神经肽Y对大鼠心肌细胞Ca2＋-钙调素依赖性蛋白激酶Ⅱ的影响  

Effect of neuropeptide Y on calcium/calmodulin-dependent protein kinase Ⅱ in rat cardiomyocytes

作　　者：林晓圳[1] 董颀[2] 张弋[3] 陈敏生[1] 

机构地区：[1]广州医学院第二附属医院心内科广州心血管疾病研究所,510260 [2]广州医学院生理教研室 [3]广州医学院第二附属医院急诊科,510260

出　　处：《中华生物医学工程杂志》2011年第1期25-28,共4页Chinese Journal of Biomedical Engineering

基　　金：国家自然科学基金（30440053、30570759）;广东省自然科学基金（04004529）

摘　　要：目的 探讨神经肽Y对大鼠心肌细胞ca2＋-钙调素依赖性蛋白激酶Ⅱ（CaMK Ⅱ）的影响.方法 原代培养SD乳鼠心肌细胞,分为对照组（n=6）和神经肽Y组（n=6）,神经肽Y组加入终浓度为100 nmol/L的神经肽Y,对照组不进行处理.处理15 min后采用同位素32P掺人法检测心肌细胞CaMK Ⅱ活性,免疫印迹法（Western blotting）检测磷酸化CaMK Ⅱ（p-CaMK Ⅱ）蛋白表达;处理24 h后激光共聚焦显微镜下记录心肌细胞静息状态下细胞核Ca2＋-浓度,实时定量PCR检测心肌细胞β-肌球蛋白重链（β-MHC）、心钠素mRNA表达.结果 神经肽Y刺激心肌细胞15 min后,神经肽Y组CaMK Ⅱ活性较对照组明显升高（336 094.80±10 744.61比273 301.50±16 737.99,P＜0.05）,P-CaMK Ⅱ蛋白表达明显增加.神经肽Y刺激心肌细胞24 h后,神经肽Y组心肌细胞胞核Ca2＋-浓度较对照组明显增加（226.87±17.37比84.04±6.50,P＜0.01）;实时定量PCR显示β-MHC、心钠素mRNA表达亦明显增加.结论 神经肽Y通过增加心肌细胞胞核Ca2＋浓度活化CaMK Ⅱ.CaMK Ⅱ可能在神经肽Y诱导心肌细胞肥大效应中起着重要作用.Objective To explore the effect of neuropeptide Y （NPY） on calcium/calmodulindependent protein kinase Ⅱ （CaMK Ⅱ ） in rat cardiomyocytes.Methods The cardiomyocytes from neonatal Sprague-Dawley （SD） rats were primarily cultured and divided into the control and NPY groups （n=6 each）.The NPY group was added with 100 nmol/L NPY （final concentration）, while the control group was left untreated.After 15 min, the activity of CaMK Ⅱ was detected by isotope 32P incorporation assay, and the expression of phospho - CaMK Ⅱ （p - CaMK Ⅱ ） protein by Western blotting.After 24 h, intranuclear concentration of Ca2＋ in resting cardiomyocytes was recorded under laser scanning confocal microscope （LSCM）.In addition, real-time PCR was used to detect the expressions of P-myosin heavy chain （β-MHC）mRNA and atrial natriuretic factor （ANF） mRNA in cardiomyocytes.Results After NPY stimulation for 15 min, the NPY group showed significantly elevated activity of CaMK Ⅱ （336 094.80±0 744.61 vs 273 301.50±±16 737..99, P＜0.05） and increased expression of p-CaMK Ⅱ protein in cardiomyocytes, compared with the control group.After 24 h, the NYP group showed a significant increase in intranuclear Ca2＋ concentration compared wiith the control group （226.87±17.37 vs 84.04±6.50,P＜0.01）.Moreover, real-time PCR also showed signficant increase in expressions of β- MHC and ANF mRNA.Conclusion The CaMK Ⅱ is activated with the increase of intranuclear Ca2＋ concentration by NPY, which may play a major role in NYP-induced cardiomyocyte hypertrophy.

关 键 词：神经肽Y Ca2+-钙调素依赖性蛋白激酶Ⅱ 肌细胞 心脏 心肌病 肥厚性 大鼠 

分 类 号：R363[医药卫生—病理学]