益气养阴方对Lewis肺癌移植模型小鼠免疫重塑相关因子表达的影响  

Effects of Yiqiyangyin Recipe on Immune Remodeling Related Cytokines in Lewis Lung Cancer Transplanted Mice

作　　者：胡丹丹[1] 杨国良[2] 陈素珍[1] 楼黎明[1] 张弘[1] HU Dandan;YANG Guoliang;CHEN Suzhen;LOU Liming;ZHANG Hong(Department of Respiratory Diseases,Third Affiliated Hospital,Zhejiang Chinese Medical University,Hangzhou,Zhejiang province,310005,China;Department of Oncology,Zhejiang Integrated Chinese and Western Medicine Hospital,Hangzhou,Zhejiang province,310003,China)

机构地区：[1]浙江中医药大学附属第三医院呼吸科,杭州310005 [2]浙江省中西医结合医院肿瘤科,杭州310003

出　　处：《浙江中西医结合杂志》2020年第2期96-100,I0002,共6页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

基　　金：浙江省自然科学基金青年项目(No.LQ18H270002);“十三五”浙江省中医药(中西医结合)重点学科建设计划(No.2017-XK-A19);浙江省中医药科技计划项目(No.2016ZA108);浙江省中医药科技计划项目(No.2015ZB084)。

摘　　要：目的观察益气养阴方对Lewis肺癌移植模型小鼠免疫重塑相关因子表达的影响。方法30只近交系C57BL/6纯系小鼠行Lewis肺癌细胞移植造模后随机分为三组:模型组、中药组及西药组,每组10只。模型组仅造模;中药组造模后予灌胃益气养阴方悬液(含生药2g/mL),1天1次,每次0.4mL;西药组将Nivolumab按照0.273mg/10g溶于0.2mL生理盐水中腹腔注射,每2周1次;各组均常规喂食。造模后第2天开始给药,给药3周。第7天开始观察记录各组小鼠瘤体大小(每3天测1次瘤体大小)。观察期满后各组小鼠断颈处死,留取各组小鼠肿瘤组织,ELISA法测定肿瘤组织匀浆白细胞介素-10(IL-10)、转化生长因子-β(TGF-β)、转录因子p3(Foxp3)含量,流式细胞仪检测小鼠外周血CD4+CD25+Treg细胞比例。结果中药组及西药组肿瘤体积增长均减慢,两组抑瘤率比较(23.07%比37.55%),差异无统计学意义(P>0.05);与模型组比较,中药组及西药组肿瘤组织IL-10含量(1915.79±114.13)pg/mL、(1712.71±157.25)pg/mL比(2257.78±63.00)pg/mL;TGF-β含量(99.59±18.64)pg/mL、(66.83±9.75)pg/mL比(142.33±8.16)pg/mL;Foxp3表达(0.83±0.11)、(0.35±0.09)比(0.98±0.08),均明显下降(P均<0.05);与中药组比较,西药组肿瘤组织IL-10含量(1712.71±157.25)pg/mL比(1915.79±114.13)pg/mL,TGF-β含量(66.83±9.75)pg/mL比(99.59±18.64)pg/mL,Foxp3表达(0.35±0.09)比(0.83±0.11),均明显降低(P均<0.05);与模型组比较,中药组及西药组小鼠外周血CD4+CD25+Treg细胞比例(1.07±0.12)%、(0.50±0.26)%比(1.17±0.38)%,明显降低(P<0.05),西药组小鼠外周血CD4+CD25+Treg细胞比例(0.50±0.26)%较中药组(1.07±0.12)%降低更明显(P<0.05)。结论益气养阴方可抑制Lewis肺癌移植模型小鼠肿瘤生长,其机制可能与降低肿瘤组织Foxp3表达,减少CD4+CD25+Treg细胞分化,降低IL-10、TGF-β表达,改善细胞免疫抑制状态相关。Objective To observe the effects of Yiqiyangyin(benefiting qi and nourishing yin) recipe on immune remodeling related cytokines in Lewis lung cancer transplanted mice. Methods Thirty inbred C57 BL/6 pure mice were transplanted with Lewis lung cancer cells to establish the disease models, then the mice were randomly divided into three groups: model group, traditional Chinese medicine group and western medicine group, with 10 animals in each group. The model group was treated without anything. In the traditional Chinese medicine group, suspen-sion of Yiqiyangyin recipe including 2 g/mL crude drug was administrated once a day, 0.4 mL each time. In the western medicine group, Nivolumab dissolved in 0.2 mL normal saline was injected intraperitoneally at a dose of0.273 mg/10 g, once every two weeks. All nice were fed regularly. The drug was administered for 3 weeks on the second day after modeling. The tumor size of mice in each group was measured and recorded from the 7 th day,and once every 3 days. At the end of the observation period, mice in each group were sacrificed and tumor tissues were collected. The levels of interleukin-10(IL-10), transforming growth factor-β(TGF-β) and transcription factor p3(Foxp3) in tumor tissue homogenate were determined by ELISA. The ratio of CD4+CD25+Treg cells in peripheral blood was detected by flow cytometry. Results The growth of tumor volume in both the traditional Chinese medicine group and the western medicine group slowed down, and there was no significant difference in tumor inhibition rate between the two groups(23.07% vs 37.55%). Compared to the model group, expression of IL-10,TGF-β and Foxp3 decreased significantly, i.e., IL-10: [(1915.79±114.13) and(1712.71±157.25) vs(2257.78±63.00)pg/mL, P<0.05], TGF-β: [(99.59±18.64) and(66.83±9.75) vs(142.33±8.16)pg/mL, P<0.05], Foxp3: [(0.83±0.11) and(0.35 ±0.09) vs(0.98 ±0.08), P <0.05]. But the expression of IL-10 [(1712.71 ±157.25) vs(1915.79 ±114.13), P<0.05], TGF-β[(66.83±9.75) vs(99.59±18.64), P<0.05]、Foxp3[

关 键 词：小鼠 肺癌 免疫重塑 IL-10 TGF-β CD4+CD25+TREG细胞 FOXP3 益气养阴 

分 类 号：R285.5[医药卫生—中药学] R-332[医药卫生—中医学]