神经肽Y活化大鼠心肌细胞内Ca^(2+)及钙调素依赖的钙调神经磷酸酶途径  

Effect of NPY on Ca^(2+)/CaM-dependent calcineurin signaling pathway of cardiomyocytes in rat

作　　者：李晓云[1] 陈敏生[1] 黄少华[2] 董颀[3] 李荧辉[4] 张舒[5] 

机构地区：[1]广州医学院第二附属医院心内科,510260 [2]广州医学院第一附属医院心内科 [3]广州医学院生理教研室 [4]北京航天医学工程研究所 [5]西安第四军医大学航空生理系

出　　处：《现代临床医学生物工程学杂志》2006年第3期261-263,共3页Journal of Modern Clinical Medical Bioengineering

基　　金：广东省自然科学基金(4004529);国家自然科学基金(30440053)

摘　　要：目的观察神经肽Y(NPY)对心肌细胞内Ca2+及钙调素(CaM)依赖的钙调神经磷酸酶(CaN)信号传导途径的影响。方法以NPY刺激心肌细胞。用环胞素A(CsA)特异性抑制CaN活性。应用Western印迹测细胞内CaN-α蛋白表达。采用组织化学法测CaN酶活性。用Fluo3-AM荧光标记法观察心肌细胞胞质及核内钙离子浓度变化。结果加入100nmol/LNPY刺激后,心肌细胞CaN酶活力较对照组明显增高(P<0.05),10nmol/LNPY组和CsA组(100nmol/LNPY+5μg/mlCsA)CaN酶活性与对照组相比,差异无统计学意义。100nmol/LNPY组心肌细胞内CaN-α蛋白表达较对照组显著增高(P<0.05)。加入100nmol/LNPY在10min内,心肌细胞胞浆及核内钙含量与对照组相比,差异无统计学意义。经100nmol/LNPY刺激24h后,心肌细胞胞浆及核内钙含量明显高于对照组(P均<0.01)。结论NPY可活化心肌细胞内Ca2+及CaM-CaN途径。NPY刺激下产生的细胞内钙增加,可能是其活化CaN途径的始动环节。Objective To investigate the effect of neuropeptide Y （NPY） on Ca^2＋/CaM-dependent calcineurin （CaN） signaling pathway of cardiomyocytes in rats. Methods Cardiomyucytes of neonatal Wistar rats were cultured with NPY, Cyelosporine A （CsA） was used to inhibit the activity of CaN. Western blot and histochemistry were used to assess CaN expressiun and CaN activity, Fluorescent assay with Fluo3- AM was used to detect [ Ca^2＋ ] in plasm and nucleus of eardiomyoeytes. Results To compare with control group, the activity of CaN in N PY （ 100 nmol/L） group was enhanced remarkably （P 〈 0. 05 ）, There were no significant changes of CaN activity among NPY （ 10 nmol/L） group, CsA （ 100 nmol/L NPY ＋ 5 μg/ml CsA） group and control group. CaN-α expression was etevated considerably in cardiomyocytes stimulated by NPY（100 nmol/L） compared with control group（P 〈0. 05）. Within 10 min,there were no considerable changes of the [ Ca^2＋ ] in plasm and nucleus stimulated by NPY（ 100 nmol/L）. 24 h after incubation, the [ Ca^2＋ ] in plasma and nuclear in NPY（ 100 nmol/L） group was significantly elevated compared with control group（P 〈0. 01）, Conclusion NPY can activate the Ca^2＋/CaM-dependent calcineurin signaling pathway of cardiomyocytes, and the enhancement of [ Ca^2＋ ] induced by NPY may be the first step in it.

关 键 词：神经肽Y 钙调神经磷酸酶 细胞内钙浓度 大鼠 

分 类 号：R33[医药卫生—人体生理学]