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目的研究黄芩苷(BAI)对LPS刺激诱导的DC2.4细胞的增殖、成熟及抗原呈递功能的影响,为进一步揭示黄芩苷抗炎的免疫细胞分子机制提供基础。方法 (1)采用CCK8法测定黄芩苷对LPS刺激的DC2.4增殖的影响;(2)采用流式细胞术检测BAI对LPS刺激的DC2.4细胞成熟标志分子CD86及抗原呈递分子MHCⅡ的影响。结果 (1)CCK8实验结果显示,DC2.4细胞培养12至24h后增殖明显(P<0.05),24h达到高峰,其后增殖速度降低;加入LPS作用48h后,DC2.4细胞仍有明显增殖(P<0.05);单用BAI高、中浓度在12h、24h时明显抑制DC2.4细胞增殖(P<0.05),单用BAI低浓度在3个时间点对DC2.4细胞增殖无明显抑制作用;在LPS+BAI组中,高、中浓度在3个时点均可明显抑制DC2.4的增殖,低浓度的BAI使LPS诱导的细胞增殖水平接近对照组细胞;(2)流式细胞仪检测结果表明,LPS组的CD86和MHCⅡ分子表达显著增加(P<0.01),BAI组的CD86和MHCⅡ分子表达有增加趋势(P>0.05),而LPS+BAI组的CD86和MHCⅡ表达显著下降(P<0.05)。结论黄芩苷抗炎的免疫分子机制可能与其抑制病原体刺激未成熟树突状细胞的成熟及其抗原呈递分子MHCⅡ的过度表达相关。
Objective To study the effects of baicalin (BAI) on the proliferation, maturation and antigen presenting function of DC2.4 cells stimulated by LPS, and provide the basis for further revealing the molecular mechanism of baicalin anti-inflammatory immune cells. Methods (1) CCK8 assay was used to determine the effect of baicalin on LPS-stimulated DC2.4 proliferation; (2) Flow cytometry was used to detect the effect of BAI on the maturation marker CD86 and antigen presenting molecule MHC II of LPS-stimulated DC2.4 cells . Results (1) The results of CCK8 assay showed that DC2.4 cells proliferated significantly at 12 to 24 hours (P <0.05), peaked at 24 hours, and then proliferated at a slower rate. After adding LPS for 48 hours, DC2.4 cells proliferated significantly (P <0.05). The proliferation of DC2.4 cells was significantly inhibited by BAI at high and medium concentrations at 12h and 24h (P <0.05), while at low BAI concentration alone, proliferation of DC2.4 cells was not inhibited . In LPS + BAI group, the proliferation of DC2.4 was significantly inhibited at high and medium concentrations at 3 time points, and BAI at a low concentration approached the LPS-induced cells proliferation in the control group. (2) Flow cytometry The results of ELISA showed that the expressions of CD86 and MHC Ⅱ were significantly increased in LPS group (P <0.01), and the expressions of CD86 and MHC Ⅱ molecules in BAI group were increased (P> 0.05), while the expressions of CD86 and MHC Ⅱ in LPS + BAI group were significantly decreased (P <0.05). Conclusion The immune mechanism of anti-inflammatory baicalin might be related to inhibiting the maturation of immature dendritic cells stimulated by pathogens and the over-expression of antigen presenting molecule MHCⅡ.