瞬时感受电位M8在人类膝关节软骨组织中的表达  

Expression of transient receptor potential M8 in human knee joint cartilage

作　　者：喻斌[1] 聂博渊 杨朝晖[1] 

机构地区：[1]山西医科大学第二医院骨科骨与软组织损伤修复山西省重点实验室,山西省太原市030001

出　　处：《中国组织工程研究》2014年第20期3117-3121,共5页Chinese Journal of Tissue Engineering Research

基　　金：山西省留学人员科研资助项目(2013-114)~~

摘　　要：背景:瞬时感受电位通道在人体遍布多种组织和细胞,并参与人体几乎所有的生理功能和许多的病理变化。但是到目前为止,瞬时感受电位M8在人类软骨组织中的表达未见报道。目的:观察瞬时感受电位基因亚家族成员瞬时感受电位M8在人类软骨组织中的表达。方法:15例膝关节软骨样本来源于严重创伤截肢的患者。每个样本取300 mg软骨组织用于组织总蛋白的提取,再取相同部位的软骨组织做切片。Western blot方法检测瞬时感受电位M8在人类软骨组织中的表达,免疫荧光法检测瞬时感受电位M8在人类软骨组织中的分布。结果与结论:Western blot结果显示在相对分子质量为130 000时出现特异性的瞬时感受电位M8条带,并应用Westen印迹技术分析了温度敏感性瞬时感受电位M8通道的年龄相关性,发现随着年龄的增加,其表达量呈下降趋势。免疫荧光染色结果显示瞬时感受电位M8蛋白均匀的分布在细胞膜上。提示瞬时感受电位M8在人类软骨组织中有表达,具体表达在软骨细胞膜上。BACKGROUND：Transient receptor potential protein channel is distributed in many kinds of tissue and cells in human body, and takes part in many physiologic functions and pathological changes of human body. However, up to now, transient receptor potential melastatin 8 （TRPM8） has not been reported in human cartilage. OBJECTIVE：To study the expression of TRPM8, a subfamily member of transient receptor potential gene, in human knee joint cartilage. METHODS：Fifteen samples were selected from patients after amputatioin due to severe trauma. Total protein was extracted from 300 mg cartilage tissue of every patient. To detect the distribution of TRPM8 in human cartilage, cartilage sections were col ected in the same region. The expression and distribution of TRPM8 in human cartilage was detected by western blot analysis and immunofluorescence respectively. RESULTS AND CONCLUSION：Western blot analysis showed that specific TRPM8 band was visible at molecular weight of 130 000, and as the age increased, TRPM8 expression in human cartilage was gradual y decreased. Immunofluorescence staining results showed that TRPM8 was evenly located in the cellmembrane of chondrocytes. TRPM8 is expressed in the human knee cartilage, specifical y in member of chondrocytes.

关 键 词：组织构建 软骨组织工程 瞬时感受电位 软骨组织 膝关节 WESTERNBLOT 免疫荧光染色 

分 类 号：R318[医药卫生—生物医学工程]