清肠化湿方对小鼠骨髓来源树突状细胞生物学特性的影响  

Effect of Qingchang Huashi Formula on the Biological Characteristics of Mouse Bone Marrow-derived Dendritic Cells

作　　者：谷静[1] 沈洪[1,2] 刘军楼[1,2] 欧阳俊[1] 朱磊[1,2] 

机构地区：[1]南京中医药大学第一临床医学院,南京210029 [2]江苏省中医院消化科,南京210029

出　　处：《中国实验方剂学杂志》2013年第13期200-204,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81072778);江苏省自然科学基金项目(BK2011078);江苏省中医药局课题项目(JD11002;JD11011);江苏高校优势学科建设工程项目(JD11002;JD11011)

摘　　要：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 impact of Qingchang Huashi formula（QHF） on mouse bone marrow-derived dendritic cell（DC） antigen-presenting function and explore the mechanism of action of the treatment of UC.Method： The DCs ftransfected by nuclear factor κB decoy oligonucleotides（NF-κB ODN） were as control group to observe the change the characters of the DC after incubation of QHF,and then to observe whether QHF can influence the function of DC antigen-presenting by inhibiting the expression of NF-κB.The experiment was divided into six groups,which is blank group,QHF group,ODN transfection group,QHF and lipopolysaccharides（LPS） group,ODN transfection and LPS group.Flow cytometry was used to detect the DC surface of CD11c,CD40,MHC II expression and immune fluorescence was used to detect the nuclear translocation of NF-κB of each group.Result： QHF could effective ly reduce the DC surface antigens CD40 and MHC II costimulatory molecule expression,and inhibit NF-κB activation into the nucleus.Conclusion： By inhibiting the expression of NF-κB,affecting maturation and differentiation of DC,reducing the antigen-presenting function,thereby reducing the inflammatory response are the main mechanism of QHF treating UC.

关 键 词：清肠化湿方 树突状细胞 溃疡性结肠炎 核因子-ΚB 

分 类 号：R285.5[医药卫生—中药学]