摘要
目的用体外血脑屏障(BBB)模型,研究高温下基质金属蛋白酶9(MMP-9)对BBB微血管内皮细胞紧密连接蛋白claudin-1的影响。方法体外培养大鼠脑微血管内皮细胞和星形胶质细胞,经荧光黄渗透性测定,成功建立体外BBB模型。随机分为常温组(37℃)、高温组(39℃)、常温+MMP-9组(MMP-9 100ng/L)、高温+MMP-9组,并在30、60和90min用通透系数(Pe)值观察通透性变化。用免疫组织化学法和Western blot法检测claudin-1表达。结果高温组较常温组Pe值增加(P<0.05);高温+MMP-9组较上述2组进一步增加(P<0.05)。相同时间点高温组claudin-1阳性细胞较常温组降低(P<0.05)。90min时,高温+MMP-9组claudin-1阳性细胞较高温组降低[(75.0±12.8)个/HP vs(97.0±6.1)个/HP,P<0.05]。高温组claudin-1表达较常温组降低(P<0.05),高温+MMP-9组较相同时间点高温组进一步降低(P<0.05)。结论高温可导致体外BBB模型claudin-1表达下降,BBB通透性增加。外源性添加MMP-9能进一步加剧该损伤,提示高温可通过MMP-9加重BBB破坏。
Objective To study the effect of hyperthermia-induced MMP-9on expression of claudin-1in in vitro BBB model.Methods BMEC and astrocytes were cultured.An in vitro BBB model was established with its permeability assayed by LY test.The BBB model was randomly divided into 37℃ group,39℃ group,37℃＋MMP-9group and 39℃＋MMP-9group.The permeability of BBB model was recorded at 30,60 and 90min respectively.Expression of claudin-1in BMEC was detected by Western blot with immunocytochemical staining.Results The permeability of BBB model was higher in 39℃ group than in 37℃ group（P〈0.05）and was significantly higher in39℃＋MMP-9group than in 37℃ group（P〈0.05）.The number of claudin-1positive BMEC was smaller in 39℃＋MMP-9group than in 37℃ group at the same time points（P〈0.05）and in 39℃＋MMP-9group than in 39℃ group at 90min（75.0±12.8/HP vs 97.0±6.1/HP,P〈0.05）.The expression level of claudin-1was lower in 39℃ group than in 37℃ group（P〈0.05）and was significantly higher in 39℃＋MMP-9group than in 37℃ group（P〈0.05）,which further increased with the increasing time.Conclusion Hyperthermia can downregulate the expression of claudin-1in in vitro BBB model and upregulate the permeability of in vitro BBB model.Adding MMP-9can further aggrevate the damage of in vitro BBB model,indicating that hyperthermia can aggrevate the damage of in vitro BBB model through MMP-9.
出处
《中华老年心脑血管病杂志》
CAS
2017年第9期979-983,共5页
Chinese Journal of Geriatric Heart,Brain and Vessel Diseases
基金
国家自然科学基金(81460188)
云南省科技计划项目(2011FB148
2013FB201
2013IC041)
云南省中青年学术和技术带头人后备人才培养(2012HB028)
云南省卫生厅卫生系统学科带头人培养计划(D-201235)
关键词
基质金属蛋白酶9
血脑屏障
高温
诱发
内皮细胞
星形细胞
matrix metalloproteinase 9
blood-brain barrier
hyperthermia
induced
endothelial cells
astrocytes
作者简介
王利民现在九江市第一人民医院神经内二科工作通信作者：孟强，E—mail：mq301@sina．com.cn
