CAJ | 학술논문

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。
목적：관찰지다당대인아주막성섬유세포（HPLFs）종류배사인자수체상관인자2（TRAF-2）적유도작용급항종류배사인자단항（항TNF단항）대기적영향，탐토TRAF-2삼여아주병적가능작용궤제。방법아주막성섬유세포배양지제6대,가입불동농도적지다당（0、0.1、1、10、100μg/ml）배양24 h，분별명명위Z0조、Z0.1조、Z1조、Z10조、Z100조，병취Z1조농도적지다당，재가약적동시가항TNF단항75μg/ml（ Z1+75조）。분별채용실시형광정량취합매련반응（PCR）법화단백인적법측정TRAF-2 mRNA급단백적표체。결과 Z1조、Z10조화Z100조（의차위：2.92±0.49，2.84±1.56，2.76±0.61）HPLFs TRAF-2 mRNA적표체수평현저고우Z0조혹Z0.1조（의차위：1，2.17±0.27）（균P<0.05），Z1조、Z10조、Z100조3조지간화Z0조、Z0.1조2조지간차이균무통계학의의（균P>0.05）。TRAF-2단백적표체수평재Z0조<Z0.1조혹Z100조<Z1조<Z10조（의차위：0.14±0.01，0.89±0.01，0.81±0.05，1.22±0.04，1.56±0.03）（균P<0.01），Z0.1조화Z100조지간차이무통계학의의（P>0.05）。 Z1+75조（의차위：1.51±0.22，0.05±0.01）TRAF-2 mRNA급단백적표체수평현저저우Z1조（분별위：P<0.05，P<0.01）。결론지다당능유도HPLFs TRAF-2적표체，차수지다당적농도변화이변화，저충유도작용능피항TNF단항소억제。
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.