益气养阴解毒方及其联合抗CTLA-4单抗对肺癌原位模型小鼠生存期及瘤体组织Foxp3、CTLA-4蛋白表达的影响  

Effect of Yiqi Yangyin Jiedu Formula(益气养阴解毒方) Combined with Anti-CTLA-4 Monoclonal Antibody on Survival Time and Foxp3 and CTLA-4 Protein Expression in Tumr Tissue of Orthotopic Lung Cancer Mice Model

作　　者：姜怡[1,2] 张朋 蔡雨晴[1,2] 阙祖俊 钱芳芳[4] 姚望 沈丽萍 田建辉[1,3] 刘苓霜[1] 刘嘉湘[1] JIANG Yi;ZHANG Peng;CAI Yuqing;QUE Zujun;QIAN Fangfang;YAO Wang;SHEN Liping;TIAN Jianhui;LIU Lingshuang;LIU Jiaxiang(Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai,200032;Graduate School,Shanghai University of Traditional Chinese Medicine;Shanhai Institute of Traditional Chinese Medicine and Cancer;Wuxi Huishan Traditional Chinese Medicine Hospital)

机构地区：[1]上海中医药大学附属龙华医院,上海市200032 [2]上海中医药大学研究生院 [3]上海市中医药研究院中医肿瘤研究所 [4]江苏省无锡市惠山区中医医院

出　　处：《中医杂志》2021年第10期908-913,共6页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(81703987);上海市卫生和计划生育委员会科研课题(201840158);上海市科学技术委员会科研项目(19401971200);刘嘉湘国医大师传承工作室建设项目(DSGZS-2017002)。

摘　　要：目的探讨益气养阴解毒方治疗肺癌的可能作用机制。方法将对数生长期的LLC-luc细胞于左肺注射建立C57BL/6小鼠肺癌原位模型。将造模成功的48只小鼠随机分为模型组,益气养阴解毒方低、中、高剂量组,单抗组,联合组,每组8只。模型组小鼠予生理盐水0.4ml灌胃及生理盐水0.3ml腹腔注射;益气养阴解毒方低、中、高剂量组分别给予益气养阴解毒方47、94、188g/(kg·d)灌胃及生理盐水0.3ml腹腔注射;单抗组予抗细胞毒T淋巴细胞相关抗原4(CTLA-4)单抗0.3ml腹腔注射及0.4ml生理盐水灌胃,共3次;联合组予益气养阴解毒方中剂量灌胃和抗CTLA-4单抗腹腔注射,用药剂量同上。各组均干预21天。活体成像动态监测小鼠的肿瘤生长并检测瘤体组织荧光强度,观察小鼠生存期。HE染色观察肿瘤细胞的形态,免疫组化法检测瘤体组织中叉头翼状螺旋转录因子P3(Foxp3)及CTLA-4蛋白表达。结果与模型组比较,益气养阴解毒方中、高剂量组及单抗组、联合组瘤体组织平均荧光强度均降低(P<0.05);联合组小鼠瘤体组织平均荧光强度低于益气养阴解毒方中、高剂量组和单抗组(P<0.05)。HE染色结果显示,模型组、益气养阴解毒方低剂量组小鼠瘤体组织见密集分布的低分化腺癌细胞,病理性核分裂象较多,益气养阴解毒方中、高剂量组和单抗组见病理性核分裂象减少,联合组见病理性核分裂象进一步减少。模型组,益气养阴解毒方低、中、高剂量组,单抗组和联合组小鼠的中位生存期分别为21、24、31、32、96、130天。与模型组比较,益气养阴解毒方中、高剂量组和单抗组、联合组均延长小鼠生存期,瘤体组织中Foxp3和CTLA-4蛋白表达均明显减少(P<0.05),且联合组较益气养阴解毒方中剂量组更能延长小鼠生存期(P<0.01),联合组小鼠瘤体组织Foxp3、CTLA-4蛋白表达较益气养阴解毒方高剂量组明显减少(P<0.05)。结论益气养阴Objective To explore the possible mechanism of Yiqi Yangyin Jiedu Formula(益气养阴解毒方,YYJF)in treatment of lung cancer.Methods LLC-luc cells in logarithmic period were injected into the left lung of C57 BL/6 mice to establish the model of orthotopic lung cancer.Forty-eight successfully modeled mice were randomized into model group,low,medium and high dose YYJF groups,monoclonal antibody group,and combination group with eight mice in each group.The mice in model group received 0.4 ml normal saline by gavage;the low,medium and high dose YYJF groups were given 47,94 and 188 g/(kg·d)YYJF by gavage respectively and intraperitoneal injection of 0.3 ml normal saline;the monoclonal antibody group received intraperitoneal injection of 0.3 ml anti-cytotoxic T lymphocyte-associated antigen 4(CTLA-4)monoclonal antibody and 0.4 ml normal saline by gavage,totally for three times;the combined group was administered with medium YYJF by gavage and intraperitoneal injection of anti CTLA-4 monoclonal antibody,with same dosage mentioned above;the intervention in all the groups lasted for 21 days.In vivo optical imaging system was used to monitor the tumor growth of mice and detect the fluorescence intensity in tumor tissue;survival time was assessed.HE staining method was applied to observe the morphology of tumor cells;immunohistochemical method was used to detect the protein expression of forkhead winged helix transcription factor P3(Foxp3)and CTLA-4.Results Compared to model group,the medium and high dose YYJF group,the monoclonal antibody group and the combined group had decreased mean fluorescence intensity(MFI)in tumor tissue(P<0.05),of which the combined group had lowest MFI(P<0.05).HE staining results showed that poorly differentiated adenocarcinoma cells were intensely distributed in the tumor tissues of model group and low dose YYJF group,and increased pathological mitotic figures were seen;the pathological mitotic figures decreased in medium and high dose YYJF groups and monoclonal antibody group,and decreased

关 键 词：肺癌 益气养阴解毒方 免疫逃逸 细胞毒T淋巴细胞相关抗原4 叉头翼状螺旋转录因子P3 

分 类 号：R285.5[医药卫生—中药学]