毛钩藤碱调控Shh信号通路对结直肠癌荷瘤鼠的抑瘤作用及CD4^(+)、CD8^(+)T细胞的影响  

Anti-tumor effect of hirsutine regulated Shh signaling pathway on colorectal cancer-bearing mice and effect of CD4^(+)and CD8^(+)T cells

作　　者：王晨宇[1] 李兴旺 张军杰[1] 姚坤厚[1] 华龙[1] 胡军红[1] WANG Chenyu;LI Xingwang;ZHANG Junjie;YAO Kunhou;HUA Long;HU Junhong(Department of General Surgery,Huaihe Hospital,Henan University,Kaifeng 475000,China)

机构地区：[1]河南大学淮河医院普通外科一病区,开封475000

出　　处：《中国免疫学杂志》2022年第20期2473-2478,共6页Chinese Journal of Immunology

基　　金：河南省医学科技攻关计划项目(2018020331,212102310696,222102310729)。

摘　　要：目的:探讨毛钩藤碱对结直肠癌荷瘤鼠的抑瘤作用,并分析其对Shh信号通路关键蛋白及CD4^(+)、CD8^(+)T细胞的影响。方法:采用结肠癌SW480细胞接种于裸鼠右腋部皮下建立结直肠癌荷瘤鼠模型,将40只建模成功的裸鼠分为荷瘤鼠模型组、阳性药环巴胺(Shh信号通路抑制剂)对照组和毛钩藤碱低、中、高剂量组,每组8只。阳性药给予环巴胺(20μmol/L,0.2 ml),毛钩藤碱干预组分别腹腔注射5、10、20 mg/kg毛钩藤碱,1次/d,连续21 d,留取外周血后处死。观察各组荷瘤小鼠的自主活动次数、体质量及肿瘤体积,计算体质量增长率、相对肿瘤体积增长率和抑瘤率;免疫组化法(IHC)检测荷瘤鼠移植瘤中Glil、Gli2、Shh、Smo和Ptch1表达量;流式细胞术检测荷瘤鼠移植瘤和外周血中CD4^(+)及CD8^(+)T细胞水平。结果:与模型组比较,环巴胺及毛钩藤碱的干预显著改善了裸鼠的自主活动次数,环巴胺组及毛钩藤碱中、高剂量组的瘤质量、相对肿瘤体积增长率及Shh信号通路关键蛋白Glil、Gli2及Shh、Smo和Ptch1表达量均显著降低(P<0.05或P<0.01),移植瘤和外周血中CD4^(+)及CD8^(+)T细胞水平均显著提高(P<0.05或P<0.01),但毛钩藤碱低剂量组上述指标与模型组比较差异无统计学意义(P>0.05)。与阳性药环巴胺组比较,毛钩藤碱各剂量干预组肿瘤质量、抑瘤率、相对肿瘤体积增长率差异均有统计学意义(P<0.05或P<0.01)。随着毛钩藤碱剂量的增加,Shh信号通路关键蛋白(Glil、Gli2及Shh、Smo和Ptch1)表达量显著降低(P<0.05或P<0.01),而移植瘤和外周血中CD4^(+)及CD8^(+)T细胞水平均显著增加(P<0.05或P<0.01)。结论:毛钩藤碱可显著改善结肠癌荷瘤鼠的自主活动状态,发挥较好的抑瘤作用,可能与其抑制Shh信号通路,提高CD4^(+)及CD8^(+)T细胞水平有关。Objective:To investigate the anti-tumor effect of hirsutine on colorectal cancer-bearing mice,and to analyze its influence on key proteins of Shh signaling pathway and CD4^(+)and CD8^(+)T cells.Methods:Colon cancer SW480 cells were inoculated subcutaneously into the right axillary of nude mice to establish a colorectal cancer tumor-bearing mouse model.Forty nude mice that were successfully modeled were divided into tumor-bearing mouse model group and the positive drug cyclopamine(Shh signaling pathway inhibitor)control group and low,medium and high dose of hirsutine groups,with 8 rats in each group.Cyclopamine(20μmol/L,0.2 ml)was given to the positive drug,and the hirsutine intervention groups were given 5,10,20 mg/kg hirsutine,all the nude mouse were given once a day for 21 consecutive days,peripheral blood was collected and then sacrificed.The number of autonomous activities,body weight and tumor volume of tumor-bearing mice in each group were observed,and the growth rate of body weight,relative tumor volume growth rate and tumor inhibition rate were calculated.Immunohistochemistry(IHC)was used to detect the expressions of Glil,Gli2,Shh,Smo and Ptch1 in the transplanted tumor of tumor-bearing mice.Flow cytometry was used to detect the levels of CD4^(+)and CD8^(+)T cells in the transplanted tumor and peripheral blood of tumor-bearing mice.Results:Compared with the model group,the intervention of cyclopamine and hirsutine significantly improved the number of autonomous activities of nude mice,the tumor mass and relative tumor volume growth rate of the cyclopamine group and the middle and high-dose hirsutine groups and Shh signaling pathway key proteins Glil,Gli2 and Shh,Smo and Ptch1 expression were significantly reduced(P<0.05 or P<0.01),the level of CD4^(+)and CD8^(+)T cells in the transplanted tumor and peripheral blood increased significantly(P<0.05 or P<0.01),but the above indicators of the low-dose hirsutine group were not significantly different from the model group(P>0.05).Compared with the positive drug

关 键 词：毛钩藤碱 结直肠癌 荷瘤鼠 SHH信号通路 CD4^(+) CD8^(+) 

分 类 号：R285.5[医药卫生—中药学]