Inhibitory effects of advanced glycation end-products and Porphyromonas gingivalis lipopolysaccharide on the expression of osteoblastic markers of rat bone marrow cells in culture
Inhibitory effects of advanced glycation end-products and Porphyromonas gingivalis lipopolysaccharide on the expression of osteoblastic markers of rat bone marrow cells in culture
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DOI：
10.1111/jre.12310
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发表时间：
2016-06-01
影响因子：
3.5
通讯作者：
Nagata, T.
中科院分区：
文献类型：
--
作者：
Sakamoto, E.;Mihara, C.;Nagata, T.
背景和目标：糖尿病是牙周炎的主要危险因素，高血糖的程度与牙周炎的严重程度密切相关。晚期糖基化终产物（AGEs）在糖尿病条件下在各种组织中积累。牙周组织中的AGEs可能在上调牙周炎症中发挥作用;然而，AGEs与牙周炎严重程度的关系尚未完全阐明。牙龈卟啉单胞菌脂多糖（P-LPS）是牙周炎的重要致病因子。虽然AGE或P-LPS对成骨细胞的独立作用已在体外报道，但两者的加入效果尚未明确阐明。在本研究中，探讨加重糖尿病牙周炎的因素，我们研究了AGE和P-LPS对成骨细胞标志物的表达和炎症相关标志物的表达的影响，invitro.Material和方法：大鼠骨髓细胞培养，碱性磷酸酶活性和骨结节形成作为成骨细胞标志物进行了评估。进行逆转录聚合酶链反应以确定与骨和炎症相关的分子的mRNA表达。骨钙素和白细胞介素-1 β（IL-1 β）的蛋白水平测定使用酶联免疫吸附试验。结果：AGEs和P-LPS独立地降低碱性磷酸酶活性和骨结节形成。加入AGE和P-LPS（AGE+P-LPS）进一步降低了这些标志物。逆转录-聚合酶链反应分析显示，AGE+P-LPS显著降低成骨细胞相关分子如1型胶原、骨钙素和Cbfa 1的mRNA表达，显著增加炎症相关分子如IL 1 β和S100 A8的mRNA表达。结论：AGEs和P-LPS均能抑制大鼠骨髓成骨细胞标志物的表达，增加炎症标志物的水平，提示AGEs和P-LPS可能是糖尿病牙周炎加重的重要因素。
Background and Objectives: Diabetes is a major risk factor for periodontitis and there is a close relationship between the degree of hyperglycemia and the severity of periodontitis. Advanced glycation end-products (AGEs) accumulate in various tissues under diabetic conditions. AGEs in the periodontal tissues probably play a role in upregulating periodontal inflammation; however, the association of AGEs with the severity of periodontitis has not been fully clarified. Lipopolysaccharide from Porphyromonas gingivalis (P-LPS) is a potent pathogenic factor in periodontitis. Although the independent effect of AGE or P-LPS on osteoblastic cells has been reported invitro, the effect of adding both has not been clearly elucidated. In this study, to explore factors aggravating diabetic periodontitis, we investigated the effects of AGE and P-LPS on the expression of osteoblastic markers and the expression of inflammation-related markers invitro.Material and Methods: Rat bone marrow cells were cultured, and alkaline phosphatase activity and bone nodule formation were evaluated as osteoblastic markers. Reverse transcription-polymerase chain reaction was performed to determine the mRNA expression of molecules associated with bone and inflammation. Protein levels of osteocalcin and interleukin-1 beta (IL-1 beta) were measured using enzyme-linked immunosorbent assay.Results: AGEs and P-LPS independently reduced alkaline phosphatase activity and bone nodule formation. The addition of both AGE and P-LPS (AGE+P-LPS) further decreased these markers. Reverse transcription-polymerase chain reaction analysis revealed that AGE+P-LPS markedly decreased the mRNA expression of osteoblast-related molecules such as type 1 collagen, osteocalcin and Cbfa1, and markedly increased that of inflammation-related molecules such as IL1 beta and S100A8. AGE and P-LPS decreased the protein level of osteocalcin and increased that of IL-1 beta, and a further increase of IL-1 beta was detected for AGE+P-LPS.Conclusion: AGEs and P-LPS inhibited the expression of osteoblastic markers and increased the levels of inflammatory markers in rat bone marrow cells, suggesting that both AGE and P-LPS may be important factors associated with the aggravation of diabetic periodontitis.