400038 重庆,陆军军医大学(第三军医大学)药学与检验医学系
Received:19 July 2023,
Revised:2023-08-30,
Published:15 March 2024
移动端阅览
Yaoyao LIU, Juanjuan LIU, Yiyu QIN, et al. Affinity test of 18 traditional Chinese medicine polysaccharides with Siglec-9 and their anti-inflammatory effects[J]. Journal of Army Medical University, 2024, 46(5): 410-418.
Yaoyao LIU, Juanjuan LIU, Yiyu QIN, et al. Affinity test of 18 traditional Chinese medicine polysaccharides with Siglec-9 and their anti-inflammatory effects[J]. Journal of Army Medical University, 2024, 46(5): 410-418. DOI: 10.16016/j.2097-0927.202307103.
目的
2
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
2
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
2
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
2
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
2
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
2
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
2
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
2
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.
HOU C Y, CHEN L L, YANG L Z, et al . An insight into anti-inflammatory effects of natural polysaccharides[J ] . Int J Biol Macromol, 2020, 153: 248-255. DOI: 10.1016/j.ijbiomac.2020.02.315.
舒任庚, 蒋跃平, 蔡永红. 植物多糖的提取分离方法探讨[J]. 中国药房, 2011, 22(11): 1052-1055.
SHU R G, JIANG Y P, CAI Y H. Exploration of extraction and isolation method of plant polysaccharides[J]. China Pharm, 2011, 22(11): 1052-1055.
YU Y, SHEN M Y, SONG Q Q, et al . Biological activities and pharmaceutical applications of polysaccharide from natural resources: a review[J ] . Carbohydr Polym, 2018, 183: 91-101. DOI: 10.1016/j.carbpol.2017.12.009.
JIANG P, JI X, XIA J, et al . Structure and potential anti-fatigue mechanism of polysaccharides from Bupleurum chinense DC[J ] . Carbohydr Polym, 2023, 306: 120608. DOI: 10.1016/j.carbpol.2023.120608..
GAO W D, FENG Z J, ZHANG S L, et al . Anti-inflammatory and antioxidant effect of Eucommia ulmoides polysaccharide in hepatic ischemia-reperfusion injury by regulating ROS and the TLR-4-NF- κ B pathway[J ] . Biomed Res Int, 2020, 2020: 1860637. DOI: 10.1155/2020/1860637..
GAO Q H, FU X Y, ZHANG R, et al . Neuroprotective effects of plant polysaccharides: a review of the mechanisms[J ] . Int J Biol Macromol, 2018, 106: 749-754. DOI: 10.1016/j.ijbiomac.2017.08.075.
王青, 苏聪平, 张惠敏, 等. 从炎性反应角度探讨清热解毒药的作用机制[J]. 中国中药杂志, 2018, 43(18): 3787-3794. DOI: 10.19540/j.cnki.cjcmm.20180611.012.
WANG Q, SU C P, ZHANG H M, et al . Anti-inflammatory mechanism of heat-clearing and detoxifying Chinese herbs[J ] . China J Chin Mater Med, 2018, 43(18): 3787-3794. DOI: 10.19540/j.cnki.cjcmm.20180611.012.
蒋毅萍. 清热药物的清热、解毒、抗炎作用的探讨[D]. 广州: 南方医科大学, 2012.
JIANG Y P. Discussion on heat-clearing, detoxification and anti-inflammatory effects of heat-clearing drugs[D]. Guangzhou: Southern Medical University, 2012.
ZENG P J, LI J, CHEN Y L, et al . The structures and biological f unctions of polysaccharides from traditional Chinese herbs[J ] . Prog Mol Biol Transl Sci, 2019, 163: 423-444. DOI: 10.1016/bs.pmbts.2019.03.003.
SMITH B A H, BERTOZZI C R. The clinical impact of glycobiology: targeting selectins, Siglecs and mammalian glycans[J]. Nat Rev Drug Discov, 2021, 20(3): 217-243. DOI: 10.1038/s41573-020-00093-1.
MURUGESAN G, WEIGLE B, CROCKER P R. Siglec and anti-siglec therapies[J]. Curr Opin Chem Biol, 2021, 62: 34-42. DOI: 10.1016/j.cbpa.2021.01.001.
LIZCANO A, SECUNDINO I, DÖHRMANN S, et al . Erythrocyte sialoglycoproteins engage Siglec-9 on neutrophils to suppress activation[J ] . Blood, 2017, 129(23): 3100-3110. DOI: 10.1182/blood-2016-11-751636.
HSU Y W, HSU F F, CHIANG M T, et al . Siglec-E retards atherosclerosis by inhibiting CD36-mediated foam cell formation[J ] . J Biomed Sci, 2021, 28(1): 5. DOI: 10.1186/s12929-020-00698-z.
SIDDIQUI S, SCHWARZ F, SPRINGER S, et al . Studies on the detection, expression, glycosylation, dimerization, and ligand binding properties of mouse siglec-E[J ] . J Biol Chem, 2017, 292(3): 1029-1037. DOI: 10.1074/ jbc.M116.738351.
THIESLER H, BEIMDIEK J, HILDEBRANDT H. Polysialic acid and Siglec-E orchestrate negative feedback regulation of microglia activation[J]. Cell Mol Life Sci, 2021, 78(4): 1637-1653. DOI: 10.1007/s00018-020-03601-z.
DELAVERIS C S, WILK A J, RILEY N M, et al . Synthetic siglec-9 agonists inhibit neutrophil activation associated with COVID-19[J ] . ACS Cent Sci, 2021, 7(4): 650-657. DOI: 10.1021/acscentsci.0c01669.
CAO P, WU S L, WU T T, et al . The important role of polysaccharides from a traditional Chinese medicine-Lung Cleansing and Detoxifying Decoction against the COVID-19 pandemic[J ] . Carbohydr Polym, 2020, 240: 116346. DOI: 10.1016/j.carbpol.2020.116346.
SECUNDINO I, LIZCANO A, ROUPÉ K M, et al . Host and pathogen hyaluronan signal through human siglec-9 to suppress neutrophil activation[J ] . J Mol Med, 2016, 94(2): 219-233. DOI: 10.1007/s00109-015-1341-8.
曾小燕, 李鸿霞, 梁纪强, 等. 中药多糖体外抗氧化作用研究进展[J]. 亚太传统医药, 2020, 16(7): 181-188. DOI: 10.11954/ytctyy.202007060.
ZENG X Y, LI H X, LIANG J Q, et al . Research progress on polysaccharide of traditional Chinese medicine anti-oxidant in vitro experiments[J ] . Asia Pac Tradit Med, 2020, 16(7): 181-188. DOI: 10.11954/ytctyy.202007060..
摄渊婷, 曾东风, 刘红梅, 等. 缺氧下调血小板表面Siglec-9聚糖配体导致血小板功能异常活化[J]. 第三军医大学学报, 2022, 44(6): 541-548. DOI: 10.16016/j.2097-0927.202111025.
SHE Y T, ZENG D F, LIU H M, et al. Hypoxia-downregulated Siglec-9 glycan ligands induce abnormal activation of platelet[J]. J Third Mil Med Univ, 2022, 44(6): 541-548. DOI: 10.16016/j.2097-0927.202111025.
LIU H M, ZHENG Y, ZHANG Y X, et al . Immunosuppressive Siglec-E ligands on mouse aorta are up-regulated by LPS via NF-κB pathway[J ] . Biomed Pharmacother, 2020, 122: 109760. DOI: 10.1016/j.biopha.2019.109760..
MIRALDA I, SAMANAS N B, SEO A J, et al . Siglec-9 is an inhibitory receptor on human mast cells in vitro [J ] . J Allergy Clin Immunol, 2023, 152(3): 711-724. e14. DOI: 10.1016/j.jaci.2023.04.007..
0
Views
0
下载量
0
CSCD
Publicity Resources
Related Articles
Related Author
Related Institution
京公网安备11010802024621
