Ye-Hao Zhang, Lan Miao, Peng Zhang, Guang-Yu Liu, Jian-Xun Liu
Beijing Key Laboratory of Pharmacology of Chinese Materia Region, Institute of Basic Medical Sciences,Xiyuan Hospital, China Academy of Chinese Medical Sciences Beijing 100091, China.
Zhongguo Zhong Yao Za Zhi. 2020 Oct;45(19):4686-4691. doi: 10.19540/j.cnki.cjcmm.20200511.401.
In this study, the oxygen-glucose deprivation(OGD) model in the human brain microvascular endothelial cell(HBMEC) was used to simulate the ischemic neuronal damage and observe the inflammatory response, explore the possible mechanisms for treating cerebral ischemia/reperfusion and improving memory impairment from the view point of inhibiting inflammatory response, which is of great reference significance for related Chinese medicine treatment of ischemic diseases. HBMECs were given with drugs at the same time of OGD injury, and reoxygenated for 2 h after 4 h treatment. Cell supernatant was then collected, and the inflammatory factors in cell supernatant were detected. Immunofluorescence assay was used to detect HBMECs morphology and expression of p-nuclear factor kappa-light-chain-enhancer of activated B(p-NF-κB); Western blot was used to detect expression changes of Toll like receptor 4(TLR4), myeloid differentiation primary response 88(MYD88) and p-NF-κB. The results showed that, after OGD modeling, the levels of interleukin 6(IL-6), IL-1α, IL-1β and tumor necrosis factor-α(TNF-α) were significantly increased; baicalin protected HBMEC, inhibited intranuclear transcription of p-NF-κB, significantly decreased HBMEC release of inflammatory factors caused by OGD injury, and inhibited the expression of TLR4, MYD88, and p-NF-κB. The studies suggested that baicalin had obvious protective effect on HBMECs damaged by OGD, and could inhibit inflammatory response. Its protection mechanism may be related to inhibiting TLR4 signaling pathways.
本研究采用人脑微血管内皮细胞(HBMEC)氧糖剥夺(OGD)模型模拟缺血性神经元损伤,观察炎症反应,从抑制炎症反应角度探讨治疗脑缺血/再灌注及改善记忆障碍的可能机制,对缺血性疾病的相关中医药治疗具有重要参考意义。在OGD损伤同时给予HBMECs药物,处理4 h后复氧2 h。然后收集细胞上清液,检测细胞上清液中的炎症因子。采用免疫荧光法检测HBMECs形态及磷酸化核因子κB(p-NF-κB)表达;采用蛋白质印迹法检测Toll样受体4(TLR4)、髓样分化因子88(MYD88)及p-NF-κB表达变化。结果显示,OGD建模后,白细胞介素6(IL-6)、IL-1α、IL-1β及肿瘤坏死因子-α(TNF-α)水平显著升高;黄芩苷对HBMEC具有保护作用,抑制p-NF-κB的核内转录,显著降低OGD损伤引起的HBMEC炎症因子释放,并抑制TLR4、MYD88及p-NF-κB表达。研究表明,黄芩苷对OGD损伤的HBMEC具有明显保护作用,并可抑制炎症反应。其保护机制可能与抑制TLR4信号通路有关。