高晶体-高胶体渗透压混合液对人脐静脉内皮细胞内钙离子浓度和线粒体膜电位的影响  

Effects of Hypertonic-hyperoncotic Solution on Intracellular Calcium Concentration and Mitochondrial Membrane Potential of Endothelial Cells under Hypoxia and Reoxygenation

作　　者：袁世荧[1] 张雪萍[1] 姚尚龙[1] 曾邦雄[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院麻醉学教研室,武汉430022

出　　处：《华中科技大学学报（医学版）》2008年第3期362-364,共3页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

摘　　要：目的探讨高晶体-高胶体渗透压混合液(HHS)对缺氧/氧再灌状态下人脐静脉内皮细胞内钙离子浓度([Ca2+]i)、线粒体膜电位和细胞存活率的影响。方法将人脐静脉内皮细胞分为A、B、C、D4组,A组细胞于常氧状态下(37℃,95%空气,5%CO2)培养24h;B组细胞经HHS作用15min后,在常氧状态下培养24h;D组细胞先经HHS处理15min,C、D组细胞在缺氧环境(37℃,95%N2,5%CO2)中培养8h,然后在常氧状态下培养16h。[Ca2+]i采用Fluo-2荧光分光光度计测定,线粒体膜电位采用罗丹明123(rhodamine123)流式细胞仪测量,细胞存活率则采用锥虫蓝染色法检测。结果常氧状态下培养的A、B组细胞[Ca2+]i、线粒体膜电位和细胞存活率间的差异无显著性意义(P>0.05)。与A组相比,缺氧/氧再灌状态下,C、D组细胞[Ca2+]i明显增加(P<0.05),线粒体膜电位和细胞存活率明显降低(均P<0.05)。而D组细胞[Ca2+]i较C组降低,线粒体膜电位和细胞存活率较C组明显增高。结论HHS可阻止缺氧/氧再灌引起的人脐静脉内皮细胞的钙超载,并减轻线粒体膜电位和细胞存活率的下降。Objective To investigate the effects of hypertonic-hyperoncotic solution （HHS） on intraeellular calcium concentration （[Ca^2＋ ]i ）, mitochondrial membrane potential and viability of human umbilical vein endothelial cells （HUVECs） under hypoxia and reoxygenation in vitro. Methods HUVECs were incubated in normoxic medium （37℃, 95% air, 5% CO2） without HHS （group A） or with HHS （group B） for 24 h. HUVECs were exposed to hypoxic medium （37℃, 95% N2, 5% CO2 ） for 8 h and then reoxygenated with normoxic medium containing HHS （group D） or in the absence of HHS （group C） for 16 h. [Ca^2＋ ]i was determined by using fluorescent spectrophotometer after cells were treated with Fluo-2. Mitochondrial membrane potential was measured by using rhodamine 123 and flow cytometry. Cell viability was analyzed by vital dye staining. Results No significant differences were found in [Ca^2＋ ]i, mitochondrial membrane potential and cell viability of HUVECs between groups A and B. Under hypoxia and reoxygenation, [Ca^z＋ ]i was significantly increased （P〈0.01）, and mitochondrial membrane potential and cell viability were remarkably decreased （P〈0.05）. However, as compared with those in group C, [Ca^2＋ ]i was declined, and mitochondrial membrane potential and cell viability were enhanced in group D （all P〈0. 05）. Conclusion HHS could reverse hypoxia/reoxygenation-induced intracellular calcium overload, and prevent the decrease of mitochondrial membrane potential and cell viability in HVECs.

关 键 词：高晶体-高胶体渗透压混合液 人脐静脉内皮细胞 缺氧/氧再灌 钙离子 线粒体 

分 类 号：R614[医药卫生—麻醉学]