灵芝提取物在肿瘤免疫治疗中的调节作用  

Study on the regulatory efficacy of ganoderma lucidum extract to tumor immunotherapy

作　　者：朱林波 傅庆国[2] 刘卫怀 孙明非 李鹏飞 张建辉 张远炎 李吉 李晔 

机构地区：[1]浙江省宁波市北仑区人民医院普外科,315800 [2]浙江省宁波市第二医院肿瘤外科,315010 [3]福建仙芝楼生物科技有限公司,350002

出　　处：《中华全科医学》2014年第3期341-343,共3页Chinese Journal of General Practice

基　　金：浙江省中医药科技计划项目(2006B117)

摘　　要：目的将灵芝提取物与肿瘤免疫制剂联合运用,探讨灵芝提取物在肿瘤免疫治疗中的调节作用。方法纯化Hepa 1-6细胞的肿瘤热休克蛋白70多肽复合物(HSP70-PCs),SDS-PAGE及Western-blot对产物进行鉴定。将Hepa 1-6细胞浓度调整为4×107/ml,按0.1 ml/只接种于C57BL/6j小鼠背部皮下建立实验模型。随机将模型分为PBS对照组、灵芝提取物组(L组)、HSP70-PCs组(H组)、CpG-ODN组(C组)、灵芝提取物联合HSP70-PCs组(L+H组)和灵芝提取物联合CpG-ODN组(L+C组),每组10只。L组、L+C组、L+H组于建模的第7天开始予灵芝提取物按4 g/kg体重灌胃,1次/d,直至实验组小鼠死亡或实验结束;C组和L+C组予瘤旁皮下注射CpG-ODN 100μg/只,每周2次,共8次;H组和L+H组予瘤体对侧背部皮下注射HSP70-PCs 10μg/只,每周1次,共4次。对照组予皮下注射灭菌PBS液0.1 ml/只,每周1次,共4次。定时测量瘤体大小,记录肿瘤生长情况、有无转移灶及生存时间。ELISA法检测实验鼠血清CXCL-9、CXCL-10、IFN-γ水平。运用SPSS 16.0软件进行数据处理。结果灵芝提取物联合CpG-ODN或HSP70-PCs可以诱导抑瘤作用,肿瘤体积、质量、肿瘤转移率及生存期限与单独使用HSP70-PCs或CpG-ODN比较差异有统计学意义(P<0.05或P<0.01);灵芝提取物联合CpGODN或HSP70-PCs可以提高荷瘤小鼠血清中CXCL-9、CXCL-10和IFN-γ水平,与二者单独使用组比较差异有统计学意义(P<0.05或P<0.01)。结论灵芝提取物可通过诱导Th1型细胞因子表达来发挥免疫调节作用,对肿瘤免疫治疗有增强作用。Objective To investigate the regulatory efficacy of ganoderma lucidum extract （GLE） to tumor immunotherapy by combining GLE and HSP70-PCs or CpG-ODN. Methods HSP70-PCs was purified from the Hepa 1-6,then was identified by SDS-PAGE and Western-blot. The concentration of Hepa 1 -6 cells were adjusted to 4×10^7/ml, and 0.1 ml of the cell suspension was injected subcutaneously in the back of C57BL/6j mice. Then the model were randomly divided into 6 groups ： the PBS group, the GLE group （ L group ）, the HSP70-PCs group （ H group）, the CpG-ODN group （ C group）, the GLE ＋ HSP70-PCs group （L ＋ H group）, and the GLE ＋ CpG-ODN group （L ＋ C group）, with 10 mice in each group; GLE at the dose of 4 g/kg was given to L group, L ＋ H group and L ＋ C group by gastrogavage once a day until the experiment ended or the mice dead. CpG-ODN at the dose of 100 ug via subcutaneously injection to the C group and the L ＋ C group twice a week,and total 8 times;HSP70-PCs at the dose of 10 ug via subcutaneously injection to the H group and the L ＋ H group once a week,with a total of 4 times. Equal volume of PBS was given to the PBS group. The tumor volume, weight, metastasia rate, and average life span of mice of each group were recorded. Cytokines （ CXCL-9, CXCL-10 and IFN-γ） levels were assessed by ELISA. The data was processed by SPSS 16.0 software. Results Combination of GLE and CpG-ODN or HSP70-PCs was more effective than either of the two therapeutic agents alone. The tumor volume, weight, metastasia rate, average life span of mice of co-use group were significantly different between the group of HSP70- PCs and group of CpG-ODN. The levels of serum three cytokines in co-use groups were notably promoted than those of two agents administrated alone. Conclusion GLE appeared regulatory efficacy and promoted immunotherapy via enhencing the serum Thl cytokines levels.

关 键 词：灵芝提取物 免疫治疗 肿瘤 

分 类 号：R979.5[医药卫生—药品] R730.51[医药卫生—药学]