CAJ | 학술논문

目的研究神经递质P物质调控成骨细胞分化的分子途径.方法分离骨髓基质干细胞进行原代及传代培养;分别采用空白对照、P物质、P物质NK1受体拮抗剂、P物质+P物质NK1受体拮抗剂进行干预,诱导骨髓基质干细胞向成骨细胞分化;传代培养1～2周后,抽提细胞总RNA,用RT-PCR检测分化过程中Osterix基因的表达.检测结果重复3次,采用单因素方差分析检测结果.结果骨髓基质干细胞在生长对数增殖期为4～6 d,采用RT-PCR检测发现P物质干预成骨细胞分化,导致成骨细胞分化过程中重要的转录引子Osterix 基因表达,与其他各组比较有显著差异(P<0.05),Osterix基因表达上调,从而刺激前成骨细胞向成骨细胞转化.而P物质+P物质NK1受体拮抗剂共同干预,Osterix基因表达与空白对照组无显著差异(P<0.05),说明P物质通过P物质NK1受体对成骨细胞分化进行调控.结论 P物质可调控前成骨细胞分化过程中转录因子Osterix基因表达促进其向成骨细胞分化.P物质对Osterix基因表达的调控依赖P物质NK1受体.
목적 연구신경체질P물질조공성골세포분화적분자도경.방법 분리골수기질간세포진행원대급전대배양;분별채용공백대조、P물질、P물질NK1수체길항제、P물질+P물질NK1수체길항제진행간예,유도골수기질간세포향성골세포분화;전대배양1～2주후,추제세포총RNA,용RT-PCR검측분화과정중Osterix기인적표체.검측결과중복3차,채용단인소방차분석검측결과.결과 골수기질간세포재생장대수증식기위4～6 d,채용RT-PCR검측발현P물질간예성골세포분화,도치성골세포분화과정중중요적전록인자Osterix 기인표체,여기타각조비교유현저차이(P<0.05),Osterix기인표체상조,종이자격전성골세포향성골세포전화.이P물질+P물질NK1수체길항제공동간예,Osterix기인표체여공백대조조무현저차이(P<0.05),설명P물질통과P물질NK1수체대성골세포분화진행조공.결론 P물질가조공전성골세포분화과정중전록인자Osterix기인표체촉진기향성골세포분화.P물질대Osterix기인표체적조공의뢰P물질NK1수체.
Objective To study the molecular pathway of osteoblastic differentiation induced by substance P (SP), a neurotransmitter. Methods Mesenchymal stem cells were isolated and cultured, and treated with SP or its receptor (NK1) antagonist to induce osteoblastic cell differentiation, respectively. Alkaline phosphatase activity was determined; Osterix gene expression was detected by RT-PCR after 1-2 weeks for three times. The data of each culture condition were analyzed using SPSS12.0 statistical software to determine whether the differences between conditions were significant. Results After 4-5 days' culture, bone marrow stromal cells became spindle-shaped, triangular or polygonic. They covered the plate surface, formed extensive cell sheets in each group after 11-12 days of culture, and then induced differentiation to osteoblast. SP up-regulated the important transcription factor Osterix gene expression significantly (P<0.05). Conclusion The up-regulation of Osterix gene expression by SP may stimulate osteoblastic cell differentiation. SP's regulation depends on its receptor NK1.