牙周膜干细胞中乙酰基转移酶MORF调控成骨的研究

目的：：比较正常与炎症来源牙周膜干细胞( H-PDLSCs与P-PDLSCs)中乙酰基转移酶MORF表达水平的差异及其对PDLSCs分化的调控。方法：有限稀释法培养H-PDLSCs与P-PDLSCs, LPS、 TNF-α、IL-β及三者混合物体外模拟炎症微环境诱导H-PDLSCs ( IP-PDLSCs )；基因与蛋白检测方法对比2种来源 PDLSCs 中 MORF 的表达水平；蛋白检测方法检测 IP-PDLSCs中MORF的表达水平；基因、蛋白检测与茜素红染色方法对比H-PDLSCs及下调MORF基因后的PDLSCs成骨基因表达的差异。结果：与 H-PDLSCs 相比,P-PDLSCs 中 MORF 的表达显著下降(P <0.05)；IP-PDLSCs 中 MORF 的表达下降；MORF基因被下调后,PDLSCs成骨能力下降(P<0.05)。结论：牙周炎及炎症微环境会导致PDLSCs中MORF表达的下降,从而使其成骨分化受到抑制。

Acetyltransferase MORF regulates osteogenic differentiation potential of periodontal ligament stem cells

Objective:To compare acetyltransferase MORF level in periodontal ligament stem cells( PDLSCs) derived from healthy individuals ( H-PDLSCs) with those derived from the individuals with periodontitis ( P-PDLSCs ) . And to determine the effect of MORF on the osteogenic differentiation potential of PDLSCs. Methods: Human H-PDLSCs and P-PDLSCs were cultured and cloned with limited dilution method. H-PDLSCs were stimulated by LPS, TNF-α, IL-β and the mix of the 3 inflammatory factors to imitate inflammatory environment ( IP-PDLSCs ) . Quantitative RT-PCR and Western Blot were applied to examine different expression of MORF in H-PDLSCs and P-PDLSCs. Western Blot was applied to detect expression of MORF in IP-PDLSCs. Quantitative RT-PCR, Western Blot and alizarin red staining were applied to determine osteogenic differentiation potential of H-PDLSCs with MORF knock-down. Results:Quantitative RT-PCR and Western Blot showed lower expression of MORF in P-PDLSCs compared with H-PDLSCs( P<0. 05). Western Blot revealed lower expression of MORF in IP-PDLSCs. Quantitative RT-PCR, Western Blot and alizarin red stai-ning indicated osteogenic differentiation potential was inhibited in H-PDLSCs with MORF knockdown(P<0. 05). Conclusion: Peri-odontitis can suppress the expression of MORF in PDLSCs and inhibite the osteogenic differentiation potential of PDLSCs.