Guo Yuan-Yuan, Zheng Qin, Wu Ling, Zhang Ming-Xia, Zhang Ke-Nan, Xiao Shuai, Li Wen-Jing, Yang Ming
Key Laboratory of Modern Preparation of Traditional Chinese Medicine, Ministry of Education, Jiangxi University of Traditional Chinese Medicine Nanchang 330004, China.
Zhongguo Zhong Yao Za Zhi. 2020 Jun;45(12):2924-2931. doi: 10.19540/j.cnki.cjcmm.20200217.402.
According to traditional Chinese medicine, "spleen transport" is closely related to the metabolism of substance and energy. Studies have shown that Alzheimer's disease(AD) is a disease related to glucose and lipid metabolism and energy metabolism. The traditional Chinese medicine Jiangpi Recipe can improve the learning ability and memory of AD animal model. Sijunzi Decoction originated from Taiping Huimin Hefang Prescription is the basic prescription for strengthening and nourishing the spleen, with the effects of nourishing Qi and strengthening the spleen. In this experiment, human brain microvascular endothelial cells(HBMEC) and Sijunzi Decoction water extract(0.25, 0.5, 1 mg·L(-1)) were pre-incubated for 2 h, and then Aβ_(25-35) oligomers(final concentration 40 μmol·L(-1)) was added for co-culture for 22 hours. The effect of Sijunzi Decoction on the activity of Aβ_(25-35) oligomer injured cells and the expression of related proteins were investigated. Q-TOF-LC-MS was used first for principal component analysis of Sijunzi Decoction water extract. Then MTT assay was used to investigate the effect of Sijunzi Decoction water extract on the proliferation of HBMEC cells. Real-time fluorescence quantitative PCR(RT-qPCR) was employed to detect the mRNA expression of GLUT1, RAGE, and LRP1. The expression of Aβ-related proteins across blood-brain barrier(RAGE, LRP1) was detected by Western blot. The results showed that 40 μmol·L~(-1) Aβ_(25-35) oligomers could induce endothelial cell damage, reduce cell survival, increase expression of RAGE mRNA and RAGE protein, and reduce expression of GLUT1 mRNA, LRP1 mRNA, and LRP1 protein. Sijunzi Decoction water extract could reduce the Aβ_(25-35) oligomer-induced cytotoxicity of HBMEC, decrease the expression of RAGE mRNA and RAGE protein, and increase the expression of GLUT1 mRNA, LRP1 mRNA and LRP1 protein. The results indicated that Sijunzi Decoction could reduce the injury of HBMEC cells induced by Aβ_(25-35) oligomer, and regulate the transport-related proteins GLUT1, RAGE and LRP1, which might be the mechanism of regulating Aβ_(25-35) transport across the blood-brain barrier.
根据中医理论,“脾运化”与物质和能量代谢密切相关。研究表明,阿尔茨海默病(AD)是一种与糖脂代谢及能量代谢相关的疾病。中药降脾方可以改善AD动物模型的学习能力和记忆力。源自《太平惠民和剂局方》的四君子汤是健脾益气的基础方剂。在本实验中,将人脑微血管内皮细胞(HBMEC)与四君子汤水提取物(0.25、0.5、1 mg·L⁻¹)预孵育2小时,然后加入Aβ₂₅₋₃₅寡聚体(终浓度40 μmol·L⁻¹)共培养22小时。研究四君子汤对Aβ₂₅₋₃₅寡聚体损伤细胞活性及相关蛋白表达的影响。首先采用Q-TOF-LC-MS对四君子汤水提取物进行主成分分析。然后用MTT法检测四君子汤水提取物对HBMEC细胞增殖的影响。采用实时荧光定量PCR(RT-qPCR)检测GLUT1、RAGE和LRP1的mRNA表达。通过蛋白质印迹法检测血脑屏障相关Aβ蛋白(RAGE、LRP1)的表达。结果显示,40 μmol·L⁻¹ Aβ₂₅₋₃₅寡聚体可诱导内皮细胞损伤,降低细胞存活率,增加RAGE mRNA和RAGE蛋白表达,降低GLUT1 mRNA、LRP1 mRNA和LRP1蛋白表达。四君子汤水提取物可降低Aβ₂₅₋₃₅寡聚体诱导的HBMEC细胞毒性,降低RAGE mRNA和RAGE蛋白表达,增加GLUT1 mRNA、LRP1 mRNA和LRP1蛋白表达。结果表明,四君子汤可减轻Aβ₂₅₋₃₅寡聚体诱导的HBMEC细胞损伤,并调节转运相关蛋白GLUT1、RAGE和LRP1,这可能是其调节Aβ₂₅₋₃₅跨血脑屏障转运的机制。