目的:探讨脑通汤对脑微血管内皮细胞(BMECs)缺氧/复氧损伤的保护作用及血脑屏障通透性的影响。方法:体外培养大鼠原代BMECs,缺氧/复氧法模拟脑缺血再灌注损伤。免疫组化检测内皮屏障抗原(EBA)蛋白表达;实时荧光定量PCR法检测紧密连接蛋白-5(Claudin-5)m RNA、紧密连接蛋白(ZO-1)m RNA的表达。结果:免疫组化示,与正常细胞组比较,模型组EBA蛋白表达明显下降(P〈0.05);与模型组比较,脑通汤大、中剂量含药血清组EBA蛋白表达显著升高(P〈0.05);但正常血清组与模型组比较差异无显著性(P〉0.05)。实时荧光定量PCR法示,与正常细胞组比较,模型组Claudin-5、ZO-1 m RNA的表达明显下降(P〈0.05);与模型组比较,脑通汤不同剂量含药血清组干预后,Claudin-5、ZO-1 m RNA的表达量明显上调,其中脑通汤大剂量含药血清组显著上调(P〈0.05)。结论:脑通汤可以保护脑微血管内皮细胞缺氧/复氧损伤,其保护机制可能与其上调EBA蛋白及Claudin-5、ZO-1基因的表达密切相关。
Objective To explore the effect of Naotong soup and blood brain barria in BMECs after hypoxia/reoxygenation injury. Methods Primary BMECs of rat were cultured. EBA protein and Claudin-5, ZO-1 gene were detected. Results Immunohistochemistry showed that the EBA protein was significantly decreased in model group compared with normal cells group. However, EBA protein was significantly higher than model group in large, medium doses of Naotong soup medicinal serum groups (P 〈 0.05 ) and no significant difference with normal serum group (P 〉 0.05). The sequencing expressions of gene Claudin-5, ZO-1 was broadly consistent EBA protein in Real-time PCR. Conclusion Naotong soup can protect the BMECs during bypoxia/reoxygenation injury. It is closely related to upregulate the EBA protein and Claudin-5, ZO-1 gene.