Notch信号通路相关分子在牙周膜干细胞中的表达及意义

Notch signaling pathway related molecules in the expression and significance of the periodontal ligament stem cells

目的:检测Notch信号通路相关分子在炎性和正常牙周膜干细胞中的表达差异,探讨其在牙周炎牙槽骨缺损中的作用机制。方法:组织块酶消化法培养正常组织及慢性牙周炎组织来源的的牙周膜干细胞(H-PDLSCs,PPDLSCs),并经有限稀释法纯化两种细胞,采用免疫荧光法检测干细胞表面标记物CD146及PDLSCs表面蛋白STRO-1的表达,通过免疫荧光染色和实时荧光定量PCR检测Notch信号通路中Notch1受体蛋白、配体Jagged1(JAG1)信号分子的表达变化。结果:两种来源的细胞经纯化后均阳性表达间充质干细胞表面标志物STRO-1、CD146;P-PDLSCs比H-PDLSCs具有更强的增殖能力;免疫荧光染色检测显示,H-PDLSCs和P-PDLSCs均阳性表达Notch1、Jagged1;但PPDLSCs中Notch1、Jagged1的阳性表达明显较弱;RT-PCR检测发现P-PDLSCs中Notch1、Jagged1m RNA表达量分别为(1.22±0.29)、(1.52±0.38),较H-PDLSCs明显降低,差异有统计学意义(均P<0.05)。结论:炎症牙周膜干细胞中的炎症微环境可能抑制了Notch信号分子的表达,低水平的Notch信号的激活可能有利于牙周炎缺损牙槽骨的成骨分化。

Objective：Detection of Notch signaling pathway related molecules in inflammatory and normal periodontal membrane expression differences of cells,and to explore its mechanism in periodontitis alveolar defect. Method：H-PDLSCs and P-PDLSCs were primarily cultured by tissue digestion method. Cells were passaged and identified by stem cell surface marker expression using immunofluorescence staining.Detection of Notch1 receptor proteins and ligand Jagged1 signal molecule expression in the Notch signal pathway by immunofluorescence test and real-time quantitative PCR. Result：The two types of cells both expressed stem cell markers STRO-1 and CD146. P-PDLSCs showed higher multiplication capacity. Both cells all positive expression of Notch1 and Jagged1;The expression of Notch1 m RNA and Jagged1 m RNA in PPDLSCs were（1.22±0.29）,（1.52±0.38）respectively,they were lower than those in H-PDLSCs,the differences were statistically significant（P〈0.05）. Conclusion：Inflammatory periodontal membrane inflammation of stem cell microenvironment may inhibit the expression of the Notch signaling molecules,the low lenel of the activation of the Notch signal may be good for periodontitis defect alveolar bone ossification of differentiation.