Jing Guang-Chan, Zhang Meng-Ren, Ji Chao, Zuo Ping-Ping, Liu Yu-Qin, Gu Bei
Department of Traditional Chinese Medicine, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, 100730, China.
Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, Beijing, 100005, China.
Chin J Integr Med. 2016 Nov;22(11):832-839. doi: 10.1007/s11655-016-2256-0. Epub 2016 May 25.
To determine the effect of medicated serum of Chinese herbal compound Naofucong (, NFC) on the microglia BV-2 cells viability and the transcription and expression of interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α) in microglia BV-2 cells to further explore the mechanisms underlying the protective effect of NFC on inflammatory process induced by high glucose.
The microglia BV-2 cells incubated in vitro were divided into different groups: the control group (25 mmol/L glucose), the model group (75 mmol/L glucose), high glucose media containing different dose medicated serum of NFC. After being cultured for 24 h, changes in IL-6 and TNF-α were measured by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay. The expression of surface marker CD11b of activated microglia was measured by confocal laser scanning microscope and Western blot. Nuclear factor-κB (NF-κB) p-p65 expression was analyzed by Western blot.
The model group obviously increased the expression of microglial surface marker CD11b and NF-κB p-p65 (all P<0.01), induced a signifificant up-regulation of release and the mRNA expression of IL-6 and TNF-α (P<0.01 or P<0.05). The medicated serum of NFC could obviously down-regulate the transcription and expression of surface marker CD11 b and NF-κB p-p65 (all P<0.01), and inhibit the mRNA and protein expression (P<0.01 or P<0.05) of inflflammatory cytokines, such as IL-6 and TNF-α, in microglia BV-2 cells cultured with high glucose for 24 h.
The inhibition of microglial activation and IL-6 and TNF-α expression induced by high glucose may at least partly explain NFC therapeutic effects on diabetes-associated cognitive decline diseases. Its underlying mechanism could probably be related to the inhibition of NFC on NF-κB phosphorylation.
观察中药复方脑复聪含药血清对小胶质细胞BV-2细胞活力及白细胞介素-6（IL-6）、肿瘤坏死因子α（TNF-α）转录和表达的影响，进一步探讨脑复聪对高糖诱导的炎症反应的保护机制。
体外培养的小胶质细胞BV-2分为不同组：对照组（25 mmol/L葡萄糖）、模型组（75 mmol/L葡萄糖）、含不同剂量脑复聪含药血清的高糖培养基组。培养24 h后，采用实时定量聚合酶链反应和酶联免疫吸附测定法检测IL-6和TNF-α的变化。采用共聚焦激光扫描显微镜和蛋白质印迹法检测活化小胶质细胞表面标志物CD11b的表达。采用蛋白质印迹法分析核因子κB（NF-κB）p-p65的表达。
模型组小胶质细胞表面标志物CD11b和NF-κB p-p65的表达明显增加（均P<0.01），IL-6和TNF-α的释放及mRNA表达显著上调（P<0.01或P<0.05）。脑复聪含药血清可明显下调表面标志物CD11b和NF-κB p-p65的转录和表达（均P<0.01），并抑制高糖培养24 h的小胶质细胞BV-2中IL-6和TNF-α等炎性细胞因子的mRNA和蛋白表达（P<0.01或P<0.05）。
抑制高糖诱导的小胶质细胞活化及IL-6和TNF-α表达可能至少部分解释脑复聪对糖尿病相关认知功能减退疾病的治疗作用。其潜在机制可能与脑复聪抑制NF-κB磷酸化有关。
