Kim Byung-Wook, Koppula Sushruta, Park Shin-Young, Hwang Jin-Woo, Park Pyo-Jam, Lim Ji-Hong, Choi Dong-Kug
Department of Biotechnology, Konkuk University, Chungju, Korea.
BMC Complement Altern Med. 2014 Dec 16;14:502. doi: 10.1186/1472-6882-14-502.
A Saururus chinensis Baill (SC) has been used by Native Americans, early colonists and practitioners of Korean traditional medicine for treating several diseases including cancer, rheumatoid arthritis and edema. The objective of this study was to evaluate the effects of SC extract in lipopolysaccharide (LPS)-stimulated neuroinflammatory responses in BV-2 microglial cells.
The effects of SC on the LPS-induced neuroinflammatory responses in BV-2 microglial cells were assessed by Western blotting, RT-PCR and immunofluorescence labeling techniques. DPPH and alkyl radical scavenging assay was performed to evaluate the anti-oxidant effects. Comparisons between groups were analyzed using one-way analysis of variance followed by Dunnett's multiple comparisons test using GraphPad Prism V5.01 software.
Pre-treatment with SC extract (1, 5 and 10 μg/mL) significantly (p < 0.001 at 10 μg/mL) and concentration dependently inhibited LPS-induced production of nitric oxide (NO), inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2) and suppressed the inflammatory cytokine levels such as tumor necrosis factor-alpha and interleukin (IL)-6 in BV-2 microglial cells (p < 0.001 at 10 μg/mL). Further, SC suppressed the nuclear factor-kappa B (NF-κB) activation by blocking the degradation of IκB-α. SC also exhibited profound anti-oxidant effects by scavenging 1, 1-diphenyl-2-picrylhydrazyl (DPPH) (IC50: 0.055 mg/mL) and alkyl radicals (IC50: 0.349 mg/mL). High performance liquid chromatography finger printing analysis of SC revealed quercetin (QCT) as one of the major constituents compared with reference standard. QCT also inhibited the excessive release of NO, and inhibited the increased expressional levels of IL-6, iNOS and COX-2 in LPS-stimulated BV-2 cells.
Our results indicated that SC inhibited the LPS-stimulated neuroinflammatory responses in BV-2 microglia via regulation of NF-κB signaling. The antioxidant active constituents of SC might be partly involved in delivering such effects. Based on the traditional claims and our present results SC can be potentially used in treating inflammatory-mediated neurodegenerative diseases.
美洲原住民、早期殖民者和韩国传统医学从业者一直使用水龙(SC)治疗多种疾病,包括癌症、类风湿性关节炎和水肿。本研究的目的是评估水龙提取物对脂多糖(LPS)刺激的BV-2小胶质细胞神经炎症反应的影响。
通过蛋白质免疫印迹法、逆转录聚合酶链反应和免疫荧光标记技术评估水龙对LPS诱导的BV-2小胶质细胞神经炎症反应的影响。进行二苯基苦味酰基自由基(DPPH)和烷基自由基清除试验以评估抗氧化作用。使用GraphPad Prism V5.01软件,采用单因素方差分析,随后进行Dunnett多重比较检验分析组间差异。
用水龙提取物(1、5和10μg/mL)预处理可显著(10μg/mL时p<0.001)且浓度依赖性地抑制LPS诱导的一氧化氮(NO)、诱导型NO合酶(iNOS)、环氧化酶-2(COX-2)的产生,并抑制BV-2小胶质细胞中肿瘤坏死因子-α和白细胞介素(IL)-6等炎性细胞因子水平(10μg/mL时p<0.001)。此外,水龙通过阻断IκB-α的降解抑制核因子-κB(NF-κB)激活。水龙还通过清除1,1-二苯基-2-苦基肼(DPPH)(半数抑制浓度[IC50]:0.055mg/mL)和烷基自由基(IC50:0.349mg/mL)表现出显著的抗氧化作用。与参考标准品相比,水龙的高效液相色谱指纹图谱分析显示槲皮素(QCT)是主要成分之一。QCT还抑制NO的过度释放,并抑制LPS刺激的BV-2细胞中IL-6、iNOS和COX-2表达水平的升高。
我们的结果表明,水龙通过调节NF-κB信号传导抑制LPS刺激的BV-2小胶质细胞神经炎症反应。水龙的抗氧化活性成分可能部分参与发挥此类作用。基于传统说法和我们目前的结果,水龙可能潜在地用于治疗炎症介导的神经退行性疾病。