CAJ | 학술논문

目的比较大鼠骨髓间充质干细胞(BMSCs)成骨分化过程中神经肽Y(neuropeptide Y,NPY)Y1受体的表达变化.方法采用全骨髓法体外分离、培养SD大鼠BMSCs,分为诱导成骨组与未诱导组,在传代培养的不同时期(1,2,3周),采用免疫细胞化学、Western blot对BMSCs的成骨细胞诱导进行鉴定,采用荧光定量RT-PCR、Western blot检测各组细胞神经肽Y Y1受体mRNA、蛋白的表达情况.结果 RT-PCR结果显示,同一时相点诱导组Y1受体mRNA表达量低于未诱导组,诱导组和未诱导组Y1受体mRNA表达均以时间依赖性的方式增高.Western blot 结果显示,同一时相点诱导组Y1受体的蛋白水平高于未诱导组,诱导组和未诱导组Y1受体蛋白水平以时间依赖性的方式减少.结论大鼠BMSCs向成骨细胞诱导分化过程中,神经肽Y Y1受体mRNA及其蛋白表达呈现不同趋势,神经肽Y可能通过Y1受体通路介导对BMSCs成骨分化的抑制作用.
목적 비교대서골수간충질간세포(BMSCs)성골분화과정중신경태Y(neuropeptide Y,NPY)Y1수체적표체변화.방법 채용전골수법체외분리、배양SD대서BMSCs,분위유도성골조여미유도조,재전대배양적불동시기(1,2,3주),채용면역세포화학、Western blot대BMSCs적성골세포유도진행감정,채용형광정량RT-PCR、Western blot검측각조세포신경태Y Y1수체mRNA、단백적표체정황.결과 RT-PCR결과현시,동일시상점유도조Y1수체mRNA표체량저우미유도조,유도조화미유도조Y1수체mRNA표체균이시간의뢰성적방식증고.Western blot 결과현시,동일시상점유도조Y1수체적단백수평고우미유도조,유도조화미유도조Y1수체단백수평이시간의뢰성적방식감소.결론 대서BMSCs향성골세포유도분화과정중,신경태Y Y1수체mRNA급기단백표체정현불동추세,신경태Y가능통과Y1수체통로개도대BMSCs성골분화적억제작용.
Objective To compare the expression changes of neuropeptide Y (NPY) receptor Y1 in different stages of osteoblast differentiation of rat bone marrow mesenchymal stem cells (BMSCs).Methods The rBMSCs were isolated in vitro from Sprague-Dawley (SD) rats using whole bone marrow adherence method and cultured. Then, the rBMSCs were divided into osteoblast-induced group and noninduced group. In different periods of culture at 1, 2 and 3 weeks, identification of the osteoblasts was performed by using immunocytochemistry and Western blot. Expressions of mRNA and protein of Y1 receptor were detected by real time reserve transcriptase-polymerase chain reaction (RT-PCR) and Western blot. Results RT-PCR demonstrated that osteoblast-induced group had a lower expression of Y1 receptor than non-induced group at the same time point and the expression of Y1 receptor was increased in a time-dependent manner in both groups. Western blot demonstrated higher expression of Y1 receptor in osteoblast-induced group compared with non-induced group at the same time point and a decreased expression of Y1 receptor in a time-dependent manner in both groups. Conclusions During the process of osteoblastic differentiation of rat BMSCs, the expressions of mRNA and protein of NPY Y1 receptor show different trends, when NPY may mediate the inhibition of osteoblastic differentiation of BMSCs through Y1 receptor pathway.