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目的:探讨牙髓卟啉单胞菌(Porphyromonas endodontalis,P.e)脂多糖(lipopolysaccharide,LPS)对成骨细胞产生巨噬细胞炎性蛋白1α(macrophageinflammatoryprotein-1α,MIP-1α)m RNA和蛋白分泌的影响,以及姜黄素对此过程产生的抑制作用。方法:以20 mg/L P.e-LPS作用细胞不同时间(0~48 h)后,采用实时反转录聚合酶链反应(real-time reverse transcription-polymerase chain reaction,real-time RT-PCR)和酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测MIP-1αm RNA和蛋白的表达。以同样方法检测姜黄素对20 mg/L P.e-LPS刺激MC3T3-El细胞后MIP-1αm RNA和蛋白表达的影响。采用SPSS 13.0软件包对结果进行单因素方差分析和Dunnett t检验。结果:在观察时间内(0~48 h),20 mg/L P.e-LPS作用MC3T3-El细胞后,MIP-1αm RNA的表达和蛋白的分泌具有时间依赖性。10 mol/L姜黄素预处理细胞1 h,可以降低P.e-LPS诱导的MIP-1αm RNA和蛋白的表达水平。结论:P.e-LPS可以诱导成骨细胞表达和分泌MIP-1α,姜黄素对此过程发挥明显的抑制作用。
Objective: To investigate the effects of Porphyromonas endodontalis (Pe) lipopolysaccharide (LPS) on osteoblast-derived macrophage inflammatory protein-1α (MIP-1α) m RNA and protein secretion Effect, as well as curcumin on the inhibition of this process. Methods: Real-time reverse transcription-polymerase chain reaction (real-time RT-PCR) and real-time reverse transcription-polymerase chain reaction Enzyme linked immunosorbent assay (ELISA) was used to detect the expression of MIP-1α mRNA and protein. In the same way, the effect of curcumin on MIP-1αm RNA and protein expression in MC3T3-El cells stimulated by 20 mg / L P.e-LPS was measured. One-way ANOVA and Dunnett t-test were used to analyze the results using SPSS 13.0 software package. Results: The expression of MIP-1αmRNA and the secretion of protein in MC3T3-El cells treated with 20 mg / L P.e-LPS in a time-dependent manner (0-48 h) were time-dependent. Pretreatment with 10 mol / L curcumin for 1 h decreased the expression of MIP-1αmRNA and protein induced by P.e-LPS. CONCLUSION: P.e-LPS can induce osteoblasts to express and secrete MIP-1α, and curcumin exerts a significant inhibitory effect on this process.