cells

    ZHAO Qianª²Lu,CHAI Xiª²Qing

    Graduate College of Hebei Medical University, Shijiazhuang 050017, China

    ¡¾Abstract¡¿ AIM: To investigate the effect of rotenone on Caspaseª²3, Fas and Fasl and relationship with apoptosis in PC12 cells.METHODS: PC12 cells were treated with different concentrations of rotenone and the effect of rotenone on the ratio of apoptosis was tested by flow cytometry. 1.0 ¦Ìmol/L rotenone was added into PC12 cells culture medium at different times and the protein expression of Caspaseª²3 and Fas/Fasl was also tested using FCM. The morphological changes of apoptosis were observed under transmitting electron microscopy (TEM). RESULTS: The ratio of apoptosis in control group was 0.0199¡À0.0020. In the trial groups treated with different doses of rotenone for 2 d, the ratio increased as 0.0305¡À0.0030, 0.0486¡À0.0073, 0.0313¡À0.0041 and 0.0285¡À0.0031, respectively, significantly different from that in control group (P<0.02). The 1.0 ¦Ìmol/L rotenone had marked effect (P<0.01). When rotenone concentrations exceeded 1.0 ¦Ìmol/L, the effects decreased in a doseª²dependent manner. FI of Caspaseª²3, Fas and Fasl in control group was 1.0¡À0.03, 1.0¡À0.02 and 1.0¡À0.04, respectively. After treatment with 1.0 ¦Ìmol/L rotenone for 3, 6, 12, 24 and 48 h, the expression of Caspaseª²3, Fas and Fasl was observed in each group. The expression of Fas/Fasl was most significant after rotenone treatment for 6 h (P<0.01) and the expression of Caspaseª²3 was most significant after rotenone treatment for 12 h(P<0.01). The morphological changes under TEM were shown as unsmooth membrane, decreased cellular protruding, small ratio of nucleus/cytoplasm, increased space in cytoplasm, increased gap of periphery in nucleus uncertainly, and increased poikilochromatin collected in edge. CONCLUSION: Rotenone may damage the PC12 cells by activation of Caspaseª²3 and Fas/ Fasl apoptosis pathways. ......