T-2毒素致关节软骨损伤大鼠尿液代谢组学分析  

Urinary metabolic profiling of rats with articular cartilage damage induced by T-2 toxin

作　　者：朱雷[1] 赵志军[2] 刘施雨 王为一[1] 胡健[1] 李强[1] 王丽华[1] 

机构地区：[1]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,哈尔滨150081 [2]青海省地方病预防控制所地方病预防控制科,西宁811602

出　　处：《中华地方病学杂志》2016年第5期350-353,共4页Chinese Journal of Endemiology

基　　金：国家自然科学基金（81372937）

摘　　要：目的寻找T-2毒素致关节软骨损伤大鼠尿液中与关节软骨损伤相关的代谢物。方法将30只4～5周龄的雄性Wistar大鼠．按体质量采用随机数字表法，将大鼠分为两组：对照组和模型组，每组15只。对照组大鼠饲喂普通饲料，模型组大鼠饲喂经T-2毒素染毒（300斗g／kg）饲料，实验期间，两组大鼠自由进食和饮水。3个月后处死大鼠，取膝关节软骨进行组织病理学观察，利用超高效液相色谱．四级杆．飞行时间质谱（UPLC／QTOF．MS）技术检测尿液代谢物谱。通过多元统计分析以及数据库比对，寻找与关节软骨损伤相关的差异代谢物。结果光镜下可见，对照组大鼠关节软骨细胞排列整齐，细胞形态正常；模型组大鼠膝关节软骨细胞出现大面积的变性、坏死和缺失。在两组大鼠尿液代谢物谱中，发现5种差异代谢物：4-羟基壬烯酸（hydroxynonenal，HNE）、反式．4，5．环氧基．E-2-葵稀醛（trans-4，5-epoxy．2（E）．decenal，EDE）、5-甲基脱氧胞嘧啶、5-L-谷酰基甘氨酸、脯氨酰缬氨酸。与对照组（质谱峰面积分别为65820±5200、22080±3538、4292±3520、3277±2025、1104±990）相比，模型组大鼠尿液中5种代谢物（质谱峰面积分别为90240±18863、25610±5071、9702±6562、6029±3905、4144±5322）的含量均显著升高（t=-3．903、-2．209、-2．814、-2．424、-2．174，P均〈0．05）。结论T-2毒素导致的大鼠关节软骨损伤能够引起大鼠尿液相关代谢物谱发生改变，所发现的5种差异代谢物均与软骨损伤有关。Objective To search for the metabolites that associated with articular cartilage damage in the urine of rat model with articular cartilage destruction induced by T-2 toxin. Methods Thirty healthy male Wistar rats aged 4 - 5 weeks were numbered by weight, randomly divided into two groups （n = 15 per group）, namely the control group and the model group. The rats in the control group were fed with standard rat diets, and those in the model group were given diets contaminated by T-2-toxin （300 μg/kg）. Throughout the experiment, all animals were given free access to distilled water and diets. After continuous treatment for 3 months, all the rats were sacrificed. The changes of articular cartilage in rat knee joints were observed by histopathological method, the metabolic profile of rats＂ urine was determined by ultra performance liquid chromatography/quadrupole time of flight-mass spectrometry （UPLC/QTOF-MS） technique. Combined with multivariate statistical analysis, database searching was applied to explore and confirm the different metabolites associated with cartilage damage. Results Light microscope showed that rats＂ articular chondrocytes in the control group presented ceils in neat rows and eumorphism, rats＂ articular chondrocytes in the model group presented extensive areas of chondrocyte degeneration, necrosis and loss. In rats＂ urine metabolic profiles, 5 different metabolites associated with cartilage destruction were detected, such as 4-hydroxynonenal （HNE）, trans-4,5-epoxy-2（E）-decenal （EDE）, 5-methyldeoxycytidine, and 5- L-glutamyl-glycine and prolyl-valine. Compared with the control group （mass spectrum peak area： 65 820± 5 200, 22 080 ± 3 538, 4 292±3 520, 3 277 ± 2 025, 1 104 ± 990）, all of them increased in the model group （mass spectrum peak area： 90 240± 18 863, 25 610 ± 5 071, 9 702± 6 562, 6 029 ± 3 905, 4 144 ± 5 322, t = - 3.903, - 2.209, - 2.814, - 2.424, - 2.174, all P 〈 0.05）. Conclusions The articular cartilage destruction induced by

关 键 词：T-2毒素 关节软骨 损伤 代谢组学 

分 类 号：R684[医药卫生—骨科学]