文献类型：期刊文献

中文题名：黄芪多糖对肺肿瘤转移前微环境中HIF-1和S100A8/A9蛋白表达及肺转移的影响

英文题名：Effect of Astragalus polysaccharides on HIF-1 and S100A8/A9 protein expression in the pre-metastatic niche of lung tumors and lung metastasis

作者：刘兆华[1,2,3,4];王彦君[5];申明[1,2,3,4];陈彦文[1];李杨[1,2,3,4];杨玲玲[1];梁乾坤[1];明海霞[1,2,3,4]

第一作者：刘兆华

机构：[1]甘肃中医药大学基础医学院,兰州730000;[2]甘肃中医药大学中西医结合基础学科,兰州730000;[3]甘肃中医药大学重大疾病分子医学与中医药防治研究重点实验室,兰州730000;[4]甘肃省中医药防治慢性疾病重点实验室,兰州730000;[5]甘肃中医药大学附属医院,兰州730000

第一机构：甘肃中医药大学基础医学院（敦煌医学研究所）

年份：2023

卷号：31

期号：7

起止页码：871

中文期刊名：中国实验动物学报

外文期刊名：Acta Laboratorium Animalis Scientia Sinica

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD_E2023_2024】；

基金：甘肃省“双一流”科研重点项目(GSSYLXM-05);甘肃省自然科学基金项目(20JR5RA178,20JR10RA320);甘肃省中医药研究中心开放课题(zyzx-2020-zx2);甘肃中医药大学科学研究与创新基金项目(2020KCZD-4,2021KCZD-1);甘肃中医药大学中西医结合基础学科科研培育项目(2022-2023-8,2023-2024-8);兰州市科技计划项目(2020-ZD-55,2021-1-96);甘肃省教育科技创新项目(2021CXZX-745)。

语种：中文

中文关键词：肺癌;黄芪多糖;芬戈莫德;肿瘤转移;转移前微环境

外文关键词：lung cancer;Astragalus polysaccharides;fingolimod;tumor metastasis;pre-metastatic niche

摘要：目的观察不同浓度黄芪多糖(Astragalus polysaccharides,APS)通过干预肺转移前微环境(Pre-metastatic niche,PMNs)对HIF-1和S100A8/A9蛋白的影响,初步阐明APS对肺PMNs中的作用靶点及对肿瘤肺转移影响的分子机制。方法SPF级C57BL/6J小鼠90只,除空白组外,其余75只用荧光素酶(LUC)标记的Lewis肺癌细胞1×10^(6)个通过小鼠尾静脉注射构建PMNs模型,并随机分成5组:模型组、低剂量组(灌服黄芪多糖50 mg/kg)、中剂量组(灌服黄芪多糖100 mg/kg)、高剂量组(灌服黄芪多糖200 mg/kg)、芬戈莫德(FTY720)组(腹腔注射FTY7201 mg/kg),每组15只。观察不同浓度黄芪多糖对PMNs中HIF-1和S100A8/A9蛋白的影响,同时用小动物活体成像及HE染色评估肺部转移情况。接种瘤细胞次日后低、中、高各剂量组按剂量灌胃给药,每天1次,共28 d。FTY720通过腹腔注射每2 d 1次间隔给药。在前14 d通过小动物活体成像动态观察肺肿瘤转移情况,并在第14天每组随机取出5只采用Western Blot及RT-qPCR技术检测多功能蛋白聚糖、纤维连接蛋白和赖氨酰氧化酶等在肿瘤转移前微环境中具有标志性的蛋白进一步验证模型成功。在第28天取肺组织采用苏木精-伊红(HE)染色观察小鼠肺组织病理学改变,并采用Western Blot及RT-qPCR技术检测28 d所取的肺组织中HIF-1和S100A8/A9蛋白和基因的表达,取材前12 h禁食不禁水。结果HE染色观察小鼠肺组织发现,与空白组相比模型组小鼠肺组织中出现大面积肺泡壁增厚和粒细胞浸润,并出现少量肿瘤转移灶;与模型组相比,高剂量组和FTY720组小鼠的肿瘤转移灶点明显减少,肺组织趋于正常。RT-qPCR、Western Blot检测结果显示:与空白组比较,模型组小鼠肺中HIF-1,S100A8,S100A9基因和蛋白表达显著升高(P<0.05);而与模型组相比,FTY720组和APS各组上述蛋白和基因表达均下降(P<0.05)。结论黄芪多糖可通过调控肺肿瘤转移前微环境中HIF-1、S100A8、S100A9的表达水平减少肿瘤细胞肺转移。
Objective To observe the effects of different concentrations of Astragalus polysaccharides(APS)on hypoxia-inducible factor(HIF)-1 and S100A8/A9 protein expression in the lung pre-metastatic niche(PMN),and to investigate the targets of APS in lung PMNs and its molecular mechanisms in relation to lung tumor metastasis.Methods Ninety specific-pathogen-free C57BL/6J mice were used,of which 15 were set as the blank control group.A PMN model was constructed in the remaining 75 mice by tail vein injection of 1×10^(6) luciferase-labeled Lewis lung cancer cells.These 75 mice were divided randomly into five groups(n=15 per group):model,low-dose(APS 50 mg/kg),medium-dose(APS 100 mg/kg),high-dose(APS 200 mg/kg),and fingolimod(FTY720,injected intraperitoneally 1 mg/kg)groups.APS was administered by gavage at the respective prescribed dose once a day on the day after inoculation of tumor cells and FTY720 was administered once every 2 days.The effects of different concentrations of APS on HIF-1 and S100A8/A9 proteins in the PMN was observed and lung metastasis was assessed by hematoxylin/eosin(HE)staining.On day 14,five mice from each group were randomly selected and lung tissues were removed and multifunctional proteoglycans,fibronectin,and lysyl oxidase were detected by Western Blot and Real-time qvantitative PCR(RT-qPCR),and histopathological changes in the lung were observed by HE staining.Protein and gene expression levels of HIF-1 and S100A8/A9 in lung tissues taken at 28 days were also detected by Western Blot and RT-qPCR.Mice were fasted but had access to water for 12 h before sampling.Results HE staining revealed alveolar wall thickening and granulocyte infiltration with a few tumor metastases in lung tissues in the model group.Tumor metastatic foci were significantly reduced and the lung tissues tended to be normal in the high-dose and FTY720 groups compared with the model group.HIF-1,S100A8,and S100A9 protein and gene expression levels were significantly higher in the lungs of mice in the model groups compared with the blank group(P<0.05),according to Western Blot and RT-qPCR.In contrast,the above protein and gene expression levels were decreased in the FTY720 group and all APS groups compared with the model group(P<0.05).Conclusions APS can reduce lung metastasis of tumor cells by regulating the expression levels of HIF-1,S100A8 and S100A9 in the PMN of lung tumors.