LPS刺激小鼠骨细胞RANKL/OPG和IL-6表达的实验研究

Effects of LPS on Expression of RANKL/OPG and IL-6 in Mouse Osteocytes in Vitro

目的:探讨LPS对体外培养的小鼠MLO-Y4细胞RANKL/OPG和IL-6表达的影响。方法:以5mg/L的LPS刺激细胞,用CCK-8法于(12、24、48h)后检测细胞的增殖;以不同浓度(1、10、100、500、1000μg/L)的LPS刺激细胞,分别在作用4h和1.5h后用RT-PCR检测细胞对RANKL/OPG和IL-6的相对表达;以100μg/L的LPS刺激细胞,分别在作用(0.5、1、2、4、8h)和(0.5、1、1.5、2、4h)两种不同时间后用RT-PCR检测细胞对RANKL/OPG和IL-6的相对表达。结果:LPS对MLO-Y4细胞增殖无影响;100μg/L的LPS能显著上调细胞对RANKL和IL-6的相对表达,与(500,1000μg/L)LPS的上调结果无统计学差别;除0.5h外其余时间点LPS均上调细胞对RANKL和IL-6的相对表达,并分别在4h和1.5h达到峰值;所有样本LPS对细胞OPG的相对表达均无影响。结论:一定浓度的LPS上调了MLO-Y4细胞对RANKL和IL-6的表达,而对OPG的表达无影响。

Objective：To observe the effects of LPS on the expression of RANKL/OPG and IL-6in MLO-Y4 cells in vitro.Methods：Cells were stimulated with 5 mg/L LPS and the proliferation of the cells was observed at different time points（12,24 and 48h）by CCK-8.Then cells were stimulated with various concentrations of LPS（1,10,100,500 and 1000μg/L）and RT-PCR was performed to detect the relative expression of RANKL/OPG and IL-6after 4hand 1.5h,respectively.Cells were stimulated with 100μg/L LPS and relative expression of RANKL/OPG and IL-6was detected at different time points（0.5,1,2,4and 8h）or（0.5,1,1.5,2and 4h）respectively by RT-PCR.Results：There was no influence of LPS on the proliferation of MLO-Y4 cells.Relative expression of RANKL and IL-6was significantly up-regulated in MLO-Y4 cells stimulated with 100,500and1000μg/L LPS,respectively.When stimulated with 100μg/L LPS,relative expression of RANKL and IL-6was up-regulated at all time points except 0.5h,and RANKL reached peak at 4h,while IL-6at 1.5h.There was no influence of LPS on relative expression of OPG in all samples.Conclusion：Expression of RANKL and IL-6was up-regulated in MLO-Y4 cells in the presence of a certain concentration of LPS,however,OPG was not intervened.