目的观察神经肽Y(NPY)刺激对大鼠心肌细胞肌浆网(Sarcoplasmic Reticulum,SR)内钙含量的影响。方法用100nmol·L^-1NPY刺激Sprague-Dawley乳鼠心肌细胞24h,用荧光染料Fluo-4AM负载胞浆钙,并用咖啡因诱导的胞浆钙瞬变(Caffeine-induced Ca2+ transient,CCT)幅度来反映肌浆网内总钙负荷;用荧光染料Fluo-5NAM直接标记心肌细胞肌浆网内游离钙离子。所有钙影像均由LeicaSP2激光共聚焦显微镜记录。结果经100nmol·L^-1NPY刺激24h后,心肌细胞肌浆网内游离钙含量明显低于对照组(p〈0.01);咖啡因诱导下钙瞬变幅度也低于对照组。结论长时间的NPY刺激可导致肌浆网内钙含量减少。
Objective To investigate the effect of neuropeptide Y (NPY) on calcium content in sarcoplasmic reticulum (SR) of rat cardiomyocytes.Methods Cardiomyocytes of neonatal Sprague-Dawley rats were incubated with NPY (100 nmol/L) for 24 h. Fluorescent assay with Fluo-4AM were used to detect Ca2+ transients of cardiomyocytes and Fluo-5N AM were used to detect[Ca^2+] in SR through laser scanning confocal microscope. The SR Ca^2+ load was estimated by Caffeine-induced Ca^2+ transient(CCT).Results 24 hours after incubation, compared with control group, the concentration of free Ca^2 + in SR( [ Ca^2 + ] ISR)was significantly decreased (p 〈 0. 05 ), and the peak of CCT was attenuated.. Conclusions NPY lead to reduce the calcium content in sareoplasmic reticulum of rat eardiomyoeytes after a long period of stimulation.