布地奈德对沙丁胺醇抑制平滑肌内游离钙浓度升高快速影响

Rapid Influence of Budesonide on Inhibiting Effect of Raising Concentration of Intracellular Free Calcium in Smooth Muscle with Albuterol

目的观察布地奈德(Budesonide,BUD)对沙丁胺醇(Albuterol,ALB)抑制气道平滑肌细胞内游离钙(Ca2+[i])浓度升高的快速影响作用。方法取原代培养豚鼠气道平滑肌细胞,通过共聚焦显微镜下荧光强度检测,比较不同浓度BUD(1×10-6mol/L、1×10-7mol/L、1×10-8mol/L)与ALB联合作用10 min ALB抑制胞内Ca2+[i]释放和胞外Ca2+内流作用的快速影响,同时应用米非司酮(Mifepristone,RU486)及放线菌酮(Cycloheximide,CHX)作为阻断剂进行实验,排除上述反应中的基因组效应。结果组胺引起胞内Ca2+[i]释放实验中,阳性对照组、ALB对照组、1×10-6mol/L、1×10-7mol/L及1×10-8mol/L BUD联合ALB组荧光强度峰值分别为89.6±27.1、63.6±23.4、44.3±18.0、51.9±22.8及59.2±22.8;外源性Ca2+内流实验中,阳性对照组、ALB对照组、1×10-6mol/L、1×10-7mol/L及1×10-8mol/L BUD联合ALB组荧光强度峰值分别为77.3±23.1、56.4±18.7、40.1±14.9、47.0±20.8和53.7±23.5。此两组实验中,1×10-6mol/L和1×10-7mol/L BUD联合ALB组Ca2+[i]荧光强度峰值与ALB对照组比较差异具有统计学意义(P<0.01或P<0.05)。而在胞内Ca2+[i]释放及外源性Ca2+内流实验条件下,RU486组及CHX组Ca2+[i]荧光强度峰值与1×10-6mol/L BUD联合ALB组比较差异均无统计学意义(P>0.05)。结论哮喘急性发作时,联合应用BUD能够增强ALB减轻细胞收缩强度的作用,快速改善气道痉挛症状,且此作用不能被糖皮质激素受体拮抗剂和蛋白合成抑制剂所阻断。

Objective To study rapid influence of Budesonide （BUD） on inhibiting effect of raising concentration of intracellular free calcium in airway smooth muscle with Albuterol （ ALB）. Methods Rapid influences of albuterol on intracel- lular calcium release and extracellular calcium influx in primary cultured cells of airway smooth muscle affected with different concentrations of BUD （1×10-6mol/L, 1 x lO-7mol/L and 1 x 10-8tool/L） combined with salbutamol for 10 minutes were detected by the fluorescence intensity under confocal microscopy, while the Mifepristone （RU486） and Cycloheximide （CHX） groups were tested as the blocking agent to exclude the genomic effect. Results The peaks of fluorescence intensity in posi- tive control group, ALB group, 1 × 10 -6 mol/L BUD group, 1 × 10-7 mol/L BUD group and 1 x 10 -s mol/L BUD group were 89.6 ±27.1, 63.6± 23.4, 44.3 ± 18.0, 51.9 ± 22.8 and 59.2 ±22.8 respectively in histamine-induced intracellular Ca2＋ [ i] release experiment; the peaks of fluorescence intensity in positive control group, ALB group, 1 x 10-6mol/L BUD group, 1 × 10 -Tmol/L BUD group and 1 x 10 -Smol/L BUD group were 77.3 ± 23.1, 56.4 ± 18.7, 40.1 ± 14.9, 47.0 ±20.8 and 53.7 ± 23.5 respectively in extraeellular calcium influxing experiment. In the two experiments, the differences in peaks of flu- orescence intensity in 1 x 10 -6 mol/L BUD group and 1× 10 7 mol/L BUD group were statistically significant, compared with that of ALB group （ P 〈 0. O1 or P 〈 0.05 ）. The differences in peaks of fluorescence intensity in RU486 group and CHX group were not statistically significant, compared with those of 1 × 10-6 mol/L BUD group and ALB group in histamine-induced in- traeellular Ca2 ＋ [ i] release experiment and extracellular calcium influxing experiment （P 〉 0.05）. Conclusion During a- cute asthma attack, Salbutamol combined with Budesonide can enhance its role of reducing cell contraction of strength and rap- idly relieve the airway spasm symptoms, and glucocorticoid receptor antagonists and protein synthesis inhibitor can not block its effect.