|Budget Amount *help
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2001: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2000: ¥2,000,000 (Direct Cost: ¥2,000,000)
In fiscal 2000, to analyze the relationship between exercise load and peripheral blood neutrophilic apoptosis, we measured neutrophilic apoptosis rates, cytokine levels, and superoxide dismutase activity before and after ergometer exercise in patients with suspected ischemic heart disease, and published the results. In addition, to investigate the effects of exercise on healthy individuals, We examined neutrophilic apoptosis and measured cytokine and SOD levels before and after ergometer exercise loading in healthy volunteers.
In fiscal 2001, we investigated apoptosis rates using antibodies to CD16b and active caspase-3. Unlike the TUNEL method, this method has the advantage of being able to readily detect apoptosis using whole blood instead of isolated neutrophils. We found that, in healthy individuals who performed exercise that they felt was moderately hard, positive rates of active caspatase-3 decreased slightly below pre-exercise rates during exercise, reach a maximum after exercise, and decrease as the heart rate returned to normal. An association of these positive rates with the simultaneously measured SOD levels suggested the involvement of an oxidation-antioxidation mechanism. In addition, to analyze changes at the gene level, we isolated peripheral blood neutrophils from healthy individuals before, after, and during recovery from exercise, extracted RNA, and synthesized cDNA by reverse transcription. Using cDNA corresponding to 20 ng of total RNA, we performed real-time PCR on a Light Cycler, and measured the mRNA levels of the apoptosis-promoting BAX, apoptosis-inhibiting BCL-2, and G6PDH (as a housekeeping gene) genes. The expression level of BCL-2 mRNA was always extremely low. When the expression level of BAX mRNA was adjusted by dividing it by that of G6PDH mRNA, the adjusted BAX mRNA level was higher after than before exercise, and tended to rise further during recovery from exercise. The analysis of the expression level of A1 mRNA is in progress.