摘要
目的:体外研究青蒿琥酯(ART)逆转L929肿瘤细胞(L929)的免疫抑制作用。方法:获取经ART作用后再培养的L929培养上清和不经ART作用同步对照培养的L929培养上清,检测上清对小鼠脾细胞4项免疫功能的影响(包括NK杀伤和ConA诱导转化:MTT法;细胞表面CD4+和CD8+表达水平:流式细胞术法)和上清中4种免疫抑制分子的浓度(包括TGF-β1、PGE2、VEGF和IL-10:定量ELISA法)。分析青蒿琥酯逆转L929的免疫抑制作用与其下调L929免疫抑制分子分泌之间的关系。结果:同步对照培养的L929的上清,可稳定地检测到所测4种免疫抑制分子(TGF-β1和PGE2浓度最高)和对所测小鼠脾细胞4项免疫功能的显著抑制作用(C-S1与C-S2相比,P>0.05)。对ART作用后的L929:其第一次再培养上清(ART-S1)与相应对照上清(C-S1)相比,4种免疫抑制分子的浓度及对小鼠脾细胞NK杀伤、ConA诱导转化和CD4+CD8-表达的抑制作用显著降低(P<0.01);其第二次再培养上清(ART-S2)与ART-S1相比,除IL-10浓度轻度上升(P<0.05)和对小鼠脾细胞CD4-CD8+表达的抑制作用轻度降低(P<0.05)外,余无显著变化(P>0.05)。相关分析显示,ART逆转L929对小鼠脾细胞NK杀伤、ConA诱导转化及CD4+CD8-表达的抑制,与其下调L929分泌TGF-β1(r=0.971、r=0.984、r=0.990,P<0.05)、PGE2(r=0.960、r=0.981、r=0.982,P<0.05)及VEGF(r=0.970、r=0.992、r=0.982,P<0.05)显著正相关。ART逆转L929对小鼠脾细胞CD4-CD8+表达的抑制,可能与其下调L929其它免疫抑制分子分泌有关。结论:ART可通过下调L929免疫抑制分子分泌而逆转其免疫抑制作用,逆转肿瘤免疫抑制应是ART抗瘤新机制之一。
Objective： To investigate reversal effects of Artesunate （ART） on immunosuppression of L929 tumor cells （L929） in vitro. Methods：The supernatants of re-cultured L929 after treated with ART and the supernatants of synchronously cultured control L929 treated without ARTwere obtained. The effects of the supernatants on the immunofunctions of murine splenocytes including NK kiiling,ConA-induced transformation by MTY and the expression levels of CD4^＋ and CD8^＋ on ceil surface by FCM were：determined. The concentrations of four kinds of immunosuppressive molecules in the supernatants including TGF-β1 , PGE2, VEGF and IL-10 were detected by quantitative ELISA. The relationships between immunosuppression-reversal of ART and down-regulated on secretions of immunosuppressive molecules from L929 by ART were analyzed. Results： For the supernatants of synchronously cultured control L929, the four kinds of immunosuppressive molecules, in which the concentrations of TGF-β1 and PGE2were highest, and the significant inhibitory effect on the immunofunctions of murine splenocytes could be detected steadily （ C-S1 vs C-S2, P 〉 0. 05）. For the L929 after treated with ART： the supernatants from its first re-culture （ ART-S1 ） had lower concentrations of the four kinds of immunosuppressive molecules compared with the corresponding control supernatant （ C-S1 ）, and the inhibitory effect on the NK killing, ConA-induced transformation and CD4^＋ CD8^- expression of murine splenocytes significantly decreased （P 〈 0. 01 ）. The concentration of IL-10 was slightly increased （P 〈 0. 05 ）in the supernatants from second re-culture （ART-S2）compared with ART-S1, and the inhibitory effect on the CD4^- CD8^＋ expression of murine splenocytes slightly decreased （ P 〈 0. 05 ）. The others had not significant changes （ P 〉 0. 05 ）. The an.alysis of correlation indicated that there was the significantly positive correlations between ART reversal of the inhibitory effect by L929 on the NK killing, ConA-induced transformation and CD4^＋ CD8^- expression of murine splenocytes and ART down-regulation of the secretion of TGF-β1（r =0. 971, r =0.984, r =0.990,P〈 0.05）, PGE2（r =0.960, r =0.981, r =0.982,P〈0.05） and VEGF （ r = 0. 970, r = 0. 992, r = 0. 982 ,P 〈 0. 05 ） from L929. ART reversal of the inhibitory effect of L929 on the CD4^- CD8^＋ expression of murine splenocytes was perhaps related to ART down-regulation of secretion of immunosuppressive molecule from L929. Conclusion： ART could reverse the immunosuppression of L929 by down-regulating the secretion of immunosuppressive molecules from it, and the reversing tumor-induced immunosuppression should be one of the anti-tumor new mechanisms of ART.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2007年第11期985-988,共4页
Chinese Journal of Immunology
