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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。
Objective To investigate the effects of advanced glycation end products (AGEs) on the biological characteristics of human periodontal ligament stem cells (HPDLSCs). METHODS HPDLSCs were cultured, purified and identified. The proliferation, clonality and osteogenic differentiation of HPDLSCs were detected under different concentrations of AGEs (50,100 and 200μg / ml). Real-time PCR was used to detect the effects of different concentrations of AGEs on the expression of interleukin-1β, interleukin-6 (IL-6) and tumor necrosis factor (TNF-α) m RNA in HPDLSCs. Results MTT results showed that compared with the normal group, different concentrations of AGEs inhibited the proliferation of HPDLSCs, and 200μg / ml AGEs inhibited the most obvious. Clone results showed that compared with the normal group, different concentrations of AGEs inhibit HPDLSCs self-renewal and cloning ability, and 200μg / ml AGEs most obvious inhibition. Osteogenic differentiation ability results showed that compared with the normal group, different concentrations of AGEs inhibited HPDLSCs osteogenic ability, and 200μg / ml AGEs most obvious inhibition. The results of RT-PCR showed that compared with the normal group, different concentrations of AGEs promoted the expression of IL-1β, IL-6 and TNF-αmRNA, and with the increase of mass concentration, the expression of inflammatory cytokines increased. Conclusion AGEs inhibit the proliferation, self-renewal, clonality and osteogenic differentiation of HPDLSCs in a concentration-dependent manner and stimulate the expression of inflammatory cytokines at the same time. Therefore, the treatment of periodontal ligament stem cells for periodontal disease caused by periodontal disease with diabetes mellitus should consider the effect of advanced glycation end products.