青藤碱对大鼠骨髓来源树突状细胞分化成熟的影响  

Sinomenine effects on differentiation and maturation of rat bone marrow-derived dendritic cells

作　　者：黄江波[1] 罗志刚[1] 高红强[2] 刘利[1] 何群君[1] 李建军[1] 言彩红[1] 龙向阳[1] 

机构地区：[1]南华大学附属第二医院,湖南省衡阳市421001 [2]湖南省儿童医院,湖南省长沙市410007

出　　处：《中国组织工程研究》2017年第21期3394-3399,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金面上项目(81273754)~~

摘　　要：背景:利用未成熟树突状细胞诱导供体特异性免疫耐受有可能成为防治器官移植排斥反应的重要途径。目的:探讨青藤碱对大鼠来源骨髓树突状细胞体外分化及成熟的影响。方法:取大鼠股骨、胫骨骨髓来源树突状细胞的前体细胞,经粒-巨噬细胞集落刺激因子及白细胞介素4作用诱导出未成熟树突状细胞,第7天加入脂多糖刺激成熟,在刺激未成熟树突状细胞成熟时,分为对照组和低、中、高不同剂量的青藤碱处理组,作用48 h后收获树突状细胞,倒置相差显微镜观察细胞形态学特征,流式细胞仪检测细胞表型CD80、RT1B的表达,ELISA检测培养上清白细胞介素12水平,混合淋巴细胞反应检测树突状细胞刺激同种异体T淋巴细胞活化的能力。结果与结论:(1)倒置显微镜下观察,对照组可见细胞符合成熟树突状细胞形态特性;青藤碱低剂量组可见绝大部分树突状细胞成熟;青藤碱中剂量组细胞部分悬浮,成熟度差;青藤碱高剂量组大部分细胞尚未成熟;(2)与对照组相比,青藤碱低、中、高剂量组CD80表达显著降低(P<0.05);与对照组相比,青藤碱中、高剂量组RT1B表达显著降低(P<0.05);(3)与对照组相比,青藤碱中、高剂量组培养上清中IL-12p70水平显著降低(P<0.01);(4)与对照组相比,青藤碱中、高剂量组刺激T淋巴细胞增殖的能力显著降低(P<0.05);(5)结果表明,青藤碱能抑制树突状细胞进一步成熟。BACKGROUND： It may be an important approach to avoiding organ transplant rejection by utilizing immature dendritic cells to induce donor-specific immunologic tolerance. OBJECTIVE： To study the effect of sinomenine on the differentiation and maturation of rat bone marrow-derived dendritic cells in vitro. METHODS： Bone marrow-derived dendritic cells were isolated from the rat femur and tibia, and immature dendritic cells were induced by granulocyte-macrophage colony stimulating factor and interleukin-4. On day 7, lipopolysaccharide was added and the cells were cultured to generate mature dendritic cells. Cells were divided into control group and low-, middle-and high-dose sinomenine treatment groups（SNL, SNM, SNH groups）. Forty hours later, dendritic cells were harvested, and cell morphology was observed by inverted phase contrast microscope. The expression of CD80 and RT1 B was detected by flow cytometry. ELISA was used to detect the expression of interleukin-12. The mixed lymphocyte reaction was used to detect the ability of dendritic cells to stimulate the activation of allogeneic T lymphocytes. RESULTS AND CONCLUSION：（1） Under the inverted microscope, the morphology of mature dendritic cells was observed in the control group; in the SNL group most dendritic cells were visible; in the SNM group, there were partially suspended cells with poor maturation; and in the SNH group, most of the cells were not mature.（2） The expression of CD80 in the control group was significantly lower than that in the SNL, SNM and SNH groups（P 0.05）, and the expression of RT1 B was significantly reduced in the SNM and SNH groups than the control group.（3） Compared with the control group, the level of IL-12p70 in the cell supernatant was significantly decreased in the SNM and SNH groups（P 0.01）.（4） The ability of dendritic cells to stimulate T lymphocyte proliferation in the SNM and SNH groups was significantly decreased compared with the control group（P 0.05）. To conclude, sinomenine can inhibit

关 键 词：树突细胞 免疫耐受 青风藤属 组织工程 干细胞 分化 青藤碱 树突状细胞 分化成熟 国家自然科学基金 

分 类 号：R394.2[医药卫生—医学遗传学]