糖基化终末产物对脂多糖介导下人牙周膜干细胞增殖和骨向分化能力的影响

Effect of advanced glycation end products on proliferation and osteogenic differentiation of lipopolysaccharide-stimulated human periodontal ligament stem cells

目的:探讨糖基化终末产物(AGEs)对脂多糖(LPS)介导下的人牙周膜干细胞(HPDLSCs)增殖和骨向分化能力的影响。方法:HPDLSCs培养和鉴定;MTT法检测不同浓度LPS和AGEs对HPDLSCs增殖能力的影响;Real Time-PCR检测10μg/mL LPS和100μg/mL LAGE_S刺激HPDLSCs后白细胞介素-6(IL-6)、碱性磷酸酶(ALP)和Runt-related transcription factor2(RUNX-2)mRNA的表达水平。结果:与正常组比较,10、100μg/mL LPS均抑制HPDLSCs的增殖,50、100、200μg/mL AGEs均抑制HPDLSCs的增殖,100、200μg/mL AGEs比50μg/mL AGEs抑制明显,差异有统计学意义(P<0.05);在成骨诱导下10μg/mL LPS刺激HPDLSCs 6 d和10μg/mL LPS刺激HPDLSCs 3d后再加10μg/mL LPS/100μg/mL AGES共同刺激3d,后者较前者IL-6表达明显增加,ALP和RUNX-2的表达降低,差异有统计学意义(P<0.05)。结论:LPS和AGEs均成浓度依赖性抑制HPDLSCs的增殖能力。AGEs能使LPS介导下的HPDLSCs的炎性因子表达增强,成骨分化能力降低。提示糖尿病可能会加重牙周炎病人的牙周炎炎症程度。

AIM：To investigate the effect of advanced glycation end products（AGEs） on proliferation and osteogenic differentiation of lipopolysaccharide（LPS）-stimulated human periodontal ligament stem cells（HPDLSCs）. METHODS：The effects of LPS and AGEs on the proliferation of HPDLSCs were analyzed by MTT assay.The expressions of interleukin-6（IL-6）,alkaline phosphatase（ALP） and Runt-related transcription factor-2（RUNX-2） mRNA of HPDLSCs which were stimulated by LPS（10μg/mL） and AGEs（100μg/mL） were detected by RT-PCR.RESULTS： MTT assay showed that LPS and AGEs inhibited the proliferation of HPDLSCs in a dose dependent manner. LPS stimulation significantly increased IL-6 expression and inhibited ALP and RUNX-2 expression in HPDLSCs.Co -stimulation of HPDLSCs with LPS and AGEs markedly enhanced the effects of LPS on IL-6,ALP and RUNX-2 expression. CONCLUSION：LPS and AGEs inhibited the proliferation of HPDLSCs in a concentration-dependent man- ner.AGEs can enhance the stimulatory effects of LPS in up-regulating IL-6 expression and the inhibitory effects of LPS in down-regulating osteogenic factors expression.Our results suggest that diabetes may increase the degree of periodontal inflammation in patients with periodontitis.