Yang Huan, Cheng Xiao, Yang Ying-Lin, Wang Yue-Hua, Du Guan-Hua
Beijing Key Laboratory of Drug Target Identification and Drug Screening, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China.
Beijing Key Laboratory of Drug Target Identification and Drug Screening, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing; State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China.
Neural Regen Res. 2017 Nov;12(11):1860-1864. doi: 10.4103/1673-5374.219048.
Ramulus Cinnamomi (RC), a traditional Chinese herb, has been used to attenuate inflammatory responses. The purpose of this study was to investigate the effect of RC extract on lipopolysaccharide (LPS)-induced neuroinflammation in BV2 microglial cells and the underlying mechanisms involved. BV2 cells were incubated with normal medium (control group), LPS, LPS plus 30 μg/mL RC extract, or LPS plus 100 μg/mL RC extract. The BV2 cell morphology was observed under an optical microscope and cell viability was detected by MTT assay. Nitric oxide level in BV2 cells was detected using Griess regents, and the levels of interleukin-6, interleukin-1β, and tumor necrosis factor α in BV2 cells were determined by ELISA. The expression levels of cyclooxygenase-2, Toll-like receptor 4 and myeloid differentiation factor 88 proteins were detected by western blot assay. Compared with the LPS group, both 30 and 100 μg/mL RC extract had no significant effect on the viability of BV2 cells. The levels of nitric oxide, interleukin-6, interleukin-1β and tumor necrosis factor α in BV2 cells were all significantly increased after LPS induction, and the levels were significantly reversed after treatment with 30 and 100 μg/mL RC extract. Furthermore, RC extract significantly inhibited the protein expression levels of cyclooxygenase-2, Toll-like receptor 4 and myeloid differentiation factor 88 in LPS-induced BV2 cells. Our findings suggest that RC extract alleviates neuroinflammation by downregulating the TLR4/MyD88 signaling pathway.
桂枝是一种传统的中药材,已被用于减轻炎症反应。本研究的目的是探讨桂枝提取物对脂多糖(LPS)诱导的BV2小胶质细胞神经炎症的影响及其潜在机制。BV2细胞分别用正常培养基(对照组)、LPS、LPS加30μg/mL桂枝提取物或LPS加100μg/mL桂枝提取物孵育。在光学显微镜下观察BV2细胞形态,并用MTT法检测细胞活力。用Griess试剂检测BV2细胞中的一氧化氮水平,并用ELISA法测定BV2细胞中白细胞介素-6、白细胞介素-1β和肿瘤坏死因子α的水平。通过蛋白质印迹法检测环氧合酶-2、Toll样受体4和髓样分化因子88蛋白的表达水平。与LPS组相比,30和100μg/mL桂枝提取物对BV2细胞活力均无显著影响。LPS诱导后,BV2细胞中的一氧化氮、白细胞介素-6、白细胞介素-1β和肿瘤坏死因子α水平均显著升高,用30和100μg/mL桂枝提取物处理后,这些水平显著逆转。此外,桂枝提取物显著抑制LPS诱导的BV2细胞中环氧合酶-2、Toll样受体4和髓样分化因子88的蛋白表达水平。我们的研究结果表明,桂枝提取物通过下调TLR4/MyD88信号通路减轻神经炎症。