转化生长因子β3对脂多糖诱导的成骨细胞中IL-6表达的影响  

Inhibiting effect of transforming growth factor β3 on IL-6 expression in MG63 induced by lipopolysaccharide

作　　者：王贵玲[1] 于雅琼[1] 郭佳杰[1] 仇丽鸿[1] WANG Gui-ling YU Ya-qiong GUO Jia-jie QIU Li-hong(Department of Endodontics, School of Stom~alogy, China Medical Universit Liaoning Key Laboratory of Oral Diseases. Shenyang 110002, Liaoning Province, China)

机构地区：[1]中国医科大学口腔医学院牙体牙髓病科,辽宁省口腔疾病重点实验室,辽宁沈阳110002

出　　处：《上海口腔医学》2017年第1期21-25,共5页Shanghai Journal of Stomatology

基　　金：辽宁省自然科学基金(2014021055)

摘　　要：目的 :探讨转化生长因子β3(transforming growth factorβ3,TGF-β3)对成骨细胞内炎性细胞因子IL-6表达的影响,及其发挥抗炎作用的机制。方法:20μg/m L牙髓卟啉单胞菌脂多糖(lipopolysaccharide of Porphyromonas endodontalis,P.e-LPS)刺激人成骨肉瘤细胞系MG63,构建成骨细胞炎症模型。取不同浓度(5~20 ng/m L)的人重组蛋白生长因子TGF-β3和TGF-β1,分别与20μg/m L P.e-LPS共同作用于MG63细胞24 h后,利用实时荧光定量PCR检测细胞内IL-6 m RNA的表达,ELISA法检测培养液上清中IL-6的表达水平。以10 ng/m L TGF-β3预处理细胞30 min后,再与20μg/m L P.e-LPS共同作用20 min,Western印迹法检测细胞内ERK1/2蛋白磷酸化的表达。采用SPSS13.0软件包对数据进行统计学分析。结果:实时荧光定量PCR结果显示,单独20μg/m L P.e-LPS作用下,MG63细胞内IL-6的表达显著增高(P<0.01);TGF-β1在低浓度条件下(5~10 ng/m L)对IL-6的表达无显著作用,仅在20 ng/m L时可显著抑制IL-6的表达(P<0.05)。不同浓度的TGF-β3与P.e-LPS共同作用均可显著抑制IL-6的表达(P<0.01)。ELISA结果显示,10~20 ng/m L TGF-β3可在蛋白水平上对IL-6有显著抑制作用(P<0.05)。单独P.e-LPS作用时,可使MG63细胞内ERK1/2蛋白的磷酸化水平升高(P<0.01);而当TGF-β3与P.e-LPS共同作用时,ERK1/2的磷酸化被抑制(P<0.05)。结论:相同浓度条件下,TGF-β3比TGF-β1对成骨细胞炎症的抑制作用更为显著,ERK1/2信号机制参与了TGF-β3的抗炎过程。PURPOSE： To explore the effect of transforming growth factor β3 （TGF-β3） on IL-6 expression in inflammatory MG63, and the mechanism by which TGF-β3 exert its anti-inflammatory effect. METHODS： Cell line MG63 was stimulated by 20 μg/mL lipopolysaccharide of Porphyromonas endodontalis （P.e-LPS） to establish the inflammatory model of osteoblast. TGF-β3 or TGFβ1 varying from 5 to 20 ng/mL was added together with P.e-LPS for 24 h, then the mRNA expression of IL-6 was detected by real-time PCR, the role of TGF-β3 on IL-6 protein was further verified by ELISA. MG63 was pretreated with 10 ng/mL TGF-β3 for 30 min in RPMI 1640 medium without fetal bovine serum （FBS）, then the cells were cultured for another 20 min with 20 μg/mL P.e-LPS, the phosphorylation level of ERK1/2 was measured by Western blot. Statistical analysis was performed using one-way ANOVA with SPSS13.0 software package. RESULTS： The results of real-time PCR revealed that, when MG63 was treated with 20 μg/mL P.e-LPS alone, the mRNA expression of IL-6 increased significantly（P〈0.01）. When TGF-β1 was added with P.e-LPS, it could barely decrease IL-6 prominently at the highest concentration （P〈0.05）.Whereas, the inhibition effect of TGF-β3 on IL-6 was dramatic （P〈0.01）, ELISA results showed that 10-20 ng/mL TGF-β3 blocked the IL-6 expression at protein level （P〈0.05）. 20 μg/mL P.e-LPS promoted the phosphorylation level of ERK1/2 in MG63（P〈0.01）, while with 10 ng/mL TGF-β3, the effect of P.e-LPS on ERK1/2 was blocked（P〈0.05）. CONCLUSIONS： TGF-β3 is more potent than TGF-β1 in inhibiting MG63, and ERK1/2 is involved in its anti-inflammatory effect.

关 键 词：转化生长因子Β3 成骨细胞 炎症 IL-6 

分 类 号：R781.341[医药卫生—口腔医学]