【作者】 仉红； 王丽娜； 左美娜； 董明； 史东梅； 徐慧君； 牛卫东；

【Author】 Zhang Hong;Wang Lina;Zuo Meina;Dong Ming;Shi Dongmei;Xu Huijun;Niu Weidong;Dalian Medical University;

【机构】 大连医科大学口腔医学院牙体牙髓科；

【摘要】 目的:观察布鲁顿酪氨酸激酶(BTK)对破骨细胞的增殖及分化的影响,探讨BTK对根尖周炎骨破坏的作用。方法:将RAW264.7用RANKL诱导5天后,通过光学显微镜下观察、TRAP染色及RT-qPCR的方法,验证破骨细胞是否诱导成功;通过转染BTK-Si RNA到诱导5天的细胞内24小时后,利用RT-qPCR检测TRAP mRNA的表达,CCK-8和抗酒石酸酸性磷酸酶(TRAP)检测试剂盒(PNP微板法)检测破骨细胞的增殖及分化情况,并进行统计学分析。结果:RAW264.7用RANKL诱导5天后,光镜下可见大量体积较大,外观呈圆形、梭形、扇形、不规则突起状及TRAP染色呈阳性的多核破骨细胞,经BTK-Si RNA转染24小时后TRAP mRNA的表达水平显著降低(P<0.05);CCK-8和TRAP酶活性检测结果显示破骨细胞增殖及分化能力降低(P<0.05)。结论:抑制BTK后可以抑制破骨细胞的增殖及分化,BTK可作为抑制破骨细胞的新靶点。更多还原

【Abstract】 Objective:to observe the effect of BTK on the proliferation and differentiation of osteoclasts and to explore the mechanism of BTK on bone destruction in periapical periodontitis.Methods:After induction of RAW264.7 with RANKL for 5 days,osteoclast induction was confirmed by light microscopy,TRAP staining,and RT-qPCR;after transfection of BTK-Si RNA into cells induced for 5 days,24 hours later Using RT-qPCR to detect the expression of TRAP mRNA,CCK-8 and tartrate-resistant acid phosphatase(TRAP) detection kit(PNP microplate method)to detect the proliferation and differentiation of osteoclasts,and perform statistical analysis.Results:After RAW264.7 was induced with RANKL for 5 days,a large number of macrophages,which were round,fusiform,fan-shaped,irregularly protuberant,and TRAP-positive,were observed under light microscopy.BTK-Si RNA The expression of TRAP mRNA was significantly reduced after 24 hours of transfection(P<0.05).The detection of CCK-8 and TRAP activity showed that osteoclast proliferation and differentiation were decreased(P<0.05).Conclusion:Inhibition of BTK can inhibit the proliferation and differentiation of osteoclasts.BTK can be used as a new target for the inhibition of osteoclasts.更多还原