摘要
研究小檗胺（Ber）对氯化钾（KC1）和哇巴因（Oua）诱导的家兔血管平滑肌细胞（VSMC）内游离钙（［Ca(2+)］i）的影响。培养的VSMC以荧光探针Fluo-3/AM负载，通过激光共聚焦显微镜检查法检测［Ca(2+)］i。细胞外钙2．0mmol·L(-1)时，VSMC静息时［Ca(2+)］i水平为（228±46）／FI（n＝26），且不被Ber30βmol·L(-1)所影响（P＞0．05）。但KCl60mmol.L(-1)升高的[Ca(2+)]i；可被Ber预处理所抑制（P＜0.01）。荧光强度达峰值的时间明显延长（P＜0．05或P＜0．01）。实验结果还显示，在标本中加入KC1或Oua，VSMC核内的荧光值不受影响。Ber的这些作用与Ver10μmoh.L(-1)的作用相似。本研究的结果显示，Ber可拮抗KC1或Oua升高［Ca(2+)］的作用，此抑制作用可能与Ber阻断电压依赖性钙通道及激活Na+－K+－ATPase的作用有关。
The effects of herbamine (Ber) on KCl- and ouabain-induced cytosolic-free calcium([Ca(2+)]i elevation were studied in cultured rabbit vascular smooth muscle cells (VSMC ). Cultured VSMC derived from rabbit aorta was loaded with fluorescence probe Fluo-3/AM and [ Ca2+ ], was measured by laser scanning confocal microscope (LSCM). In the presence of extracellular calcium 2. 0 mm0l .L(-1), the resting level of [Ca(2+)]i, was (228±46) (n = 26) and not affected by pretreatment with Ber (P> 0. 05). But pretreatment with Ber 30μmol. L(-l) decreased [Ca(2+)]i, elevations induced by KCl 60mmol.L(-1) (P＜ 0. 01 ) and ouabain 1μmol. .L(-1) (P＜ 0. 01 ). The time to peak of fluorescence change was also prolonged (P ＜0. 05 or P ＜0. 0l ). The results also showed that the changes of fluorescent intensity (FI) within nucleus of VSMC were not observed before and after addition of Ber. These effects of Ber On [Ca(2+)], elevation caused by KCl and ouabain was similar to those by pretreatment with Ver 10μmol.L(-l).The present results suggested that the antagonizing effects of Ber on [Ca(2+)], mobilization by KCl and ouabain might be involved in the inhibition of voltage-dependent calcium channels (VDCC) and activation of Na+ -K + -ATPase.
出处
《哈尔滨医科大学学报》
CAS
1999年第4期265-268,共4页
Journal of Harbin Medical University
关键词
小檗碱
Fluo-3/AM
钙
离子通道
血管平滑肌细胞
Berbamine
Fluo-3/AM
Calcium
Ion channels
Vascular smooth muscle cell
Confocal microscopy
