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目的观察双氢青蒿素(dihydroartemisinin,DHA)对脂多糖(lipopolysaccharide,LPS)诱导的BV-2小胶质细胞神经炎症反应的影响及其机制。方法 LPS诱导BV-2小胶质细胞活化建立炎症模型,用不同浓度DHA(0.5、1、2、4μmol/L)处理细胞,CCK-8检测细胞活性;选取1μmol/L DHA干预细胞,倒置相差显微镜下观察细胞形态的变化;RT-PCR检测iNOS、IL-1β、IL-6、TNF-α基因表达水平;ELISA检测培养基中IL-1β、IL-6、TNF-α的表达水平;Western blot检测NF-κB、IκBα及TLR4的蛋白表达。结果低剂量的DHA(<2μmol/L)对BV-2细胞活性无明显影响(P>0.05),DHA可抑制LPS引起的BV-2细胞形态变化,DHA抑制活化的BV-2细胞iNOS、IL-1β、IL-6、TNF-αmRNA的表达(P<0.01),减少IL-1β、IL-6、TNF-α炎症因子释放(P<0.01),明显降低TLR4的蛋白表达,减少细胞质内IκBα的表达,并抑制NF-κB向核内移位(P<0.01)。结论 DHA可能通过作用于TLR4/NF-κB通路,抑制LPS诱导的BV-2小胶质细胞NF-κB的激活,从而抑制炎症因子的产生,发挥抗炎作用。
Objective To investigate the effects of dihydroartemisinin (DHA) on neuroinflammation of BV-2 microglia induced by lipopolysaccharide (LPS) and its mechanism. Methods LPS induced the activation of BV-2 microglia cells to establish the model of inflammation. The cells were treated with different concentrations of DHA (0.5, 1, 2 and 4μmol / L), and the cell viability was detected by CCK-8. The changes of cell morphology were observed under microscope. The expression of iNOS, IL-1β, IL-6 and TNF-α were detected by RT-PCR. The levels of IL-1β, IL-6 and TNF- blot detection of NF-κB, IκBα and TLR4 protein expression. Results Low dose of DHA (<2μmol / L) had no effect on the activity of BV-2 cells (P> 0.05), DHA inhibited the morphological changes of BV-2 cells induced by LPS, DHA inhibited the activation of BV- 1β, IL-6 and TNF-αmRNA (P <0.01), and the release of IL-1β, IL-6 and TNF-αincreased significantly (P <0.01), and significantly decreased the protein expression of TLR4 and decreased the expression of IκBα , And inhibited NF-κB translocation (P <0.01). Conclusion DHA may inhibit the activation of NF-κB induced by LPS in TL-4 / NF-κB pathway and inhibit the production of inflammatory cytokines and exert anti-inflammatory effects.