睡眠剥夺对大鼠血脑屏障的影响

Influence of Sleep Deprivation on Blood-brain Barrier in Rats

目的探讨睡眠剥夺(SD)对成年雄性大鼠血脑屏障的影响。方法健康雄性Sprague-Dawley大鼠90只随机等分为SD组、SD恢复(SDR)组和对照(K)组。SD组、SDR组大鼠采用水平台法连续SD5d,SDR组在SD后再正常饲养2d。K组不采取措施。Evens Blue(EB)心脏灌注观察大脑染液渗漏情况,Western blotting检测皮质下组织zonula occludens-1(ZO-1)、Occluding、Claudin-5、Bax/BCL-2、P53、caspase-3含量,免疫荧光染色观察大脑皮质内皮细胞、神经元细胞及其凋亡情况。结果SD组多个脑叶广泛皮质可见EB染液渗出;SDR组相应部位也有染液渗出,但较轻微;K组未见染液渗出。P53、Caspase-3、Bax/BCL-2表达SD组最多,K组最少;ZO-1、Occludin、Claudin-5表达SD组最少,K组最多;SDR组均介于两者之间(F>39.915,P<0.001)。皮质中CD31、NeuN阳性细胞数SD组最少,K组最多;TUNEL阳性细胞SD组最多,K组最少;SDR组介于两者之间(F>142.056,P<0.001)。结论SD能导致大鼠血脑屏障通透性增加,紧密连接蛋白表达降低,神经元凋亡增加,皮质神经元和内皮细胞数目减少;睡眠恢复可部分缓解其对血脑屏障的影响。

Objective To investigate the influence of sleep deprivation (SD) on blood-brain barrier in adult male rats. Methods A total of 90 healthy male Sprague-Dawley rats were randomly divided into SD group,SD and recovery (SDR) group and control (K) group.SD group and SDR group were continuously deprived of sleep for five days by horizontal table,and then,SDR group were fed normally for two days after SD.K group accepted no intervention. The leakage of Evens Blue (EB) in brain was observed after EB perfusion.The expression of zonula occludens-1 (ZO-1),Occluding,Claudin-5,Bax/BCL-2,P53 and caspase-3 in the cortex was detected with Western blotting. The apoptosis of neurons and endothelial cells in cortex was observed with immunofluorescence staining. Results In SD group,EB was observed in multiple cerebral lobe and extensive cortex,and it was also observed in SDR group in a milder way,but not observed in K group.The expression of P53,Caspase-3,Bax/BCL-2 in the cerebral cortex was the most in SD group,and the least in K group;while the expression of ZO-1,Occluding and Claudin-5 was the least in SD group,and the most in K group,and it was in-between in SDR group (F > 39.915, P < 0.001).The CD31 and NeuN positive cells decreased in cortex were the least in SD group,and the most in K group,while the TUNEL positive cells were the most in SD group,and the least in K group,and the levels of CD31,NeuN and TUNEL positive cells were in-between in SDR group (F > 142.056,P < 0.001). Conclusion SD may lead to dysfunction of permeability of blood-brain barrier,while decrease expression of tight junction protein,and increase apoptosis of neurons in rats to reduce the neurons and endothelial cells in cerebral cortex. Sleep recovery may partly alleviate these impacts.