大鼠膝骨关节炎潜在生物标志物及代谢通路筛选  

Screening of potential biomarkers and metabolic pathways of knee osteoarthritis in rats

作　　者：王雨萌 王丽华[2] Wang Yumeng;Wang Lihua(Medical Record and Statistics Room of Ningbo Beilun People's Hospital,Ningbo 315800,China;Institute for Endemic Fluorosis Control,Center for Endemic Disease Control,Chinese Center for Disease Control and Prevention,Harbin Medical University,Harbin 150081,China)

机构地区：[1]浙江省宁波市北仑区人民医院病案统计室,315800 [2]哈尔滨医科大学中国疾病预防控制中心地方病控制中心地氟病防治研究所,150081

出　　处：《中华地方病学杂志》2021年第7期524-529,共6页Chinese Journal of Endemiology

基　　金：国家自然科学基金(81372937)。

摘　　要：目的筛选与大鼠膝骨关节炎相关的差异代谢物及其代谢通路,为深入研究骨关节炎生物标志物提供线索。方法60只雄性SPF级SD大鼠按体质量(300~350 g)采用随机数字表法分为模型组和对照组,每组30只。实验周期分别为4、8和12周,每个周期每组10只大鼠。模型组大鼠左侧膝关节采用改良Hulth法行造模手术,5 d后,驱赶大鼠使之活动,每天30 min。实验期间,两组大鼠均饲喂普通固体饲料、自由饮用自来水。实验期满,采集大鼠膝关节和血液样本。采用苏木素-伊红(HE)染色观察膝关节组织病理学变化,超高效液相色谱-四极杆飞行时间质谱仪(UPLC-Q-TOF-MS)检测血清中小分子代谢物,并通过多元统计分析和数据库比对,筛选与骨关节炎相关的差异代谢物及其相关代谢通路。结果模型组大鼠膝关节软骨变薄,表层粗糙、缺损或剥脱,软骨细胞变性、坏死、缺失,病变随实验周期延长而加重。筛选出11个与骨关节炎相关的血清差异代谢物,分别为硒代半胱氨酸、6-羟褪黑素、γ-谷氨酰半胱氨酸、花生四烯酸、二氢神经鞘氨醇、白三烯A4、白三烯B4、11,12-环氧-二十碳三烯酸(11,12-EpETrE)、溶血磷脂酰胆碱、神经酰胺和N-花生四烯酰甘氨酸。其中,实验4周时筛选出9个,与对照组比较,模型组5个高表达,4个低表达;实验8周时筛选出8个,与对照组比较,模型组2个高表达,6个低表达;实验12周时筛选出8个,与对照组比较,模型组5个高表达,3个低表达。筛选出2条与骨关节炎密切相关的代谢通路,分别为鞘脂代谢通路和花生四烯酸代谢通路,其中,二氢神经鞘氨醇和神经酰胺归属到鞘脂代谢通路,花生四烯酸、白三烯A4和白三烯B4归属到花生四烯酸代谢通路。结论骨关节炎疾病进程可以影响血清代谢物谱的构成和水平。11个血清差异代谢物涉及鞘脂代谢通路和花生四烯酸代谢通路,与骨关节炎的发生发展相�Objective To screen differential metabolites and metabolic pathways which are related to knee osteoarthritis in rats,and to provide clues for further study of biomarkers of osteoarthritis.Methods Sixty SPF male SD rats were divided into model and control groups according to their body weight(300-350 g)by random number table method,with 30 rats in each group.The experimental periods were 4,8 and 12 weeks,each period included 10 rats in each group.The left knee joints of rats in the model group were operated by the modified Hulth method.After 5 d,rats in the model group were driven to move for 30 min every day.All rats were fed ordinary solid fodder and drank tap water.At the end of the experimental period,the knee joints and blood samples of rats were collected.Hematoxylin-eosin(HE)staining was used to observe histopathological changes of knee joint,ultra-performance liquid chromatography quadrupole time-flight mass spectrometry(UPLC-Q-TOF-MS)was used to detect small molecule metabolites in serum,and multivariate statistical analysis and database comparison were used to screen the differential metabolites and related metabolic pathways associated with osteoarthritis.Results In the model group,the articular cartilage of knee was thinned,the surface was roughness,defect or peeling,chondrocytes were degeneration,necrosis and deletion,and the lesions were aggravated with prolonged experimental period.A total of 11 serum differential metabolites related to osteoarthritis were screened out,including selenocysteine,6-hydroxymelatonin,γ-glutamylcysteine,arachidonic acid,sphinganine,leukotriene A4,leukotriene B4,11,12-epoxy-eicostrienoic acid(11,12-EpETrE),lysopc,ceramide and N-arachidonoyl glycine.Among them,9 metabolites were screened out at 4 weeks,compared with the control group,5 metabolites were increased and 4 metabolites were decreased in the model group;8 metabolites were screened out at 8 weeks,compared with the control group,2 metabolites were increased and 6 metabolites were decreased in the model group;8 meta

关 键 词：骨关节炎 代谢组学 生物标志物 代谢通路 大鼠 

分 类 号：R684.3[医药卫生—骨科学] R-332[医药卫生—外科学]