Effect of calcitonin gene-related peptide pretreatment on anoxia/reoxygenation-induced injury of H9c2 cell cultured in high-glucose culture medium

Objective To investigate the effect of calcitonin gene-related peptide (CGRP)pretreatment on anoxia/rexogenation-induced injury of H9c2 cell cultured in high-glucose culture medium. Methods H9c2 cells was cultured in DMEM/F12 culture with low-glucose and 10% fetal bovine serum. For establishing anoxia-rexogenation(AR) injury model of cardiac muscle cells, rat cardiac myoblasts (H9c2 cells) were exposed to 3 hours of anoxia followed by 3 hours of reoxygenation. Cultured H9c2 cells were divided randomly into five groups(n=6): Normal glucose-cultured control group(group NC), which was cultured in low-glucose culture for 48 h; High glucose-cultured control group(group HC), which was cultured in high-glucose culture for 48 h; High glucose-cultured AR group(group HAR), which was exposed to 3 hours of anoxia followed by 3 hours of reoxygenation(AR) after cultured in high-glucose culture for 48 h; High glucose-cultured pretreatment group(group HCP), which was pretreatmented by CGRP(final concentration for 10-8 mol/L) for 30 min followed by AR; High glucose-cultured CGRP and CGRP8-37 pretreatment group(Group HCCP), which was pretreatmented by CGRP 8-37 by 10 min followed by CGRP (final concentration for 10-7 mol/L) pretreatment for 30 min and then suffer AR progress. Results Compared with group NC, all cells of other groups had significant lower SOD activity and higher levels of LDH and apoptosis ratio of H9c2 cell(P<0.01). Compared to group HC, cells of group HAR and HNAR had significant decreased SOD activity of the cells and higher released LDH in cultured supernatant and apoptosis ratio of H9c2 cell(P<0.01). Compared with group HAR, cells of group HCP had significantly high SOD activity and decreased levels of the LDH and apoptosis ratio of H9c2 cell(P<0.01); Compared with group HCP, cells of group HCCP had significant decreased SOD activity of the cells and higher released LDH in cultured supernatant and apoptosis ratio of H9c2 cell(P<0.01). Conclusions It is suggested that CGRP pretreatment has direct role for decreaseing the AR-induced cell injury cultured in higher glucose medium, which is mostly associated with the activation of CGRP receptor. Key words: Calcitonin gene-related peptide; High glucose; H9c2 cell; Anoxia-reoxygenation