离体细胞气泡损伤模型的建立及评估

目的 建立离体细胞气泡损伤模型,用于潜水减压病等气泡损伤相关疾病的发病机制研究。方法 研制离体细胞气泡损伤装置,以原代培养大鼠肺微血管内皮细胞（pulmonary microvascular endothelial cells,PMVEC）为研究对象,通过检测细胞活力变化和凋亡细胞染色,观察细胞损伤与气泡直径和气泡触碰时间之间的量效关系,最终建立稳定的离体细胞气泡损伤模型。结果 离体细胞气泡触碰装置包括气泡生成注射器、细胞培养皿和打孔装置,通过改变气泡生成注射器末端直径大小可以获得不同直径大小的气泡。0.5 mm直径气泡触碰1、2、3、4 h 均未对PMVEC细胞产生明显损伤（P均>0.05）,1.0、1.5和2.0 mm直径气泡触碰1、2、3、4 h虽然对PMVEC造成损伤（P均＜0.01）,但是只有2.0 mm直径气泡触碰时间>3 h时PMVEC细胞损伤率才较大,触碰3h时细胞存活率为（84.27±1.35）%。结论 本研究建立了稳定的细胞气泡触碰损伤模型,可用于减压病等气泡损伤相关疾病的研究。就PMVEC细胞而言,直径为2 mm气泡触碰损伤3 h可作为最适的气泡损伤条件。

Establishment and evaluation of cell bubble contact injury model

Objective To establish a cell bubble injury model for the study of the pathogenesis of bubble related diseases such as decompression sickness. Methods The cell bubble contact device was designed, and the dose-effect relationship between cell damage and bubble diameter and contact time was observed by detecting the changes of cell vitality and death cell staining in primary cultured rat pulmonary microvascular endothelial cells (PMVEC). Finally, a stable cell bubble injury model was established. Results The cell bubble contacting device included a bubble generating syringe, a cell culture dish and a punching device. Bubbles with different diameters could be obtained by changing the diameter of the end of the bubble generating syringe. There was no significant damage to PMVEC after 1, 2, 3, and 4 h of 0.5 mm bubble contacting (all P >0.05), while bubbles with diameter of 1.0, 1.5, and 2.0 mm did cause significant damage to PMVEC cells after contacting 1, 2, 3, and 4 h (all P < 0.01). However, the cell damage rate of PMVEC cells was obvious only when the 2.0 mm diameter bubble was touched for more than 3 h. The cell viability was (84.27±1.35) % when the bubble with diameter 2.0 mm contacted the PMVEC for 3 h. Conclusion This study established a stable cell bubble contact injury model, which can be used for the study of bubble damage related diseases such as decompression sickness. In addition, for PMVEC, 3 h of bubble contact with a diameter of 2.0 mm was the optimal condition for bubble damage.