摘要
目的检测哮喘豚鼠气道平滑肌细胞(ASMCs)内钙释放通道功能的改变,探讨与支气管哮喘的关系,同时,寻找传代培养ASMCs的方法。方法以Flou-3/AM为细胞内钙离子示踪剂,观察ASMCs在工具药作用下细胞内钙离子浓度([Ca2+]i)的改变。结果①在ASMCs外无钙情况下,不同浓度ryanodine(5×10-5,10-4,2×10-4mol·L-1)作用于原代培养正常与哮喘ASMCs,[Ca2+]i迅速升高,哮喘组明显高于正常组(P<0·01)。10-4mol·L-1的组织胺(hista-mine)作用于原代培养的正常组与哮喘组ASMCs,[Ca2+]i升高无差异(P>0·05)。②传代培养哮喘ASMCs在10-4、2×10-4mol·L-1ryanodine作用下,[Ca2+]i迅速升高,与原代细胞比较无差异(P>0·05)。在浓度为5×10-5mol·L-1时,原代明显高于传代(P<0.01)。哮喘组传代ASMCs对10-4mol·L-1的histamine反应不明显。结论哮喘豚鼠ASMCs内钙释放通道(RyRs)功能升高,特定条件下,哮喘传代细胞仍然保持原代细胞内钙释放通道的特性。
Aim To investigate the functional change of intracellular Ca^2＋ channels of cultured asthmatic guinea-pig airway smooth muscle cells （ASMCs） and to comprehend its relationship with asthma. Methods [ Ca^2＋ ] i was measured in presence of different pharmaceutical products with Flou-3/AM as a fluorescent Ca^2＋ indicator. The cell photofluorogram was gathered by a camera interlined with an inverted fluorescence microscope. Results （1） In extracellular Ca2^＋ -free condition, different concentrations of ryanodine （5 × 10^-5, 10^-4,2 × 10^-4 mol · L^-1 ） could induce [ Ca^2＋ ] i increase in primary cultured ASMCs, but the increase of [ Ca^2＋ ]i in primary cultured ASMCs of asthmatic guinea-pig was much higher than those of control group （P 〈 0.01 ）. In subcultured ASMCs of asthmatic guinea-pig, although in the presence of 5 × 10^-5 mol · L^-1 ryanodine, the increase of [ Ca^2＋ ] i was not so high as in primary cultured ASMCs（P 〈0. 01 ）, in the presence of 10^-4,2 × 10^-4 mol · L^-1 ryanodine, the increase of [ Ca^2＋ ] ; in both primary cultured ASMCs of asthmatic guinea-pig and subcultured ASMCs of asthmatic guinea-pig was at the similar level （ P 〉 0.05 ）.（2） In extracellular Ca^2＋ -free condition, histamine （10^-4mol·L^-1） induced changes of [ Ca^2＋ ] i in primary cultured ASMCs of group was not significantly both asthmatic and control different （ P 〉 0. 05 ）. Conclusion Ryanodine receptor of asthmatic guinea-pig showed hypersensitivity. Under specified condition, the characteristics of ryanodine receptor still retains in subcuhured ASMCs of asthmatic guinea-pig.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2007年第1期47-51,共5页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No30470765)
