【摘要】 目的探讨2型糖尿病患者血清干预人脐静脉内皮细胞(ECV-304)对细胞骨架微丝及胞浆内游离钙浓度的影响。方法ECV-304传代后取对数生长期细胞分为正常对照组(N组)、健康人血清干预组(H组)和糖尿病患者血清干预组(MD组)。按照1×105/ml将细胞种植在Pe-tri皿中进行分组干预,其中健康人血清、糖尿病患者血清浓度分别占总体积的10%。采用Fluo-3/AM作为荧光指示剂在激光扫描共聚焦显微镜下观察细胞内游离钙浓度的变化,采用荧光探针标记的鬼笔环肽染色法观察细胞骨架微丝分布的差异。结果与N组比较MD组胞浆内荧光分布不均匀,胞浆内游离钙浓度显著升高,骨架微丝断裂,排列紊乱,少部分区域缺失。结论糖尿病患者血清干预正常的ECV-304可以造成内皮细胞的骨架结构改变,主要与细胞内游离钙浓度升高有关。更多还原

【Abstract】 Objective To explore the mechanisms of ECV-304 incubated by the serum of patients associated with diabetes.Methods ECV-304 were obtained by cultured conventionally and divided into N(control),H(cultured with the serum of healthy person) and MD(cultured with the serum of diabetes patients) groups.Model cells′ intracellular free calcium((Ca2+)i) and cytoskeleton were assayed by laser scanning confocal microscope(LSCM).Results ①There was a significant difference(P<0.001) regarding the intracellular calcium concentration((Ca2+)i) of ECV-304 between MD and H,N groups.② By the cytoskeleton,MD group was significantly different with the Group N.Conclusions The serum of diabetes patients could injury ECV-304 and cause model cells’ intracellular free calcium((Ca2+)i) and cytoskeleton change.更多还原