高糖暴露对小鼠脑微血管内皮细胞通透性的影响  

Effects of high glucose exposure on permeability of brain microvascular endothelial cells

作　　者：娄丹[1] 沙毅杰 帅怡[1] 霍倩[1] 李晨[1] 肖萍[1] 陶功华[1] LOU Dan;SHA Yi-jie;SHUAI Yi;HUO Qian;LI Chen;XIAO Ping;TAO Gong-hua(Shanghai Municipal Center for Disease Control and Prevention,Shanghai 200336,Chirui)

机构地区：[1]上海市疾病预防控制中心,上海200336

出　　处：《上海预防医学》2020年第1期71-77,共7页Shanghai Journal of Preventive Medicine

基　　金：上海市卫生和计划生育委员会(201440619);上海市第四轮公共卫生三年行动计划“高端海外研修团队培养计划”(GWTD2015S03)。

摘　　要：【目的】探讨高糖暴露对脑微血管内皮细胞通透性的影响。【方法】选取小鼠脑微血管内皮细胞系(bEnd.3)作为研究模型,检测跨内皮细胞电阻值(TEER)、碱性磷酸酶(ALP)及γ-谷氨酰转肽酶(γ-GT)的活性,对细胞模型的血脑屏障特征进行鉴定;再通过细胞形态学、细胞活力、细胞内乳酸脱氢酶活性及ZO-1、Occludin基因相对表达量的变化评价高糖暴露对脑微血管内皮细胞通透性的影响。【结果】细胞的TEER值、ALP及γ-GT的活性随细胞培养时间的增加而逐渐升高;细胞形态学结果显示,高糖作用下细胞数目明显减少,贴壁不稳;细胞活力实验显示,经葡萄糖暴露处理后,受试细胞活力呈剂量和时间依赖性降低;随着葡萄糖暴露浓度的增高,细胞内乳酸脱氢酶活性逐渐增高,各剂量组差异均有统计学意义(P<0.05)。此外,对紧密连接蛋白ZO-1和Occludin的mRNA表达水平进行检测发现,高糖暴露可抑制ZO-1、Occludin基因的表达,且表达水平呈剂量依赖性降低。【结论】体外培养的bEnd.3细胞模型具有血脑屏障的特征,高糖暴露能够抑制紧密连接蛋白ZO-1和Occludin的表达,该结果可能与脑微血管内皮细胞通透性的改变有关。[Objective]To investigate the permeability of brain microvascular endothelial cells under the condition of high glucose exposure.[Methods]The bEnd.3 cell line was chosen to detect the value of trans-endothelial electrical resistance(TEER),the activity of alkaline phosphatase(ALP)andγ-glutamyl transferase(γ-GT).Hence,the characteristics of blood-brain barrier in cell model were identified.The permeability of brain microvascular endothelial cells on high glucose exposure was evaluated by cell morphology,cell viability,intracellular lactate dehydrogenase activity and relative expression of ZO-1 and Occludin genes.[Results]The value of TEER,the activity of ALP andγ-GT increased gradually with increasing incubation time.The observation of cell morphology showed that the number of cells decreased significantly under high glucose exposure,and the adherence was unstable.Cell viability decreased with higher concentration of glucose or longer exposure time under high glucose exposure.The activity of lactate dehydrogenase was also decreased,and there were significant differences among the dose groups(P<0.05).In addition,the expression levels of tight junction protein ZO-1 and Occludin were further detected.It was found that high glucose exposure inhibited the expression of ZO-1 and Occludin genes in a dose-dependent manner.[Conclusion]The bEnd.3 cell line has the characteristics of blood-brain barrier.High glucose exposure inhibited the expression of tight junction protein ZO-1 and Occludin.The results might be related to the change of the permeability in brain microvascular endothelial cells.

关 键 词：高糖 血脑屏障 bEnd.3细胞系 紧密连接 

分 类 号：R114[医药卫生—卫生毒理学]