摘要
背景:树突状细胞在过敏性紫癜发病机制中可能具有重要的地位。已有报道黄芪在体内可以纠正过敏性紫癜患儿的Th1/Th2失衡,但目前关于黄芪在树突状细胞水平对过敏性紫癜病免疫功能调节的影响尚不清楚。目的:观察过敏性紫癜患儿外周血树突状细胞功能变化及黄芪对其的影响。设计、时间及地点:病例对照分析,于2005-10/2007-12在青岛大学医学院附属医院儿科研究所完成。材料:急性期过敏性紫癜患儿外周静脉血28份作为过敏性紫癜组,以门诊体检的健康同龄儿童外周静脉血18份作为正常对照组。黄芪水提剂由四川百利药业有限公司提供。方法:采用密度梯度离心法获取过敏性紫癜患儿及健康儿童外周血单个核细胞,获取的外周血单个核细胞体外经重组人粒-巨噬细胞集落刺激因子、重组人白细胞介素4、重组人肿瘤坏死因子α以诱生树突状细胞,并培养8d,其中过敏性紫癜组又分成2组:对照组和黄芪干预组。主要观察指标:ELISA法检测过敏性紫癜患儿及健康儿童血浆γ-干扰素和白细胞介素4水平及培养上清液中白细胞介素10,12,18水平,流式细胞仪检测树突状细胞表面CD83、CD86(B7-2)和CD80(B7-1)的表达率。结果:过敏性紫癜患儿急性期外周血中Th1型细胞因子γ-干扰素水平减少,Th2型细胞因子白细胞介素4水平增加,γ-干扰素/白细胞介素4比值明显降低。过敏性紫癜患儿外周血单个核细胞经诱导培养的树突状细胞表面CD86表达率较正常对照组明显升高,且两组CD86表达率与血浆白细胞介素4水平皆呈显著正相关,与γ-干扰素/白细胞介素4比值皆呈显著负相关;CD80表达率明显低于正常对照组,且两组CD80表达率与血浆γ-干扰素水平及γ-干扰素/白细胞介素4比值均呈显著正相关。过敏性紫癜患儿树突状细胞分泌白细胞介素12水平低于正常对照组(P<0.05),而白细胞介素10,18水平均高于正常对照组(P<0.05),两组树突状细胞分泌白细胞介素12水平与血浆γ-干扰素水平皆呈显著正相关(P<0.01),与γ-干扰素/白细胞介素4比值皆呈正相关(P<0.01,P<0.05);白细胞介素10分泌水平与其血浆白细胞介素4水平皆呈显著正相关(P均<0.01),与γ-干扰素/白细胞介素比值皆呈负相关(P<0.01,P<0.05);白细胞介素18分泌水平与其血浆白细胞介素4水平皆呈显著正相关(P均<0.01),与γ-干扰素/白细胞介素4比值皆呈负相关(P<0.01,P<0.05)。黄芪干预组树突状细胞表面CD83、CD86和CD80的表达率与对照组差异不显著,树突状细胞分泌白细胞介素12水平高于对照组(P<0.01),白细胞介素10,18水平均低于对照组(P<0.05,P<0.01)。结论:①过敏性紫癜患儿外周血树突状细胞存在明显功能紊乱,表现为CD86表达率升高、CD80降低,白细胞介素12分泌减少、白细胞介素10和白细胞介素18增加,上述变化与Th1/Th2失衡关系密切。②黄芪主要是通过改变过敏性紫癜患儿树突状细胞分泌细胞因子的水平来调节其功能。
BACKGROUND： Dendritic cells may be play a significant role in the pathogenesy of Henoch-Schonlein Purpura. It was reported that Astragalus Membranaceus could correct the imbalance of Th1/Th2 in children with Henoch-Schonlein Purpura, but, at present, the influence of Astragalus Membranaceus on immune functions in children with Henoch-Schonlein Purpura has been unknown from dendritic cells. OBJECTIVE： To study the changes in peripheral blood dendritic cell functions and intervention study of Astragalus Membranaceus in vitro on them in children with Henoch-Schonlein Purpura. DESIGN, TIME AND SETTING： A case control experiment was performed at the Pediatric Research institute of Affiliated Hospital of Medical College of Qingdao University between October 2005 and December 2007. MATERIALS： Totally 28 peripheral blood samples of Henoch-Schonlein Purpura children in acute phase was taken as Henoch-Schonlein Purpura group, and 18 normal children who received the healthy examination as normal control group. Astragalus Membranaceus was supplied by Sichuan Baili Pharmaceutical Co., Ltd. METHODS： Peripheral blood mononuclear cells of Henoch-Schonlein Purpura children and normal children were taken by density gradient centrifugation, and induced with recombinant human granulocyte macrophage colony stimulating factor, recombinant human interleukin-4 and recombinant human tumor necrosis factor- a to make them transform into dendritic cells, and cultured in vitro for 8 days. Peripheral blood mononuclear cells of Henoch-Schonlein Purpura children were divided in two groups： control group and Astragalus Membranaceus group. MAIN OUTCOME MEASURES： Levels of interfere-y, interleukin-4 in plasma and interleukin-10, interleukin-12, interleukin-18 in supernatant were determined by enzyme linked immunosorbent assay in children with Henoch-Schonlein Purpura and normal children. Expression of the co-stimulatory molecules CD83, CD86 （B7-2） and CD80 （B7-1） on peripheral blood dendritic cells were examined by flow cytometry. RESULTS： Interfere- γ levels and the ratio of interfere- γ / interleukin-4 in plasma of Henoch-Schonlein Purpura children were significantly lower than those of the controls, respectively; while interleukin-4 levels were remarkably higher than those of the controls. Henoch-Schonlein Purpura children had remarkably increased expression of the CD86 on peripheral blood dendritic cells compared with the normal control subjects, and both of them had positive correlation with plasma interleukin-4 levels, and negative correlation with the ratio of interfere- γ/interleukin-4. Meanwhile, CD80 expression rate was significantly decreased in the Henoch-Schonlein Purpura children compared with the normal control subjects. CD80 expression rates in Henoch-Schonlein Purpura children and the normal controls were both positively correlated with the ratio of interfere- γ/interleukin-4. The levels of interleukin-12 produced by dendritic cells of Henoch-Schonlein Purpura children were lower than those of the normal controls （P 〈 0.05）, while interleukin-10, interleukin-18 levels were both higher than those of the normal controls （P 〈 0.05）. Between Henoch-Schonlein Purpura Children and the normal controls, the levels of interleukin-12 excreted by dendritic cells were both positively correlated with plasma interfere- γ levels （P 〈 0.01）, and also with the ratio of interfere- γ/interleukin-4 （P 〈 0.01, P 〈 0.05）. Interleukin-10 levels both had remarkably positive correlation with plasma interleukin-4 levels （P 〈 0.01）, and negative correlation with the ratio of interfere-γ/interleukin-4 （P 〈 0.01, P 〈 0.05）. Interleukin-18 levels had the same change with IL-10, respectively （P 〈 0.01）, （P 〈 0.01, P 〈 0.05）. No significant difference in CD83, CD86 and CD80 expression on peripheral blood dendritic cells was found between Astragalus Membranaceus group and control group. Supernatant interleukin-12 levels in Astragalus Membranaceus group were higher than those in control group （P 〈 0.01）, while supernatant interleukin-10, interleukin-18 levels were both lower than those in control group （P 〈 0.05, P 〈 0.01 ）. CONCLUSION： There is a disordered function of peripheral blood dendritic cells in Henoch-Schonlein Purpura children： elevated CD86 expression, lessened CD80 expression; decreased interleukin-12 levels, increased interleukin-10 and interleukin-18 levels, which are closely correlated to imbalance of Th1/Th2. It is through changing levels of cytokines produced by dendritic cells that Astragalus Membranaceus regulates its functions in children with Henoch-Schonlein Purpura.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2009年第1期157-160,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
