影响髓核细胞表达水通道蛋白1相关因素的实验研究  

Experimental research on the possible factors affecting the expression of aquaporin 1 in nucleus pulposus cells

作　　者：王锋[1] 朱悦[1] 

机构地区：[1]中国医科大学附属第一医院骨科,沈阳110001

出　　处：《中华临床医师杂志（电子版）》2011年第6期1634-1638,共5页Chinese Journal of Clinicians(Electronic Edition)

摘　　要：目的通过观察不同细胞外环境下,体外培养的兔髓核细胞调节水通道蛋白1(AQP-1)表达的变化,探讨AQP-1可能对髓核细胞在适应外环境变化时发挥的作用。方法体外原代培养成年兔腰椎髓核细胞成功后,在不同渗透压和氧分压条件下继续培养细胞,观察细胞的活性,并通过免疫组化和实时定量PCR观察关键蛋白aggrecan,2型胶原和AQP-1的表达变化。结果体外培养髓核细胞4～5周后可以传代,传代细胞繁殖增快,在不同的培养条件下,细胞活性无明显差异,在基因水平关键蛋白aggrecan和2型胶原表达均下降。在低渗透压条件下,细胞显著下调AQP-1表达,在低氧分压条件下,细胞显著上调AQP-1表达,在低氧分压和低渗透压条件下,AQP-1表达呈上升趋势。结论不同外环境对髓核细胞的影响不同,髓核细胞对AQP-1的表达有差异,提示AQP-1可能易化髓核细胞适应外环境变化。Objective To observe the expressions of aquaporin 1(AQP-1) in cultured nucleus pulposus cells under different osmotic stress and O2 concentration to study the possible functions of AQP-1 for nucleus pulposus cells to adapt to the changes of extracellular environment.Methods After the incubation of the rabbit lumbar nucleus pulposus cells,the cells were subjected into different culture situations with different osmotic stresses or/and different O2 concentrations.The cell viabilities were counted and the expressions of AQP-1 were observed by immunohistochemistry and real-time PCR,as well as the expressions of aggrecan and type 2 collagen with real-time PCR.Results After the primary culture for 4 to 5 weeks,the nucleus pulposus cells were expanded by two passages.When the cells were subjected into different culture situations,the cells viability were of no differences and the key proteins-aggrecan and type 2 collagen were significantly down-regulated.The nucleus pulposus cells significantly down-regulated the expression of AQP-1 in decreased osmotic stress,which were up-regulated in decreased O2 concentration.However in a decreased osmotic stress and O2 concentraiton situation,the expression of AQP-1 was of a up-trend with no significant difference.Conclusions Different extracellular environment factors could affected the rabbit nucleus pulposus cells and caused the cells to regulate the expressions of AQP-1 differently.Based upon the important functions of AQP-1,it is suggested that AQP-1 might facilitate the adaptability of nucleus pulposus cells to the changes of extracellular environment.

关 键 词：水通道蛋白质1 椎间盘 动物实验 髓核细胞 细胞外环境 

分 类 号：R681.53[医药卫生—骨科学]