Asxl1在炎性微环境中对成骨细胞增殖分化的作用

背景:研究表明Asxl1的缺失可导致骨质发育不全、骨质缺损类疾病的发生,但目前在根尖周炎环境下该因子与骨破坏之间的关系暂无相关报道。目的:探讨炎性微环境下Asxl1对成骨细胞增殖分化的影响。方法:实验选用脂多糖刺激MC3T3-E1细胞建立体外炎性微环境,通过CCK-8实验筛取脂多糖最佳质量浓度和最佳作用时间,然后用20 mg/L脂多糖刺激MC3T3-E1细胞24 h,免疫荧光检测Asxl1的蛋白表达水平,Real Time-PCR检测Asxl1 mRNA的表达水平。为进一步验证Asxl1基因在炎性微环境中影响成骨细胞的增殖与分化,脂多糖刺激形成炎性微环境后转染Asxl1-SiRNA 24 h,采用CCK-8检测细胞增殖活性,RealTime-PCR检测Asxl1及成骨相关基因ALP和RUNX2 mRNA的表达水平。结果与结论:①脂多糖刺激MC3T3-E1细胞后,Asxl1蛋白和mRNA表达水平呈降低趋势;②脂多糖刺激MC3T3-E1细胞后,转染Asxl1-SiRNA 24 h,细胞增殖活性下降趋势明显,Asxl1基因及成骨相关基因ALP和RUNX2 mRNA的表达水平明显降低;③结果提示,Asxl1可能通过参与炎性反应过程,影响成骨细胞的增殖与分化,进而参与骨破坏进程。

Effect of sex combing protein 1 on proliferation and differentiation of osteoblasts in inflammatory microenvironment

BACKGROUND:Studies have shown that the loss of sex combing protein 1(Asxl1)can lead to the occurrence of bone dysplasia and bone defects,but the relationship between this factor and bone destruction in the microenvironment of apical periodontitis has not been reported.OBJECTIVE:To study the effect of Asxl1 on proliferation and differentiation of osteoblasts in an inflammatory microenvironment.METHODS:MC3T3-E1 cells were excited by lipopolysaccharide to establish an in vitro inflammatory microenvironment.The best concentration and optimal action time of lipopolysaccharide were screened by cell counting kit-8 test.MC3T3-E1 cells were then stimulated with 20 mg/L lipopolysaccharide for 24 hours.The expression levels of Asxl1 protein and mRNA were detected by immunofluorescence and real-time PCR,respectively.After lipopolysaccharide stimulated the formation of inflammatory microenvironment,Asxl1-Si RNA was transfected for 24 hours,cell counting kit-8 was applied to detect the activity of cell proliferation,and real-time PCR was used to detect the expression levels of Asxl1 and osteogenic related genes ALP and RUNX2 mRNA.RESULTS AND CONCLUSION:After lipopolysaccharides stimulation of MC3T3-E1 cells,the expression levels of Asxl1 protein and mRNA were decreased.Under the inflammatory microenvironment,the proliferation activity of MC3T3-E1 cells transfected with Asxl1-Si RNA for 24 hours was significantly decreased,and the expression levels of Asxl1,ALP and RUNX2 mRNA were markedly decreased.These findings indicate that Asxl1 may influence the proliferation and differentiation of osteoblasts by involvement in the process of inflammatory reaction,thereby participating in bone destruction.