世界中医药
文章摘要
引用本文:刘雷蕾,马淑然.基于肠道菌群介导的炎性反应研究黄芪多糖对结肠炎相关癌症的作用[J].世界中医药,2021,(02):.  
基于肠道菌群介导的炎性反应研究黄芪多糖对结肠炎相关癌症的作用
Study on the Mechanism of Astragalus Polysaccharide on Colitis-Related Cancer Based on the Inflammatory Response Mediated By Intestinal Flora
投稿时间:2019-09-16  
DOI:10.3969/j.issn.1673-7202.2021.02.008
中文关键词:  结直肠癌  肠道菌群  脂多糖  黄芪多糖  核因子κB
English Keywords:Colorectal carcinoma  Intestinal flora  Lipopolysaccharide  Astragalus polysaccharide  Nuclear factor κB
基金项目:中国博士后科学基金面上项目(2019M650596)
作者单位
刘雷蕾,马淑然 北京中医药大学,北京,100029 
摘要点击次数: 321
全文下载次数: 0
中文摘要:
      目的:探讨黄芪多糖(APS)对结肠炎相关癌(CAC)小鼠和结肠癌细胞中肠道细菌诱导的Toll样受体-4/核因子-κB(TLR4/NF-κB)信号通路的影响。方法:将40只C57BL/6J雄性小鼠随机分为对照组(n=10),模型组(n=15)和APS组(n=15)。模型组和APS组由氧化偶氮甲烷和葡聚糖硫酸钠制成CAC模型。对照组和模型组给予0.9%氯化钠注射液,APS组给予100 mg/kg APS灌胃14 d。动物研究中,通过HE染色方法检测小鼠中结肠直肠的病理状况;通过16S rDNA测序分析肠道菌群的特征;通过酶联免疫吸附试验测定脂多糖浓度;免疫组化法检测结肠组织中TLR4和NF-κBp65蛋白的表达。体外研究中,进一步验证了APS对人结肠癌细胞增殖和迁移的影响,运用MTT测定和划痕实验用于评估细胞活力测定;通过免疫荧光检测NF-κBp65的核转位;Western blot检测E-cadherin和N-cadherin蛋白的表达。结果:测序结果表明,模型组以革兰氏阴性菌拟杆菌属为主,APS组肠道菌群多为革兰氏阳性菌乳球菌属和双歧杆菌属;APS组LPS、TLR4和NF-κBp65蛋白的表达水平明显低于模型组;APS可以抑制LoVo细胞和HCT116细胞的增殖和迁移能力;APS(40 μg/mL)可抑制NF-κB的核转位,降低N-cadherin蛋白的表达,增加E-cadherin蛋白的表达。结论:APS通过抑制革兰氏阴性病原体,减少LPS的释放,抑制TLR4/NF-κB信号通路,阻止CAC的发生,且APS可有效抑制人结肠癌细胞的EMT过程,有效控制肿瘤的增殖和迁移。
English Summary:
      To explore the effects of Astragalus polysaccharide(APS)on intestinal bacteria induced Toll-like reeeptor-4 / nuclear factor-κB(TLR4/NF-κB)signaling pathway in colitis-associated cancer(CAC)mice and colon cancer cells.Methods:A total of 40 C57BL/6J male mice were randomLy divided into a control group(10 mice),a model group(15 mice),and an APS group(15 mice).The model group and APS group were made CAC model by azoxymethane and sodium dextran sulfate.The control group and model group were given 0.9% sodium chloride injection,and the APS group was given 100 mg/kg APS by gavage for 14 days.In this study,the Histopathological Examination method was used to detect the pathological conditions of colorectal in mice.Characteristics of intestinal flora were analyzed by 16S rDNA sequencing.Lipopolysaccharide concentrations were determined by enzyme 1inked immunosorbent assay.Expressions of TLR4 and NF-κB p65 proteins in colon tissue were detected by immunohistochemistry.In the in vitro study,we further validated the effects of APS on proliferation and migration of human colon cancer cells.The MTT assay and Scratch test were used to evaluate cell viability assays.The nuclear translocation of NF-κB p65 was detected by immunofluorescence.Western blot was used to detect the expression of E-cadherin and N-cadherin proteins.Results:Sequencing results showed that Gram-negative bacteria(Bacteroides)were commonly detected in the model group,and intestinal flora in APS group were mostly Gram-positive bacteria(Lactococcus and Bifidobacterium).Expression levels of LPS,TLR4,and NF-κB p65 proteins in APS group were significantly lower than those in the model group.APS can inhibit the proliferation and migration of LoVo cells and HCT116 cells; APS(40 μg/mL)can inhibit the nuclear translocation of NF-κB,reduce the expression of N-cadherin protein,and increase the expression of E-cadherin protein.Conclusion:APS can prevent CAC by inhibiting Gram-negative pathogens,reducing the release of LPS and inhibiting TLR4/NF-κB signaling pathway.And APS can effectively inhibit the EMT process of human colon cancer cells and effectively control tumor proliferation and migration.
查看全文  查看/发表评论  下载PDF阅读器