中文摘要：

目的建立子鼠肠上皮细胞（IEC）添加人工基底膜体外原代培养，模拟缺血缺氧损伤后胞内钙含量、膜脂流动性、IEC凋亡及采用钙通道阻断剂的影响。方法建立子鼠IEC分离与原代培养和缺血缺氧模拟环境，设立对照组（A组），缺氧组（B组），缺血组（C组），缺血缺氧组（D组）及加维拉珀米2mg／L的相应对照组。利用流式细胞仪（FCM）计算凋亡细胞率，激光共聚焦显微镜检测胞内钙含量；自旋标记-顺磁共振技术（ESR）检测膜脂流动性。结果B、C、D组IEC凋亡较A组明显增加（P〈0．05）；加用钙离子阻断剂后B1、C1、D1组较相应对照组减少（P〈0．05）。锹血缺氧损伤导致胞内钙超载，尤以D组最显著，其次为B组，C组；加用钙通道阻断剂后细胞内钙含量下降，以D1组最明显（P〈0．01），依次为B1组，c1组。损伤组膜蛋白构像改变、运动速度减慢，表现为强固定化与弱固定化组分谱线高度比值（hs／hw）明显增大及膜蛋白旋转相关时间（Tc）显著延长，D组最为明显（P〈0．01），其次为B组，C组；加用钙通道阻断剂后hs／hw和Tc有不同程度的恢复，与损伤组比较，差异有统计学意义（P〈0．05）。结论缺血缺氧损伤后，膜脂流动性发生变化及胞内钙超载和IEC凋亡一致；钙通道阻断剂可降低胞内钙含量，减轻膜蛋白的构像改变，减少细胞凋亡。

英文摘要：

Objective To investigate the relationship between the apoptosis and intracellular calcium amount and the change of cellular membrane fluidity, and the influence of calcium antagonist. Methods Primary culture of rat intestinal epithelial cells （IECs） with construction of artificial basal membrane and ischemia and anoxia environment were established. IECs were divided into 8 groups, namely group A （control group）, group B （ anoxia group）, group C （ ischemia group）, group D （ ischemia and anoxia group） group A1 （A ＋ verapamil 2 mg/L） ,group B1 （B ＋ verapamil 2 mg/L） ,group C1 （C ＋ verapamil 2 mg/L） ,group D1 （D ＋ verapamil 2 mg/L）. Apotosis rate （AR） was detected by FCM,intracellular calcium overload by LSCM,and cellular membrane fluidity by spin labeling-ESR spectra. Results The AR in groups B,C and D groups was significantly higher than in group A （P 〈0.05）. The AR in groups B1, C1 and D1 that were treated with calcium channel antagonist was lower than in groups B,C and D. Ischemia and anoxia injury caused intracellular calcium overload,most significant in group D,followed by group B and group C （P 〈 0.01 ）. Calcium channel antagonist could markedly decrease the amount of intracellular calcium （P 〈 0.05）. The hs/hw ratio in groups B, C and D was increased significantly as compared with group A, so did the rotational correlation time （.re）, most significant in group D （P 〈 0.0! ）, followed by groups B and C. Calcium channel antagonist could markedly decrease the hs/hw ratio and Te （ P 〈 0.05）. Conclusion Under the injury of anoxia and ischemia, the AR of IECs was increased, and calcium overload and cellular membrane fluidity markedly changes. Calcium channel antagonist could decrease AR and intracellular calcium.and imvrove the cellular membrane fluidity.