不同类型中药制剂逆转结直肠癌T细胞免疫抑制及其靶分子的比较研究  

Reversing effects and target molecules of different traditional Chinese medicines on T cell immunosuppressions caused by colorectal cancer cells

作　　者：崔澂[1] 张宇辉[1] 支国成[1] 傅占江[1] 张爱霞[1] 李文建[2] 王润田[2] 

机构地区：[1]白求恩军医学院训练部,石家庄050081 [2]河北医科大学基础医学院免疫教研室,石家庄050017

出　　处：《免疫学杂志》2011年第11期973-978,共6页Immunological Journal

基　　金：河北省科技支撑计划项目(092061111D)

摘　　要：目的比较研究不同类型抗瘤中药制剂对结直肠癌所致T细胞免疫抑制的影响,寻找能有效逆转结直肠癌T细胞免疫抑制的抗瘤中药制剂组方。方法分别制备三氧化二砷(As2O3)、川芎嗪(LHC)、黄芪(AMB)、氧化苦参碱(MOX)、猪苓多糖(PUPS)、青蒿琥酯(ART)等6种中药制剂作用后的小鼠结直肠癌Colon26细胞再培养上清;MTT法检测中药制剂作用后对Colon26所致T细胞转化抑制的影响,直接免疫荧光FCM法检测对IL-2Rα及CD3ε+ζ+表达抑制的影响;定量ELISA法测定上清中TGF-β1、VEGF、IL-4、IL-6、IL-10和PGE2含量;多元相关分析中药制剂逆转结直肠癌T细胞免疫抑制的相应靶分子。结果 AMB、PUPS及ART对T细胞转化抑制的下调程度较强(下调约50%),维持时间较长;ART对IL-2Rα表达抑制的下调作用最强(下调约50%),维持时间较长;AMB对CD3ε+ζ+表达抑制的下调作用最强,不但可完全消除表达抑制,还可进一步促进表达,维持时间亦较持久。对Colon26肿瘤细胞所致转化抑制及CD3ε+ζ+表达抑制的逆转,LHC的作用靶分子为TGF-β1和IL-10,其他5种中药制剂的靶分子均为TGF-β1。对IL-2Rα表达抑制的逆转,除AMB以外其他5种中药制剂的靶分子均为TGF-β1。结论通过阻碍肿瘤细胞分泌TGF-β1、IL-10等免疫抑制分子,以逆转肿瘤细胞产生的T细胞免疫抑制,应是中药制剂的新型抗瘤机制之一。Colorectal cancer cells exert immunosuppression via immunosuppressors to escape from immunological surveillance.Therefore,seeking for drugs that can reverse tumor immunosuppressions should be helpful to elevate clinical anti-tumor efficiencies greatly.Accordingly,this contrastive study was performed to analyze the reversing effects of different traditional Chinese medicines on T cell immunosuppressions caused by colorectal cancer cells.We collected supernatants of re-cultivated Colon26 cells（murine colorectal tumor cells） which were pre-treated by six traditional Chinese medicines（TCM） respectively,including As2O3,Ligustrazine Hydrochloride（LHC）,Astragalus Mongholicus Bge（AMB）,Matrine N-Oxide（MOX）,Polyporus Umbellatus Polysacchride（PUPS） and Artesunate（ART）.Suppressed transformation of T lymphocytes was measured by MTT,and suppressed expressions of IL-2Rα and CD3ε＋ζ＋ were measured by FCM of direct immunofluorescence labeling.The concentrations of six immunosuppressors（including TGF-β1,VEGF,IL-4,IL-6,IL-10 and PGE2） in supernatants were measured by quantitative ELISA.The results shown that the reversing effects of AMB,PUPS and ART on suppressed transformation were stronger（about 50%） and longer,as compared with other TCM,while ART demonstrated the strongest reversing effects on the suppressed IL-2Rα expression（about 50%） with longer duration time,and AMB showed the strongest reversing effects on the suppressed CD3ε＋ζ＋ expression,which reversed completely and then elevated significantly.The target molecules of LHC in down-regulated suppression of transformation and CD3ε＋ζ＋ expression were TGF-β1 and IL-10,and that of other five medicines were TGF-β1.The target molecules of five medicines except for AMB in down-regulated suppression of IL-2Rα expression were TGF-β1.In conclusion,reversing T cell immunosuppressions of tumor cells through decreasing immunosuppressors should be one of anti-tumor mechanisms of traditional Chinese medicines.

关 键 词：中药制剂 结直肠癌 Colon26 免疫抑制 T细胞 逆转 靶分子 

分 类 号：R285.5[医药卫生—中药学]