抗肿瘤坏死因子单抗对脂多糖诱导的人牙周膜成纤维细胞TRAF-2的调控作用  

Regulation of anti-tumor necrosis factor antibody on lipopolysaccharide-induced TRAF-2 expression in human periodontal ligament fibroblasts

作　　者：陈焱[1] 许红苗[1] 莫文元 俞诚波[1] 林晡[1] 尹向鹏[1] 

机构地区：[1]温州医学院附属温岭市医院口腔科,317500 [2]浙江省台州市中西医结合医院外科

出　　处：《中国药物与临床》2014年第7期891-894,共4页Chinese Remedies & Clinics

基　　金：浙江省温岭市科技局基金(2013-2-111)

摘　　要：目的观察脂多糖对人牙周膜成纤维细胞(HPLFs)肿瘤坏死因子受体相关因子2(TRAF-2)的诱导作用及抗肿瘤坏死因子单抗(抗TNF单抗)对其的影响,探讨TRAF-2参与牙周病的可能作用机制。方法牙周膜成纤维细胞培养至第6代,加入不同浓度的脂多糖(0、0.1、1、10、100μg/ml)培养24 h,分别命名为Z0组、Z0.1组、Z1组、Z10组、Z100组,并取Z1组浓度的脂多糖,在加药的同时加抗TNF单抗75μg/ml(Z1+75组)。分别采用实时荧光定量聚合酶链反应(PCR)法和蛋白印迹法测定TRAF-2 mRNA及蛋白的表达。结果 Z1组、Z10组和Z100组(依次为:2.92±0.49,2.84±1.56,2.76±0.61)HPLFs TRAF-2 mRNA的表达水平显著高于Z0组或Z0.1组(依次为:1,2.17±0.27)(均P<0.05),Z1组、Z10组、Z100组3组之间和Z0组、Z0.1组2组之间差异均无统计学意义(均P>0.05)。TRAF-2蛋白的表达水平在Z0组<Z0.1组或Z100组<Z1组<Z10组(依次为:0.14±0.01,0.89±0.01,0.81±0.05,1.22±0.04,1.56±0.03)(均P<0.01),Z0.1组和Z100组之间差异无统计学意义(P>0.05)。Z1+75组(依次为:1.51±0.22,0.05±0.01)TRAF-2 mRNA及蛋白的表达水平显著低于Z1组(分别为:P<0.05,P<0.01)。结论脂多糖能诱导HPLFs TRAF-2的表达,且随脂多糖的浓度变化而变化,这种诱导作用能被抗TNF单抗所抑制。Objective To investigate the effects of lipopolysaccharide on secretion of tumor necrosis factor re-ceptor-associated factor-2 （TRAF2） and the regulation by anti-tumor necrosis factor （TNF） antibody on lipopolysaccha-ride-induced TRAF2 expression of human periodontal ligament fibroblasts （HPLFs）, thus exploring the putative mech-anisms of TRAF-2 in the pathogenesis of periodontal diseases. Methods HPLFs were cultured to the sixth genera-tion and incubated with various concentrations of lipopolysaccharide （0, 0.1, 1, 10 and 100 ug/ml） for 24h （groups Z0, Z0.1, Z1, Z10 and Z100）. The Z1 group was incubated with 75 μg/ml anti-TNF antibody （group Z1＋75）. The expres-sions of TRAF-2 mRNA and protein were analyzed by real-time quantitative reverse transcription polymerase chain reaction and Western blotting assay. Results The expressions of TRAF-2 mRNA in groups Z1, Z10 and Z100 （2.92± 0.49, 2.84±1.56 and 2.76±0.61） were significantly higher than those in groups Z0 and Z0.1 （1.00 and 2.17 ±0.27, all P〈0.05）. However, the difference in TRAF-2 mRNA expression among groups Z1, Z10 and Z100 and between group Z0 and Z0.1 was unremarkable （all P〉0.05）. The expression level of TRAF-2 protein was 0.14 ±0.01 in group Z0, 0.89±0.01 in group Z0.1, 0.81±0.05 in group Z100, 1.22±0.04 in group Z1, and 1.56±0.03 in group Z10, respectively （all P〈0.01）. The between-group differences were significant except for between group Z0.1 and Z100 （ P〉0.05）. Fur-thermore, expression levels of TRAF-2 mRNA and protein in group Z1＋75 （1.51±0.22 and 0.05±0.01） were signifi-cantly lower than those in group Z1 （P〈0.05 and P〈0.01）. Conclusion TRAF-2 expressions in HPLFs can be in-duced by lipopolysaccharide. Chnages in TRAF-2 expression with different concentrations of lipopolysaccharide can be abrogated by anti-TNF antibody.

关 键 词：牙周膜 成纤维细胞 脂多糖类 牙周病学 TRAF-2 

分 类 号：R781.4[医药卫生—口腔医学]