摘要
旨在开发一种组织渗透性好的35 kDa生物活性透明质酸片段(bioactive hyaluronic acid fragment,B-HA),并围绕着B-HA对白细胞移走与活性氧水平的影响展开研究,探究其临床抗炎机制,为B-HA在临床领域的应用提供试验依据。使用重组人透明质酸酶PH20切割高分子量透明质酸,制备低分子量35 kDa的透明质酸片段B-HA;使用伽马相机动态采集小鼠体内放射性元素标记的99m Tc-B-HA的组织分布;使用最接近人体临床研究的新鲜提取的人外周血中性粒细胞和单个核细胞(淋巴细胞为主),采用琼脂糖微滴法探究B-HA对人中性粒细胞和单个核细胞移走的影响。99m Tc-B-HA组织分布显示,B-HA静脉注射后血液半衰期仅为5 min,除绝大部分进入代谢器官肝脏,其余大部分分布在淋巴器官脾脏;300μg/mL的B-HA促进了新鲜提取的人外周血单个核细胞移走(P<0.05),300μg/mL的B-HA在有(P<0.001)和没有(0.001<P<0.05)内毒素LPS存在的情况下均明显抑制外周血中性粒细胞的移走,40μg/mL的B-HA抑制了人外周血中性粒细胞产生活性氧水平(P<0.01)。
This paper aims to develop a 35 kDa hyaluronic acid fragment B-HA with good tissue permeability and study the effects of B-HA on leukocyte migration and reactive oxygen species to explore its clinical anti-inflammatory mechanism and provide experimental basis for the application of B-HA in the clinical field.In this paper,a 35 kDa hyaluronic acid fragment B-HA was prepared by cutting high molecular weight hyaluronic acid with recombinant human hyaluronidase PH20.Gamma camera was used to dynamically capture the tissue distribution of 99 m Tc-B-HA labeled with radioactive elements in mice.The effects of B-HA on the migration of human neutrophils and mononuclear cells(mainly lymphocytes)from freshly extracted human peripheral blood were investigated using agrose-microdrip method,which was closest to human clinical studies.The tissue distribution of 99 m Tc-B-HA showed that the blood half-life of B-HA was only 5 min after intravenous injection,and most of the B-HA was distributed in the liver,the metabolic organ,and most of the rest were distributed in the spleen,the lymphatic organ;B-HA at 300μg/mL promoted the removal of freshly extracted human peripheral blood mononuclear cells(P<0.05),and B-HA(300μg/mL)significantly inhibited the migration of neutrophils both in the presence(P<0.001)and in the absence(0.001<P<0.05)of endotoxin LPS.B-HA(40μg/mL)inhibited the production of reactive oxygen species levels in human peripheral blood neutrophils(P<0.01).
作者
郭田田
王家麒
贾潇潇
惠觅宙
GUO Tiantian;WANG Jiaqi;JIA Xiaoxiao;HUI Mizhou(College of Animal Science and Technology,Qingdao Agricultural University,Qingdao 266109,China)
出处
《青岛农业大学学报（自然科学版）》
2023年第2期104-111,共8页
Journal of Qingdao Agricultural University(Natural Science)
基金
国家科技专项(2013ZX09J13110/04B-00)
青岛农业大学人才基金(665-1117001)。
关键词
透明质酸片段B-HA
组织分布
中性粒细胞
单个核细胞
移走
活性氧
hyaluronic acid fragments B-HA
tissue distribution
neutrophils
mononuclear cells
migration
reactive oxygen species
