CAJ | 학술논문

目的通过检测前列腺素E2( prostaglandins E2，PGE2)作用于MC3T3 - E1成骨细胞后基因表达谱的改变，探索PGE2促进骨形成作用的分子机制。方法 10 μmol/L的PGE2处理MC3T3 - E1成骨细胞30 min后，用基因芯片技术检测PGE2处理成骨细胞后基因表达谱的变化，选取在骨再生过程中重要的基因用Westem blot法检测验证。结果 PGE2处理成骨细胞后，276个基因表达上调，其中和骨再生有关的基因主要有单核细胞向巨噬细胞转化基因( monocyte to macrophage differentiation，MMD)、核受体基因(nuclear receptor subfamily 4，group A，member 2，NR4A2)、骨形态发生蛋白-7（bone morphogenetic protein-7，BMP -7）、成骨细胞特异性因子(periostin，osteoblast specific factor，POSTN)和钙黏蛋白等；168个基因表达下调，其中和骨再生有关的基因有DNA结合抑制因子1，2，3(Id1，2，3)等。Western blot结果显示，与对照组比较，PGE2处理成骨细胞后核因子- Κb( nuclear factor - Κb，NF - Κb) p65和BMP -7蛋白表达显著上升(P＜0.01)，Id2蛋白表达显著下降(P＜0.O1)，与基因芯片结果基本一致。结论结合基因芯片结果和Westem blot结果，可以推测PGE2处理成骨细胞后首先激活核受体基因NR4A2，继而引起NF - Κb的活化，最后引起下游基因BMP -7和Id2等的变化从而引起成骨细胞分化，促进骨再生。
목적 통과검측전렬선소E2( prostaglandins E2，PGE2)작용우MC3T3 - E1성골세포후기인표체보적개변，탐색PGE2촉진골형성작용적분자궤제。방법 10 μmol/L적PGE2처리MC3T3 - E1성골세포30 min후，용기인심편기술검측PGE2처리성골세포후기인표체보적변화，선취재골재생과정중중요적기인용Westem blot법검측험증。결과 PGE2처리성골세포후，276개기인표체상조，기중화골재생유관적기인주요유단핵세포향거서세포전화기인( monocyte to macrophage differentiation，MMD)、핵수체기인(nuclear receptor subfamily 4，group A，member 2，NR4A2)、골형태발생단백-7（bone morphogenetic protein-7，BMP -7）、성골세포특이성인자(periostin，osteoblast specific factor，POSTN)화개점단백등；168개기인표체하조，기중화골재생유관적기인유DNA결합억제인자1，2，3(Id1，2，3)등。Western blot결과현시，여대조조비교，PGE2처리성골세포후핵인자- Κb( nuclear factor - Κb，NF - Κb) p65화BMP -7단백표체현저상승(P＜0.01)，Id2단백표체현저하강(P＜0.O1)，여기인심편결과기본일치。결론 결합기인심편결과화Westem blot결과，가이추측PGE2처리성골세포후수선격활핵수체기인NR4A2，계이인기NF - Κb적활화，최후인기하유기인BMP -7화Id2등적변화종이인기성골세포분화，촉진골재생。
Objective To investigate the molecular mechanism of prostaglandins E2 ( PGE2 ) in promoting bone formation by detecting the changes of gene expression profiles of MC3T3-E1 osteoblasts treated with PGE2. Methods The genes with differential expression in MC3T3-E1 osteoblasts treated with 10 μmol/L PGE2 for 30 minutes were performed by gene chip technology. Several major genes during bone regeneration were selected for Western blot analysis. Results After co-culture of MC3T3-E1 cells with PGE2 at concentration of 10 μmol/L for 30 min, 276 genes were up-regulated, including bone regeneration related MMD (monocyte to macrophage differentiation associated), NR4A2 (nuclear receptor subfamily 4, group A, member 2), BMP-7 ( bone morphogenetic protein-7), POSTN ( periostin, osteoblast specific factor) and catenin (cadherin-associated protein) genes; and 168 genes were down-regulated,including bone regeneration related Idl ,2,3 ( inhibitor of DNA binding 1,2,3 ) genes. Western blot analysis indicated that the expressions of nuclear factor (NF)-κB p65 and BMP-7 protein in the osteoblasts treated with 10 μmol/l PGE2 were apparently higher ( P ＜ 0. Ol ) than that of the controls, whereas the ld2 expression decreased (P ＜0. O1 ) under the same conditions, which was almost the same as the results of gene chip technology. Conclusions With the results of gene chip and Western blot, it can be speculated that the PGE 2 firstly activates the nuclear receptor NR4A2 and then the nuclear transcription factor NF-κB, induces the changes of the downstream gene BMP-7 and Id2 expression and finally results in the differentiation of the osteoblasts and promote the bone regeneration.