脂肪酸结合蛋白7对脑缺血再灌注大鼠血脑屏障通透性的影响

Effect of fatty acid binding protein 7(FABP7)on the permeability of blood-brain barrier in rats with cerebral ischemia-reperfusion

目的探究脂肪酸结合蛋白7(FABP7)对脑缺血再灌注(I/R)大鼠血脑屏障(BBB)通透性的影响,及其涉及的机制。方法将大鼠脑微血管内皮细胞和星形胶质细胞共培养以建立BBB细胞模型。用氧葡萄糖剥夺再复氧(OGD/R)方式建立I/R损伤BBB模型;用FABP7重组蛋白(rh-FABP7)单独处理,或与S7155共处理此细胞模型。采用ENDOHM仪器、Western blotting法、流式细胞术分别测定细胞跨内皮电阻(TEER)、FABP7和紧密连接(TJ)以及基质金属蛋白酶(MMP)蛋白表达、细胞凋亡。建立大鼠脑I/R模型,腹腔注射rh-FABP7,采用神经功能评分法、湿重和干重法、伊文思蓝染色法分别测定大鼠神经功能、脑含水量、BBB通透性。结果过表达FABP7逆转OGD/R诱导的FABP7和TJ蛋白表达、及TEER的下调,以及MMP2/9表达和细胞凋亡的上调(P<0.05)。S7155处理促进FABP7对MMP2/9和TJ蛋白表达、TEER、以及细胞凋亡的作用(P<0.05)。过表达FABP7逆转I/R诱导的大鼠神经功能评分、脑含水量、BBB通透性上调及FABP7蛋白表达下调(P<0.05)。结论FABP7通过抑制MMP2/9,减轻脑I/R对BBB完整性的破坏。

Objective To investigate the effect of fatty acid binding protein 7(FABP7)on blood-brain barrier(BBB)permeability in cerebral ischemia/reperfusion(I/R)rats and the mechanisms involved.Methods BBB cell model was established by co-culture of rat brain microvascular endothelial cells and astrocytes.The BBB model of I/R injury was established by using oxygen-glucose deprivation and reoxygenation(OGD/R)method.This model was treated with the FABP7 recombinant protein(rh-FABP7)alone or together with S7155.ENDOHM instrument,Western blotting and flow cytometry were used to determine the transendothelial electrical resistance(TEER),FABP7,TJ and matrix metalloproteinase(MMP)protein levels,and apoptosis.The rat brain I/R model was established,and rh-FABP7 was injected intraperitoneally.The neurological function score,wet and dry weight method,and Evans blue staining were used to determine the neurological function,brain water content,and BBB permeability of rats.Results FABP7 overexpression reversed the OGD/R-induced down-regulation of TEER,and FABP7 and TJ protein expression,as well as the up-regulation of MMP2/9 expression and apoptosis(P<0.05).S7155 treatment promoted the effect of FABP7 on MMP2/9 and TJ protein levels,TEER,and apoptosis(P<0.05).Moreover,FABP7 overexpression reversed the I/R-induced increase of neural function score,brain water content,and BBB permeability and decrease of FABP7 protein level in rats(P<0.05).Conclusion FABP7 alleviated the damage of brain I/R to BBB integrity by inhibiting MMP2/9.