抗肿瘤中药制剂逆转结直肠癌NK细胞免疫抑制的靶分子探讨  

A study on the target molecules for anti-tumor traditional Chinese medicines to reverse NK cell immunosuppressions of colorectal cancer cells

作　　者：崔澂[1] 王润田[2] 胡建军[1] 王智华[2] 张征峥[2] 邓郁青[2] 

机构地区：[1]第四军医大学白求恩军医学院训练部,河北省石家庄市050081 [2]河北医科大学基础医学院免疫室

出　　处：《江苏医药》2011年第5期509-512,共4页Jiangsu Medical Journal

基　　金：河北省科技支撑计划项目(092061111D)

摘　　要：目的探讨抗瘤中药制剂逆转结直肠癌所致自然杀伤(NK)细胞免疫抑制的作用靶分子机制。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)6种中药制剂作用后的小鼠结直肠癌细胞Colon26再培养上清液,MTT法检测中药制剂对Colon26所致小鼠脾细胞NK杀伤抑制的影响,流式细胞仪直接免疫荧光法检测对小鼠脾细胞CD3ε-ζ+表达抑制的影响;ELISA测定上清液中转化生长因子(TGF)β1、血管内皮生长因子(VEGF)、白细胞介素(IL)4、IL-6、IL-10和前列腺素(PG)E2含量;多元相关分析中药制剂逆转结直肠癌NK细胞免疫抑制的相应靶分子。结果 AMB对NK杀伤抑制的逆转作用最强;LHC对CD3ε-ζ+表达抑制的逆转作用最强。对NK杀伤抑制的逆转,As2O3、AMB、PUPS和ART的靶分子为TGF-β1,LHC的靶分子为TGF-β1和IL-10;对CD3ε-ζ+表达抑制的逆转,AMB和MOX的靶分子为IL-10。结论通过抑制免疫抑制分子分泌逆转肿瘤细胞产生的NK免疫抑制,应是中药制剂抗瘤效应机制之一。临床可考虑联合使用不同类型抗瘤药物组方调整患者NK免疫抑制及免疫抑制分子分泌的状态。Objective To study the target molecules for different anti-tumor traditional Chinese medicines to reverse NK cell immunosuppressions of colorectal cancer cells.Methods Re-cultured supernatants of Colon26（murine colorectal tumor cells） pre-treated respectively by six different traditional Chinese medicines of As2O3,ligustrazine hydrochloride（LHC）,astragalus mongholicus bge（AMB）,matrine N-oxide（MOX）,polyporus umbellatus polysacchride（PUPS） and artesunate（ART） were prepared.The effect on inhibited NK killing of murine splenocytes was measured by MTT assay and that on inhibited CD3ε-ζ＋ expression was detected by flow cytometry of direct immunofluorescence labeling.The concentrations of six immunosuppressive molecules（TGF-β1,VEGF,IL-4,IL-6,IL-10 and PGE2） in supernatants were measured by quantitative ELISA.The results were compared with those of blank control.The multi-analysis of correlativity was used to ascertain the target molecules of each traditional Chinese medicine that could reverse the NK immunosuppressions of colorectal tumor cells.Results The down-regulating effect of AMB on inhibited NK killing was the strongest,and that of LHC on inhibited CD3ε-ζ＋ expression was the highest.The target molecule of As2O3,AMB,PUPS and ART on down-regulated inhibition of NK killing activity from Colon26 was TGF-β1,that of LHC was TGF-β1 and IL-10.The target molecule of AMB and MOX on down-regulated inhibition of CD3ε-ζ＋ expression was IL-10.Conclusion Reducing the immunosuppressions of NK cells caused by tumor cells through down-regulated immunosuppre-ssive molecules should be one of the mechanisms for traditional Chinese medicines to against tumor.It is reasonable to use different types of anti-tumor traditional Chinese medicines to inhibit NK cell immunosuppressions.

关 键 词：结直肠癌 免疫抑制 自然杀伤细胞 免疫抑制分子 

分 类 号：R285[医药卫生—中药学]