白芨多糖对LPS诱导的巨噬细胞炎症模型中促炎因子的影响及机制研究  

Bletilla polysaccharides in the cell model of TLR4/nf-kappa B signaling pathways inhibitory effect and mechanism research

作　　者：张清[1] 蔡露[1] 陈波[1] 罗磊[1] 汪建军 ZHANG Qing;CAI Lu;CHEN Bo;LUO Lei;WANG Jian-jun(The Second People's Hospital in Yichang of Hubei,443000 Yichang,Hubei,China;Yunmeng County People's Hospital,Xiaogan City,Hubei Province,432500 Yunmeng,Hubei,China)

机构地区：[1]湖北省宜昌市三峡大学第二人民医院,湖北宜昌443000 [2]湖北省孝感市云梦县人民医院,湖北云梦432500

出　　处：《临床消化病杂志》2023年第6期447-452,共6页Chinese Journal of Clinical Gastroenterology

基　　金：宜昌市科技计划项目(No:A19-301-37)。

摘　　要：[目的]通过不同浓度白芨多糖干预内毒素激活后的小鼠RAW264.7巨噬细胞株,明确白芨多糖在细胞模型中对巨噬细胞TLR4信号通路中相关分子的干预作用。[方法]采用CCK-8法测定不同浓度白芨多糖溶液对Raw264.7巨噬细胞株的最大无毒浓度;采用ELISA法检测正常对照组,内毒素组,高、中、低浓度白芨多糖组,美沙拉嗪组促炎细胞因子TNF-α、IL-6、IL-1β和IL-17的表达水平;采用Real-time PCR法和Western blot法检测各组TLR4、MyD88、IRAK1、IKK-β、IRF3、IκB-α、NF-κB的mRNA和蛋白的表达情况。采用免疫荧光法检测各组TLR4和NF-κB的表达情况。[结果]内毒素组的各促炎细胞因子相较于正常对照组的促炎细胞因子明显升高,具有显著差异(P<0.05或0.01)。而白芨多糖作用于内毒素组巨噬细胞后,其促炎细胞因子、TLR4的mRNA表达量和蛋白含量及TLR4介导的下游MyD88,IKK-β,IκB-α和NF-κB分子mRNA表达量和蛋白含量均显著下降(P<0.05或0.01)。[结论]白芨多糖可通过拮抗内毒素诱导的巨噬细胞TLR4/NF-κB通路的活化,而影响促炎细胞因子的表达。[Objective]To determine the effect of Bletilla striata polysaccharide on TLR4 signaling path-way of macrophages in mouse RAW264.7 macrophages activated by endotoxin with different concentra-tions.[Methods]The maximum non-toxic concentration of Bletilla striata polysaccharide on Raw264.7 mac-rophage cell line was determined by CCK-8 method.The expression levels of pro-inflammatory cytokines TNF-α,IL-6,IL-1βand IL-17 in normal control group,endotoxin group,high,medium and low concentra-tion Bletilla striata polysaccharide group and mesalazine group were detected by ELISA.Real-time PCR and Western blot were used to detect the mRNA and protein expressions of TLR4,MyD88,IRAK1,IKK-β,IRF3,IsB-α,NF-kB in each group.The expression of TLR4 and NF-B in each group was detected by im-munofluorescence method.[Results]The levels of proinflammatory cytokines in endotoxin group were sig-nificantly higher than those in normal control group(P<0.05 or 0.01).The mRNA expression and protein content of proinflammatory cytokines,TLR4 and TLR4-mediated downstream MyD88,IKK-β,IB-αand NF-kB molecules were significantly decreased after treatment with Bletilla striata polysaccharide(P<0.05 or 0.01).[Conclusion]Bletilla striata polysacharide can affect the expression of proinflammatory cytokines by antagonizing the activation of TLR4/NF-sB pathway in macrophages induced by endotoxin.

关 键 词：结肠炎 溃疡性 白芨多糖 内毒素 TLR4 促炎细胞因子 

分 类 号：R574.62[医药卫生—消化系统]