论文部分内容阅读
目的:研究牙髓卟啉单胞菌(P.e)和牙龈卟啉单胞菌(P.g)脂多糖(LPS)对成骨细胞表达CD14和TLRs的影响。方法:10μg/mLP.e-LPS和P.g-LPS刺激MC3T3-E1细胞,应用RT-PCR检测不同时间(0、1、3、6、12及24h)后,细胞CD14、TLR2及TLR4 mRNA表达的变化;流式细胞术检测P.e-LPS和P.g-LPS作用24h后,细胞表面CD14、TLR2和TLR4的表达。采用SPSS11.0软件包对结果进行单因素方差分析和Dunnett t检验。结果:P.e-LPS作用1h后,MC3T3-E1细胞CD14和TLR4 mRNA的表达量明显增加,TLR2 mRNA的表达量随着P.e-LPS作用时间的增加并无明显变化。P.g-LPS作用于MC3T3-E1细胞后,其CD14、TLR2、TLR4 mRNA的表达量均明显增加;流式细胞术分析显示,P.e-LPS作用24h后,与未加LPS组相比,CD14和TLR4蛋白的表达量均显著增加,但TLR2蛋白的表达量无显著增加。P.g-LPS作用24h后,细胞CD14、TLR2、TLR4蛋白的表达均显著增加(P<0.05)。结论:P.e-LPS可能是通过CD14和TLR4受体对成骨细胞发挥作用,而P.g-LPS对成骨细胞的刺激作用则可能依赖于CD14、TLR2和TLR4受体。
Objective: To study the effect of P.e and P.g lipopolysaccharide (LPS) on the expression of CD14 and TLRs in osteoblasts. Methods: The MC3T3-E1 cells were stimulated with 10μg / mL P.e-LPS and Pg-LPS. The expressions of CD14, TLR2 and TLR4 mRNA were detected by RT-PCR after different time (0,1,3,6,12 and 24h) Flow cytometry was used to detect the expression of CD14, TLR2 and TLR4 on the cell surface after treatment with Pe-LPS and Pg-LPS for 24 h. One-way ANOVA and Dunnett t test were performed on SPSS 11.0 software package. Results: The expression of CD14 and TLR4 mRNA in MC3T3-E1 cells were significantly increased after P.e-LPS treatment for 1 h, while the expression of TLR2 mRNA did not change significantly with the increase of P.e-LPS effect time. The expression of CD14, TLR2 and TLR4 mRNA in MC3T3-E1 cells were significantly increased after Pg-LPS treatment. Flow cytometry analysis showed that after treated with Pe-LPS for 24 h, the expressions of CD14 and TLR4 Protein expression levels were significantly increased, but no significant increase in TLR2 protein expression. The expression of CD14, TLR2 and TLR4 in P.g-LPS group were significantly increased after 24h (P <0.05). CONCLUSION: P.e-LPS may act on osteoblasts through CD14 and TLR4 receptors, while P.g-LPS may stimulate osteoblasts to depend on CD14, TLR2 and TLR4 receptors.