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PPARγ配体15d-PGJ2(15-deoxy-Δ12,14-prostaglandin J2)对细胞有抑制增殖、促进分化和凋亡的作用,但是它对骨髓间充质干细胞(BM-MSC)培养上清中细胞因子的影响还未见报道。本研究探讨15d-PGJ2对BM-MSC培养上清中细胞因子的影响。首先对正常人骨髓进行分离培养获得可传代的贴壁生长的成纤维细胞样细胞,然后应用流式细胞术检测细胞的表型,应用条件培养液诱导细胞向成脂和成骨分化以鉴定所获得细胞为BM-MSC。在BM-MSC培养体系中加入10、20、40和60μmol/L的15d-PGJ2并培养24 h,用实时定量PCR测定PPARγmRNA水平,共聚焦免疫荧光技术检测PPARγ的表达水平。结果发现,当15d-PGJ2的浓度达到20μmol/L时细胞出现了凋亡并大量从培养瓶表面脱落;而10μmol/L的15d-PGJ2刺激细胞24 h后PPARγ的mRNA表达水平增高,与对照组相比有显著性差异(P<0.05)。然后用含有10μmol/L 15d-PGJ2和不含15d-PGJ2体系分别培养BM-MSC 24 h并用蛋白芯片检测培养上清,发现有部分因子表达水平发生了变化。结论:TIMP-2在15d-PGJ2刺激后下调,且信号值及变化水平有意义。
PPARγ ligand 15d-PGJ2 (15-deoxy-Δ12, 14-prostaglandin J2) can inhibit proliferation, promote differentiation and apoptosis of cells, but it can inhibit the proliferation of bone marrow mesenchymal stem cells The impact of factors has not been reported. This study was to investigate the effect of 15d-PGJ2 on cytokines in BM-MSC culture supernatant. First of all, the normal human bone marrow was isolated and cultured to obtain adherent adherent growth of fibroblast-like cells, and then the use of flow cytometry to detect the cell phenotype, the conditioned medium was used to induce cells to differentiate into adipogenic and osteogenic to identify The cells obtained were BM-MSCs. 15d-PGJ2 at 10, 20, 40 and 60μmol / L was added into BM-MSC culture system and cultured for 24 hours. The level of PPARγ mRNA was detected by real-time quantitative PCR and the expression of PPARγ was detected by confocal immunofluorescence. The results showed that when the concentration of 15d-PGJ2 reached 20μmol / L, the cells appeared apoptosis and a large number of detached from the flask surface; 15μmol / L 15d-PGJ2 stimulated cells 24 h after PPARγ mRNA expression increased, compared with the control group Compared with significant difference (P <0.05). Then, BM-MSCs were cultured for 24 h in 10μmol / L 15d-PGJ2 and 15d-PGJ2 respectively, and the supernatant was detected by protein microarray. Some of the factors were found to be changed. Conclusion: TIMP-2 is down-regulated after 15d-PGJ2 stimulation, and the signal value and the level of change are significant.