中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2013年
3期
222-226
,共5页
卢玉振%梁桂宁%陈璐璐%侯亚义
盧玉振%樑桂寧%陳璐璐%侯亞義
로옥진%량계저%진로로%후아의
灵芝多糖%Th1细胞%STAT4%IFN-γ
靈芝多糖%Th1細胞%STAT4%IFN-γ
령지다당%Th1세포%STAT4%IFN-γ
Ganoderma lucidum polysaccharide (GLP)%Th1 cells%STAT4%IFN-γ
目的 探讨灵芝多糖(GLP)对外周血淋巴细胞免疫分群的影响及其作用机制.方法 取肿瘤患者和正常人的外周血,分离外周血单个核细胞(PBMC)后,用不同剂量的GLP(10 ng/ml、50ng/ml和100 ng/ml)刺激后,用流式细胞仪检测DC细胞表面分子(HLA-DR、CD83和CD11c)、Th1细胞、Th2细胞和NK(CD3-CD56+)细胞数；并进一步用免疫磁珠分选出正常人外周血CD4+ Th细胞后用不同浓度GLP刺激24h后,荧光实时定量Q-PCR检测Th1和Th2细胞因子的表达水平,Westernblot分析Th1分化相关的转录因子水平.结果 灵芝多糖可以在体外呈浓度依赖性增加外周血中Th1细胞亚群和DC共刺激分子的表达(P＜0.01),并且增加STAT4的表达和IL-12、IFN-γ和TNF-α的mRNA的表达水平(P＜0.01).结论 灵芝多糖可能通过增加Th细胞STAT4的表达水平,促进其向Th1细胞分化,并增加Th1的分泌细胞因子.
目的 探討靈芝多糖(GLP)對外週血淋巴細胞免疫分群的影響及其作用機製.方法 取腫瘤患者和正常人的外週血,分離外週血單箇覈細胞(PBMC)後,用不同劑量的GLP(10 ng/ml、50ng/ml和100 ng/ml)刺激後,用流式細胞儀檢測DC細胞錶麵分子(HLA-DR、CD83和CD11c)、Th1細胞、Th2細胞和NK(CD3-CD56+)細胞數；併進一步用免疫磁珠分選齣正常人外週血CD4+ Th細胞後用不同濃度GLP刺激24h後,熒光實時定量Q-PCR檢測Th1和Th2細胞因子的錶達水平,Westernblot分析Th1分化相關的轉錄因子水平.結果 靈芝多糖可以在體外呈濃度依賴性增加外週血中Th1細胞亞群和DC共刺激分子的錶達(P＜0.01),併且增加STAT4的錶達和IL-12、IFN-γ和TNF-α的mRNA的錶達水平(P＜0.01).結論 靈芝多糖可能通過增加Th細胞STAT4的錶達水平,促進其嚮Th1細胞分化,併增加Th1的分泌細胞因子.
목적 탐토령지다당(GLP)대외주혈림파세포면역분군적영향급기작용궤제.방법 취종류환자화정상인적외주혈,분리외주혈단개핵세포(PBMC)후,용불동제량적GLP(10 ng/ml、50ng/ml화100 ng/ml)자격후,용류식세포의검측DC세포표면분자(HLA-DR、CD83화CD11c)、Th1세포、Th2세포화NK(CD3-CD56+)세포수；병진일보용면역자주분선출정상인외주혈CD4+ Th세포후용불동농도GLP자격24h후,형광실시정량Q-PCR검측Th1화Th2세포인자적표체수평,Westernblot분석Th1분화상관적전록인자수평.결과 령지다당가이재체외정농도의뢰성증가외주혈중Th1세포아군화DC공자격분자적표체(P＜0.01),병차증가STAT4적표체화IL-12、IFN-γ화TNF-α적mRNA적표체수평(P＜0.01).결론 령지다당가능통과증가Th세포STAT4적표체수평,촉진기향Th1세포분화,병증가Th1적분비세포인자.
Objective To study the effects of Ganoderma lucidum polysaccharide (GLP) on PBMCs and the related immune mechanism.Methods PBMCs from cancer patients and healthy donors were isolated and treated with different doses of Ganoderma lucidum polysaccharides (10 ng/ml,50 ng/ml and 100 ng/ml).DC cell costimulatory molecules (HLA-DR,CD83,and CD11 c),Th1 (CD3 + CD8-IFN-γ+) cells,Th2 (CD3 + CD8-IL-4+) cells and NK (CD3-CD56+) were analyzed by FCM.Furthermore,The CD3+ CD4+ Th ceils were separated by immunomagnetic beads and stimulated with Ganoderma lucidum polysaccharides at different concentrations in culture.After 24 h,the cytokine expression levels of Th1 and Th2were detected by RT-PCR.The expressions of Th1 differentiation-related transcription factor,STAT4,were analyzed by Western blot.Results Ganoderma lucidum polysaccharides can significantly stimulate in vitro Thl cell differentiation (P＜0.01) in a dose depend manner.It correlates with an increased expression of STAT4 and the elevated mRNA expression levels of Th1 cytokineincluding IL-12,IFN-γ and TNF-α (P＜0.01).Conclusion Ganoderma lucidum polysaccharides may promote Th1 differentiation and increase the secretion of Th1 cvtokines through the upregulation of STAT4.
