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目的探讨HBe Ag对LPS诱导小鼠骨髓源性树突状细胞(DC)成熟的影响。方法体外诱导C57BL/6小鼠骨髓细胞分化成未成熟树突状细胞,经CD11c磁珠分选纯化后将DCs随机分为空白对照组、LPS刺激组、HBe Ag+LPS刺激组。流式检测DC表型变化,混合淋巴反应(MLR)检测DC促T淋巴细胞增殖能力,酶联免疫法(ELISA)检测细胞上清液中IL-12的分泌水平,Western blot检测p38磷酸化水平,并设置SB203580组为阳性对照探讨细胞IL-12分泌的可能调节机制。结果 LPS刺激未成熟DC引起细胞表面MHC-Ⅱ、CD86表达升高,刺激同种异体淋巴细胞增殖能力增强,IL-12分泌量增高。HBe Ag可抑制LPS促进DC表面MHC-Ⅱ、CD86表达升高和促淋巴细胞增殖能力增强的作用。LPS刺激DC可引起p38磷酸化水平升高,并呈时间依赖性;HBe Ag或SB203580预处理细胞再予LPS刺激,磷酸化p38表达和IL-12分泌较单纯LPS刺激组明显下降。结论 HBe Ag对LPS引起的树突状细胞的成熟有一定的抑制作用,且HBe Ag可能通过抑制p38MAPK信号通路下调LPS诱导的树突状细胞IL-12的产生。
Objective To investigate the effect of HBeAg on LPS-induced mouse bone marrow-derived dendritic cells (DCs) maturation. Methods The bone marrow cells of C57BL / 6 mice were induced to differentiate into immature dendritic cells in vitro. DCs were randomly divided into blank control group, LPS stimulation group and HBe Ag + LPS stimulation group by CD11c magnetic beads sorting. Flow cytometry was used to detect the phenotypic changes of DCs. MLR was used to detect the proliferation of DCs. The levels of IL-12 in the supernatants were detected by enzyme-linked immunosorbent assay (ELISA) and the level of p38 phosphorylation , And set SB203580 group as a positive control to explore the possible regulatory mechanism of IL-12 secretion. Results LPS stimulation of immature DC caused the expression of MHC-Ⅱ and CD86 on the cell surface to increase, stimulate the proliferation of allogeneic lymphocytes and increase the secretion of IL-12. HBeAg can inhibit LPS to promote DC surface expression of MHC-Ⅱ, CD86 increased and lymphocyte proliferation ability. LPS-stimulated DCs induced the increase of p38 phosphorylation in a time-dependent manner. The cells pretreated with HBe Ag or SB203580 were stimulated with LPS, the phosphorylation of p38 and the secretion of IL-12 decreased significantly compared with LPS alone. Conclusion HBe Ag can inhibit the maturation of dendritic cells induced by LPS, and HBe Ag down-regulates the production of dendritic cells IL-12 by inhibiting the p38 MAPK pathway.