Dan He, Du Wen-ting, Fan Xue-jiao
Department of Pharmacy and Medical Laboratory Science, Sichuan College of Traditional Chinese Medicine, Sichuan.
Zhongguo Zhong Xi Yi Jie He Za Zhi. 2011 Dec;31(12):1674-9.
To study the flavanoids extracted from onion on the blood-brain barrier (BBB) permeation, and their effects on primary cultured neuron cell proliferation and apoptosis of SD rats using ethanol reflux method.
The brain microvascular endothelial cells (BMVECs) were first successfully primary cultured. Then rats BMVECs and astrocytes (ACs) were co-cultured to establish the in vitro BBB model. The flavanoids were extracted from onion using ethanol reflux method. The model was verified by transmission electron microscopy (TEM) and trans-epithelial electric resistance (TEER). The flavanoids permeability was tested using high performance liquid chromatography (HPLC). Meanwhile, rat neuron cells were cultured and exposed to H2O2 and flavanoids. Their effects on the cell proliferation and apoptosis were observed using MTT assay. The injury of neuron DNA was analyzed using single-cell gel electrophoresis (SCGE) and immunofluorescent assay.
The in vitro BBB model was successfully established by TEM and TEER. Results of HPLC proved flavanoids extracts could effectively permeate the BBB with the permeability of 60.58%. The extractive at 10 - 20 microg/mL showed obvious inhibition on the apoptosis of neuron cells induced by H2O2, and attenuated the injury of neuron DNA.
The flavanoids extracted from onion ethanol reflux method could effectively penetrate the BBB. They also showed obvious inhibition on the H2O2 induced neuron cell apoptosis and DNA injury.
采用乙醇回流法研究从洋葱中提取的黄酮类化合物对血脑屏障(BBB)的通透性,以及它们对原代培养的SD大鼠神经元细胞增殖和凋亡的影响。
首先成功原代培养脑微血管内皮细胞(BMVECs)。然后将大鼠BMVECs与星形胶质细胞(ACs)共培养以建立体外血脑屏障模型。采用乙醇回流法从洋葱中提取黄酮类化合物。通过透射电子显微镜(TEM)和跨上皮电阻(TEER)对模型进行验证。使用高效液相色谱法(HPLC)检测黄酮类化合物的通透性。同时,培养大鼠神经元细胞并使其暴露于过氧化氢和黄酮类化合物中。使用MTT法观察它们对细胞增殖和凋亡的影响。使用单细胞凝胶电泳(SCGE)和免疫荧光测定法分析神经元DNA的损伤。
通过TEM和TEER成功建立了体外血脑屏障模型。HPLC结果证明黄酮类化合物提取物能够有效透过血脑屏障,通透性为60.58%。10 - 20μg/mL的提取物对过氧化氢诱导的神经元细胞凋亡具有明显的抑制作用,并减轻了神经元DNA的损伤。
采用乙醇回流法从洋葱中提取的黄酮类化合物能够有效穿透血脑屏障。它们对过氧化氢诱导的神经元细胞凋亡和DNA损伤也表现出明显的抑制作用。