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目的:探讨应用激光共聚焦扫描显微镜(LSCM)技术检测缺氧状态的人肺微血管内皮细胞(HPMVEC)内钙离子(Ca2+)浓度动态变化的价值。方法:HPMVEC常规培养,按观察时间点不同分为5个缺氧培养组(1h hyp组、2h hyp组、4h hyp组、6h hyp组和8h hyp组)以及1个对照组(0h con组)共6个组,每组设8个复孔,应用LSCM技术测定缺氧后HPMVEC内Ca2+浓度水平及随时间推移的变化。结果:LSCM技术显示HPMVEC内Ca2+的荧光强度1h hyp组与0h con组比较、2h hyp组与1h hyp组比较、4h hyp组与2h hyp组比较、6h hyp组与4h hyp组比较、8h hyp组与6h hyp组比较有显著差异(P<0.05)。线性回归分析结果显示Ca2+荧光强度与缺氧时间成正相关(r=0.969,P<0.01)。结论:HPMVEC内Ca2+浓度随缺氧时间增长而增高;LSCM在动态检测缺氧状态下HPMVEC内的Ca2+浓度变化中具有明显优势。
Objective: To investigate the value of confocal laser scanning microscopy (LSCM) in detecting the dynamic changes of calcium ion (Ca2 +) concentration in hypoxic human pulmonary microvascular endothelial cells (HPMVEC). Methods: HPMVEC routine culture was divided into five hypoxic culture groups (1h hyp group, 2h hyp group, 4h hyp group, 6h hyp group and 8h hyp group) and 1 control group (0h con group) A total of 6 groups, each with 8 complex holes, the application of LSCM technology determination of hypoxia HPMVEC Ca2 + concentration levels and changes over time. Results: The fluorescence intensity of Ca2 + in HPMVEC was detected by LSCM technique. Compared with 0 h con group, 2 h hyp group and 1 h hyp group showed that compared with 2 h hyp group, Compared with 6h hyp group, there was significant difference (P <0.05). Linear regression analysis showed that Ca2 + fluorescence intensity was positively correlated with hypoxia time (r = 0.969, P <0.01). CONCLUSION: The concentration of Ca2 + in HPMVEC increases with the increase of hypoxia time. LSCM has obvious advantage in the change of Ca2 + concentration in HPMVEC during hypoxia.