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目的:针对溃疡性结肠炎(ulcerative colitis,UC)活动期病机组成“清肠化湿、调气和血、敛疡生肌”的清肠化湿方,通过体外实验,观察其对小鼠骨髓来源树突状细胞(dendritic cells,DC)抗原提呈功能的影响,探讨该方治疗UC的作用机制。方法:以核因子-κB基因圈套寡核苷酸(NF-κB decoy ODN)转染24 h后的DC生物学特性为对照,观察10 mg.L-1清肠化湿方孵育24h后DC(细胞密度1×106/mL)细胞的生物学特性的改变,确立清肠化湿方通过抑制NF-κB的表达影响DC的抗原提呈功能。实验分为空白组,ODN转染组,清肠化湿方组,脂多糖(LPS)组,ODN转染+LPS组,清肠化湿方+LPS组6组。流式细胞术检测各组DC表面标志抗原CD11c,共刺激分子CD40,MHCⅡ的表达,免疫荧光法检测各组NF-κB核转位情况。结果:清肠化湿方+LPS组与LPS组CD40、MHCⅡ比较,P<0.05;各组与LPS组NF-κB核转位情况相比较,P<0.05。说明清肠化湿方有效降低了DC表面抗原CD40、MHCⅡ共刺激分子的表达,抑制了NF-κB的活化入核。结论:清肠化湿方通过抑制NF-κB的表达,影响DC成熟与分化,下调抗原提呈功能,从而降低炎症反应,是其治疗UC的主要机制。
Objective: To observe the effect of Qingchang Huashi decoction on the pathogenesis of ulcerative colitis (UC) during the pathogenesis of "Qingchanghu dampness, qi and blood, To investigate the effect of dendritic cells (DCs) on mouse bone marrow-derived dendritic cells (DCs) antigen presenting function and to explore the mechanism of action of this prescription in treating UC. Methods: The biological characteristics of DCs transfected with NF-κB decoy ODN for 24 h were compared. DC (a Cell density 1 × 106 / mL) cells to establish the biological characteristics of Qingchang Huashi decoction by inhibiting the expression of NF-κB DC antigen-presenting function. The experiment was divided into 6 groups: blank group, ODN transfection group, Qingchang Huashi Decoction group, lipopolysaccharide (LPS) group, ODN transfection + LPS group, Qingchang Huashi decoction + LPS group. Flow cytometry was used to detect the expression of CD11c, costimulatory molecules CD40 and MHCⅡ in each group. The nuclear translocation of NF-κB in each group was detected by immunofluorescence. Results: Compared with CD40 and MHCⅡ in LPS group, the content of NF-κB nuclear translocation in each group was significantly lower than that in LPS group (P <0.05). The results showed that Xiaozeng Huatan decoction effectively reduced the expressions of CD40 and MHCⅡ costimulatory molecules of DCs, and inhibited the activation of NF-κB. Conclusion: Qingchang Huashi Decoction can reduce the inflammatory reaction by inhibiting the expression of NF-κB, affecting the maturation and differentiation of DCs and down-regulating antigen presenting function, which is the main mechanism of its treatment of UC.