摘要
目的比较18种中药多糖与唾液酸特异性Ig样凝集素-9(sialic-acid-binding immunoglobulin-like lectin,Siglec-9)的亲和力以及抗炎能力,探讨中药多糖发挥抗炎作用的新机制。方法选择18种不同药性的中药用水提醇沉法提取多糖,苯酚硫酸法检测多糖含量,高效液相色谱法检测多糖的单糖组成,红外光谱法检测中药多糖结构。通过竞争性ELISA法检测多糖与Siglec-9的亲和力,并研究中药多糖对LPS诱导巨噬细胞炎症反应以及R848诱导中性粒细胞NETosis的影响,利用髓系细胞特异性敲除Siglec-E(Lyz2-cre:Siglec-E flox/flox)小鼠来源的原代巨噬细胞和腹腔中性粒细胞进一步明确多糖结合Siglec-9产生抗炎作用的机制。结果中药多糖的糖含量均在55%以上,主要由甘露糖、葡萄糖、半乳糖、阿拉伯糖、葡萄糖醛酸等组成,但单糖构成比各不相同。红外光谱检测4种代表性中药多糖均有典型的多糖吸收峰,柴胡和蒲公英多糖具有葡萄糖醛酸结构。枸杞和黄芪多糖与Siglec-9的亲和力,对LPS诱导的巨噬细胞炎症反应和R848诱导的中性粒细胞NETosis反应无显著抑制作用。柴胡和蒲公英多糖与Siglec-9亲和力较强(P<0.01),能够显著抑制上述炎症反应,但其抗炎作用在髓系细胞特异性敲除Siglec-E基因小鼠来源的巨噬细胞和中性粒细胞中消失。结论中药多糖与Siglec-9的亲和力有潜力成为研究中药抗炎作用的新的现代药理学机制。
Objective To compare the affinity with sialic acid-binding immunoglobulin-like lectin-9(Siglec-9)and anti-inflammatory effects of 18 traditional Chinese medicine polysaccharides in order to explore a new mechanism for the anti-inflammatory effects of these polysaccharides.Methods Aqueous alcohol precipitation was used to extract polysaccharides from 18 traditional Chinese medicines with different medicinal properties.Phenol sulfuric acid method was employed to detect the content of the polysaccharides,high performance liquid chromatography(HPLC)was utilized to measure the monosaccharide composition of the polysaccharides,and infrared spectroscopy was applied to detect the structure of the polysaccharides of Chinese medicines.The affinity of polysaccharide to Siglec-9 was detected by competitive ELISA,and the effects of the polysaccharides on LPS-induced inflammatory responses in macrophages and R848-induced NETosis in neutrophils were investigated.Primary cultured macrophages and neutrophils derived from myeloid-specific Siglec-E knockout mice(Lyz2-cre:Siglec-E flox/flox)were used to further clarify the mechanism of the anti-inflammation of the polysaccharides.Results All these 18 traditional Chinese medicines had the content of polysaccharide over 55%,and mainly composed of mannose,glucose,galactose,arabinose and glucuronic acid,with different composition ratio of monosaccharide separately.Infrared spectroscopy revealed that there were typical polysaccharide absorption peaks in all 4 representative polysaccharides,and the polysaccharides from Bupleurum chinense and Dandelion had glucuronic acid structures.The affinities with Siglec-9 in the polysaccharides from Lycium barbarum and Astragalus showed no inhibitory effect on LPS-induced inflammatory responses in macrophages and R848-induced NETosis in neutrophils.While,Bupleurum chinense and dandelion polysaccharides presented strong affinities with Siglec-9(P<0.01),and exerted obvious inhibitory effect on above inflammatory reactions,but the anti-inflammatory effects were lost disappeared in the macrophages and neutrophils derived from Siglec-E conditional knockout mice.Conclusion The affinity between Chinese herbal polysaccharide and Siglec-9 has a possibility to be a potential novel pharmacological mechanism of the anti-inflammatory effects of traditional Chinese medicine.
作者
刘瑶瑶
刘娟娟
秦怡玉
李瑾
刘红梅
贾乙
LIU Yaoyao;LIU Juanjuan;QIN Yiyu;LI Jin;LIU Hongmei;JIA Yi(Faculty of Pharmacy and Laboratory Medicine,Army Medical University(Third Military Medical University),Chongqing,400038,China)
出处
《陆军军医大学学报》
CAS
CSCD
北大核心
2024年第5期410-418,共9页
Journal of Army Medical University
基金
国家自然科学基金面上项目(81973319)
重庆市自然科学基金面上项目(cstc2019jcyj-msxmX0372)。
关键词
中药多糖
Siglec-9
抗炎
NETosis
traditional Chinese medicine polysaccharide
Siglec-9
anti-inflammation
NETosis
