葡萄糖转运子9和尿酸转运子1在软骨细胞中表达的体外试验  

Expression of glucose transporter 9 and urate transporter 1 in chondrocytes in vitro

作　　者：张冰清[1] 段梦园 张保中[3] 龙波 王冬梅[5] 朱蕾 曾学军[6] ZHANG Bing-qing;DUAN Meng-yuan;ZHANG Bao-zhong;LONG Bo;WANG Dong-mei;ZHU Lei;ZENG Xue-jun(Department of Internal Medicine, Peking Union Medical College Hospital, Chinese Academy of Medical Science, Beijing 100730, China)

机构地区：[1]中国医学科学院北京协和医院内科,北京100730 [2]中国医学科学院基础学院药理学系,北京100010 [3]中国医学科学院北京协和医院骨科,北京100730 [4]中国医学科学院北京协和医院中心实验室,北京100730 [5]南方医科大学南方医院神经内科,广东广州510000 [6]中国医学科学院北京协和医院普通内科,北京100730

出　　处：《中国临床研究》2019年第7期870-873,共4页Chinese Journal of Clinical Research

基　　金：中国医学科学院医学与健康科技创新工程项目(2016-I2M-1-002);中国医学科学院医学表观中心基金项目(2017PT31035);北京协和医院青年科学基金项目(pumch-2016-2.13)~~

摘　　要：目的观察在软骨细胞中葡萄糖转运子(GLUT)9和尿酸转运子(URAT)1的表达,探讨软骨细胞中是否存在尿酸转运。方法选择2017年11月至2018年4月5例外伤性股骨颈骨折患者(排除痛风性等关节炎和病理性骨折)的软骨组织作为对象,用Western blot和免疫组化法测定软骨组织中GLUT9和URAT1蛋白表达水平,并以Ⅱ型胶原蛋白作为阳性对照。结果 Western blot结果表明,5例标本Ⅱ型胶原蛋白均为阳性,Ⅱ型胶原蛋白是软骨细胞的特征性蛋白,其染色阳性证实取材标本中包含软骨细胞。5例标本在59 kDa和70 kDa均有阳性条带,分别对应兔抗人GLUT9和兔抗人URAT1;免疫组化结果表明,软骨细胞样本中可见特征性的软骨窝结构;软骨细胞经Ⅱ型胶原蛋白抗体染色后细胞质均显现为褐色,URAT1和GLUT9两种抗体染色下,软骨细胞的细胞膜可见均一的褐色染色,细胞质内亦有褐色染色分布。结论 GLUT9和URAT1在软骨组织中呈高表达,可能存在尿酸转运过程,其是否参与痛风性关节炎的发生、发展,有待进一步研究。Objective To investigate the expressions of glucose transporter 9(GLUT9) and urate transporter 1(URAT1) in chondrocytes and to explore whether uric acid transport exists in chondrocyte.Methods The cartilage tissues from 5 patients with traumatic femoral neck fracture(excluding gouty arthritis and pathological fracture) collected from November 2017 to April 2018 were selected as subjects.The expression levels of GLUT9 and URAT1 in cartilage tissue were determined by Western blot and immunohistochemistry,and type Ⅱ collagen was used as a positive control.Results Western blot results showed that type Ⅱ collagen was positive in all 5 samples.Type Ⅱ collagen was the characteristic protein of chondrocyte.Positive staining of type Ⅱ collagen confirmed that chondrocyte was contained in the samples.Five specimens presented positive bands at 59 kDa and 70 kDa,corresponding to rabbit anti-human GLUT9 and rabbit anti-human URAT1.Immunohistochemistry results showed that characteristic cartilage fossa was visible in chondrocyte samples.The cytoplasm of the chondrocytes stained with type Ⅱ collagen antibody showed brown color.By the staining of URAT1 and GLUT9 antibody,the cell membrane of chondrocytes appeared uniform brown staining,and brown staining distribution was also found in the cytoplasm of chondrocytes.Conclusions High expressions of GLUT9 and URAT1 in cartilage tissue may be involved in uric acid transport.Whether GLUT9 and URAT1 participate in the occurrence and development of gouty arthritis needs further study.

关 键 词：软骨细胞 葡萄糖转运子9 尿酸转运子1 免疫印迹法 免疫组织化学法 尿酸转运 痛风性关节炎 

分 类 号：R589.7[医药卫生—内分泌]