SKF96365和氯化镍对环匹阿尼酸诱导的大鼠PASMC[Ca-(2+)]_i升高的影响.pdf

重庆医学2015年4月第44卷第11期
1445
论著
doi:10.3969/j.issn.1671-8348.2015.11.002
SKF96365和氯化镍对环匹阿尼酸诱导的大鼠
PASMCICA2+]升高的影响
彭公永,胡锦兴,邹义敏,彭芳,周玉民,胡国平,赵祝香
(广州医科大学附属第一医院/广州呼吸疾病研究所/呼吸疾病国家重点实验室/
呼吸疾病国家临床医学研究中心,广东广州510120)
摘要]目的研究SKF96365和化镍(NiCI2)对环匹阿尼酸(CPA)诱导的大鼠远端肺动脉平滑肌细胞( PASMC)内游离
钙离子浓度([Ca2r])变化的影响。方法培养大鼠 PASMC,运用荧光显微镜和 Incyte细胞内鈣浓度检测系统观CPA
SKF6365和NiCL2对 PASMC[Ca2+1的影响。结果含5mol/L硝苯地平的无钙 Krebs溶液解育 PASMC,10gmol/LCPA
使 PASMCLCA2t短暂小幅升高,恢复细胞外Cat至2.5mmol/L后,10mol/LCPA使 PASMC[Ca2+];迅速显著升高;50
mOJ/LSKF96365和500pmol/ L Nic2均能明显抑制10gmol/LCPA引起的 PASMC[Ca2+],升高,但对高钟(60mmol/LKCI)
溶液引起的 PASMC Lca2+1升高无影响。结论CPA可致大鼠 PASMC Tca2は3升高,且能被SKF96365和NiC12阻断,提示
CPA可能诱发细胞外Ca+经钙池操纵性钙通道(SOCC)内流,SKF96365和NiCl2能逃择性抑制SOCC活性使经SOCC的Ca2
内流减少。
[关键词]环匹阿尼酸;游离钙离子浓度;肺动脉平滑肌细胞;硝苯地平;钙池操纵性钙通道;大鼠
[中图分类号]R543.2
文献标识码]A
文章编号]1671-8348(2015)11-1445-04
Effect of SKF96365 and Niclz on cyclopiazonic acid induced intracellular calcium
cation concentration increase in rat distal pulmonary arterial smooth muscle cells
Peng Gongyong, Hu /inzing, Zou Yimin, Peng Fang Zhou Yumin, Hu Gwoping, Zhao Zhuxiang
(National Clinical Research Center for Respiratory Disease, National Key Laboratory of Respiratory Disease
Guangzhou Institute of Respiratory Diseases, First Affiliated Hospital
Guang zhou Medical University, Guangzhou, Guangdong 510120, China
Abstract] Objective To study the effect of SKF96365 and Nicl2 on cyclopiazonic acid(CPA) induced intracellular calcium
cation concentration ([Ca*1 change in rat distal pulmonary arterial smooth muscle cells(PASMC). Methods The rat distal
PASMC were isolated and cultured. The effects of CPA, SKF96365 and Nicl on [Ca: in PASMC were tested by fluorescence
microscope and In Cyte Ca * J]; measurement system. Results PASMC were incubated with Ca-free Krebs solution containing 5
pmol/L nifedipine, 10 pmol L CPA caused a small transient increase in Ca]i after restoration of extracellular Cato 5
mmo: /L, 10 mol/L CPA caused marked increases in [Ca* J in PASM incubated with Krebs solution containing 5 mol/L nife
dipine. Both 50 gmol/L SKF96365 and 500 umol/I Nicl distinctly attenuated the increases in [Ca" caused by 10 pmol/L CPA
in PASMC. However, neither 50 umol/L SKF96365 nor 500 mol/L Nicl affccted the increases in [Ca] caused by 60 mmol/L
Kcl in PASMC. Conclusion CPA induced increases in [Ca"3, may related to Caf release from sarcoplasmic reticulum and the in
flux of Ca through store-operated Ca channels (OCC)in rat distal PASMC. Both SKF96365 and Nicl could selectively block
SOCC and attenuated the influx of Ca?" through SOCC in PASMC
Lkey words cyclopiazonic acid; intracellular calcium cation concentration; pulmonary arterial smooth muscle cells; nifedipine
store-operated Ca"channels; rat
中国40岁以上人群慢性阻塞性肺疾病(COPD)患病率高性Ca2-ATP酶阻断剂,能够激活平滑肌细胞膜钙池操纵性钙
达8.2%],多数人经缺氧性肺动脉高压发展为慢性肺源性通道( store- operated Ca2+ channels,SOCC),参与调节平滑肌
心脏病。急性缺氧性肺血管收缩反应和慢性缺氧性肺血管结细胞内游离Ca2t浓度( intracellular Ca2+ concentration
构重塑是缺氧性肺动脉高压发生的两大主要环节,而缺氧[ca2+1)。本研究通过原代培养大鼠远端 PASMC,选用
对远端肺动脉收缩和重塑的影响比近端肺动脉更明显?,表SOC拮抗剂SKF96365和氯化镍(NiCl2),观察其对CPA诱
明远端肺动脉是肺动脉高压发病的主要部位。Ca2在肺动脉导的 PASMCにCa2+],改变的影响,为进一步研究缺氧引起的
平滑肌细胞( pulmonary arterial smooth muscle cells, PASMC)肺动脉收缩和血管重塑机制打下基础
的缺氩性收缩反应和增殖过程中发摔关键作用-。环匹阿1材料与方法
尼酸( cyclopiazonic acid,CPA)是平滑肌肌浆上的一种特异1.1材料主要试剂:Ⅰ型胶原酶、NaCI、KCl、KH2PO4、M