糖基化终末产物对人牙周膜干细胞生物学特性的影响

Effects of advanced glycation end products on the biological characteristics of periodontal ligament stem cells

目的探讨糖基化终末产物(advanced glycation end products,AGEs)对人牙周膜干细胞(human periodontal ligament stem cells,HPDLSCs)生物学特性的影响研究。方法 HPDLSCs培养、提纯以及鉴定;检测不同质量浓度AGEs(50、100、200μg/ml)下HPDLSCs增殖能力、克隆能力、成骨分化能力。Real-time PCR检测不同质量浓度AGEs对HPDLSCs白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和肿瘤坏死因子(TNF-α)m RNA表达的影响。结果 MTT结果显示,与正常组比较,不同质量浓度AGEs均抑制HPDLSCs的增殖,且200μg/ml AGEs抑制最明显。克隆结果显示,与正常组比较,不同质量浓度AGEs均抑制HPDLSCs的自我更新及克隆能力,且200μg/ml AGEs抑制最明显。成骨分化能力结果显示,与正常组比较,不同质量浓度AGEs均抑制HPDLSCs的成骨能力,且200μg/ml AGEs抑制最明显。RT-PCR结果显示,与正常组比较,不同质量浓度AGEs均促进IL-1β、IL-6和TNF-αm RNA的表达,且随着质量浓度的增加,炎性因子的表达增加。结论 AGEs成浓度依赖性抑制HPDLSCs的增殖能力、自我更新能力、克隆能力和成骨分化能力,同时刺激炎性因子的表达。因此,牙周膜干细胞治疗糖尿病伴牙周病导致的牙周组织损伤要考虑糖基化终末产物的影响。

This study designed to study the effects of advanced glycation end products（AGEs） on the biological characteristis of periodontal ligament stem cells（HPDLSCs）. HPDLSCs were cultured from normal health donors, and the ability of HPDLSCs to undergo osteogenic differentiation was detected after treatment with AGEs of different concentrations（50, 100, 200 μg/ml）. Real-time PCR was used to detect m RNA expressions of interleukin-1β（IL-1β）, interleukin-6（IL-6） and tumor necrosis factor（TNF-α） in HPDLSCs under different concentration of AGEs. MTT results showed that, compared with control group, AGEs of all concentrations were capable to inhibit HPDLSCs ability of self-renewal and cloning, and the ability of undergoing osteogenic differentiation; RT-PCR showed that the expressions of IL-1β, IL-6 and TNF-α m RNA were up-regulated with the increasing of AGEs concentration in culture. In conclusion, AGEs inhibit HPDLSCs proliferation, self-renewal, clonal and osteogenic differentiation abilities in a concentration-dependent manner. In addition, AGEs activate the expression of inflammatory cytokines. Thus the AGEs effect on the HPDLSCs needs to be considered in the treatment of diabetic periodontitis.