丹酚酸B对缺糖缺氧/复糖复氧的体外血脑屏障模型的影响

The influence of salvianolic acid B to blood brain-barrier with oxygen/glucose deprivation and reintroduction in vitro

目的探讨丹酚酸B对体外血脑屏障缺糖缺氧/复糖复氧的保护作用。方法利用人脑微血管内皮细胞与C6细胞非接触共培养建立体外血脑屏障模型并分为正常组、实验组和给药组。利用无糖Earle′s液代替培养液在93%N2、2%O2和5%CO2条件下孵育2h模拟缺糖缺氧环境。辣根过氧化物酶通透性实验检测血脑屏障通透性变化。免疫荧光和电子显微镜观察紧密连接变化情况。结果给药组对辣根过氧化物酶的通透性明显低于实验组(P<0.05);免疫荧光和超微结构图片显示正常的紧密连接连续和致密,而实验组紧密连结中断和疏松,给药组介于二者之间。结论丹酚酸B能直接保护缺糖缺氧/复糖复氧引起的血脑屏破坏和降低通透性,为丹酚酸B对血脑屏障的保护机制提供的研究基础。

Objective To explore the influence of salvianolic acid B（SalB）to blood brain-barrier（BBB）in vitro oxygen/glucose deprivation and reintroduction.Methods We used the human brain microvascular endothelial cells and C6 cells to build BBB model that in vitro,and the models was divided into normal groups,experiment groups,treatment groups.The Earle′s liquid and the condition of 93%N2,2%O2,5%CO2replaced the normal culture environment to simulate environment of oxygen/glucose deprivation and reintroduction.The changes of BBB permeability was detected by the horseradish peroxidase permeability test.The tight junctions（TJs）was detected by immunofluorescence and electron microscope.Results The permeability of treatment group was significantly lower than the experiment group（P〈0.05）;The TJs of normal group was continuous and density;the experiment group′s TJs was interruption and osteoporosis;and the observation of treatment group′s TJs was between of the two group above.Conclusion The SalB can directly protect the BBB during oxygen/glucose deprivation and reintroduction,and decrease the permeability.The investigation also provided the research foundation for the SalB to protect the BBB.