吸烟、硝酸甘油对动脉粥样硬化家兔血管平滑肌细胞钙浓度的影响

目的研究吸烟、硝酸甘油对动脉粥样硬化血管平滑肌细胞胞内游离钙浓度(Ca2+]i)的影响,探讨吸烟对动脉粥样硬化形成及硝酸甘油的生物效应的作用。方法复制家兔动脉粥样硬化模型,分离血管平滑肌细胞。Fluo-3/AM负载细胞,应用流式细胞仪检测血管平滑肌细胞Ca2+]i,激光扫描共聚焦显微系统测定单个血管平滑肌细胞内Ca2+的时空变化。结果各组动物主动脉壁均见到程度不同的动脉粥样硬化斑块形成。动脉粥样硬化家兔血管平滑肌细胞Ca2+]i明显升高(48.45±5.31,与生理盐水对照组38.09±2.57比较,P<0.01);吸烟能增加动脉粥样硬化家兔血管平滑肌细胞Ca2+]i(56.48±2.99,与单纯动脉粥样硬化组48.45±5.31比较,P<0.01);硝酸甘油则能明显降低动脉粥样硬化家兔血管平滑肌细胞Ca2+]i(41.91±3.16,与单纯动脉粥样硬化组48.45±5.31比较,P<0.05);吸烟能明显抑制硝酸甘油降低血管平滑肌细胞Ca2+]i的作用(47.99±5.10,与硝酸甘油组41.91±3.16比较,P<0.05)。结论吸烟能明显增加动脉粥样硬化血管平滑肌细胞Ca2+]i,硝酸甘油则能明显降低动脉粥样硬化血管平滑肌细胞Ca2+]i,吸烟能抑制硝酸甘油的生物学效应。

Effects of smoking and glonoine on intracellular Ca2+ in vascular smooth muscle cell in rabbits with atherosclerosis

Aim To study the effects of smoking and glonoine on intracellular free Ca2+ concentration(Ca2+]i) in vascular smooth muscle cell in rabbits with atherosclerosis,and to explore the effects of smoking on atherosclerosis and biologic action of glonoine.Methods An atherosclerosis in rabbits was produced.The vascular smooth muscle cells were isolated.The cells were loaded by Fluo-3/AM.Ca2+]i in vascular smooth muscle cell was measured by flow cytometer(FCM).The spatial distribution and the dynamic changes of Ca2+]i in single vascular smooth muscle cells were determined by laser scanning confocal microscopy(LSCM).Results Atherosclerosis plaques in arteriae aorta were observed and the degrees were different in various groups.Ca2+]i in vascular smooth muscle cells in rabbits with atherosclerosis markedly increased(48.45±5.31) vs that in saline control(38.09±2.57),P<0.01].Smoking evidently induced the elevation of Ca2+]i in vascular smooth muscle cells in rabbits with atherosclerosis(56.48±2.99) vs atherosclerosis group(48.45±5.31),P<0.01].Glonoine markedly inhibited the elevation of Ca2+]i in vascular smooth muscle cells in rabbits with atherosclerosis(41.91±3.16) vs atherosclerosis group(48.45±5.31),P<0.05].Smoking remarkably suppressed the effect of glonoine on the decrease ofCa2+]i (47.99±5.10) vs glonoine group(41.91±3.16),P<0.05].Conclusions Smoking could markedly increase Ca2+]i in vascular smooth muscle cells in rabbits with atherosclerosis.Glonoine markedly inhibited the elevation of Ca2+]i in vascular smooth muscle cells in rabbits with atherosclerosis.Smoking remarkably suppressed the biological effect of glonoine.