中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2008年
8期
717-721
,共5页
楼小戟%张迪亚%严杰%李盛来%陈莉丽
樓小戟%張迪亞%嚴傑%李盛來%陳莉麗
루소극%장적아%엄걸%리성래%진리려
牙龈卟啉单胞菌%脂多糖%THP-1细胞%炎性细胞因子%Toll样受体
牙齦卟啉單胞菌%脂多糖%THP-1細胞%炎性細胞因子%Toll樣受體
아간계람단포균%지다당%THP-1세포%염성세포인자%Toll양수체
Porphyromonas gingivalis%Lipopolysaccharide%THP-l cell%Inflammation-causing cyto-kines%Toll-like receptor
目的 探讨牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg)脂多糖(LPS)诱导人单核-巨噬样细胞THP-1分泌IL-1β、TNF-α、IL-6能力及相关Toll样受体(TLR)的差异. 方法 采用酚水法提取Pg ATCC33277株脂多糖(Pg-LPS),并用红外光谱法和鲎试验进行鉴定.采用ELISA试剂盒定量检测Pg-LPS作用的THP-1细胞分泌IL-1β、TNF-α和IL-6水平.采用TLR2或TLR4单克隆抗体阻断试验联合ELISA检测,了解Pg-LPS结合靶细胞上Toll样受体的类型.实验中采用大肠杆菌0111:B4株脂多糖(E-LPS)作为对照. 结果 1μg/ml Pg-LPS作用THP-1细胞,分别于0.5、6和6 h检测分泌IL-1β、TNF-α和IL-6的水平明显升高(P<O.01);1 μg/ml E-LPS作用THP-1细胞,分别于6、24和24 h检测分泌IL-1β、TNF-α和IL-6水平亦明显升高(P<0.01).Pg-LPS和E-LPS诱生的上述细胞因子最高浓度相近(P>0.05).TLR2或TLR4单抗均可有效阻断Pg-LPS作用的THP-1细胞分泌IL-1B或IL-6(P<0.05),但仅,ILR2单抗显示了阻断TNF-α分泌的作用(P<0.05).E-LPS诱导THP-1细胞分泌上述3种细胞因子的活性仅可被TLR4单抗所阻断(P<0.05). 结论 Pg-LPS诱导THP-1细胞分泌IL-1β、TNF-α和IL-6活性略高于E-LPS,TLR2而非TLR4是Pg-LPS介导靶细胞分泌上述细胞因子的主要受体.
目的 探討牙齦卟啉單胞菌(Porphyromonas gingivalis,Pg)脂多糖(LPS)誘導人單覈-巨噬樣細胞THP-1分泌IL-1β、TNF-α、IL-6能力及相關Toll樣受體(TLR)的差異. 方法 採用酚水法提取Pg ATCC33277株脂多糖(Pg-LPS),併用紅外光譜法和鱟試驗進行鑒定.採用ELISA試劑盒定量檢測Pg-LPS作用的THP-1細胞分泌IL-1β、TNF-α和IL-6水平.採用TLR2或TLR4單剋隆抗體阻斷試驗聯閤ELISA檢測,瞭解Pg-LPS結閤靶細胞上Toll樣受體的類型.實驗中採用大腸桿菌0111:B4株脂多糖(E-LPS)作為對照. 結果 1μg/ml Pg-LPS作用THP-1細胞,分彆于0.5、6和6 h檢測分泌IL-1β、TNF-α和IL-6的水平明顯升高(P<O.01);1 μg/ml E-LPS作用THP-1細胞,分彆于6、24和24 h檢測分泌IL-1β、TNF-α和IL-6水平亦明顯升高(P<0.01).Pg-LPS和E-LPS誘生的上述細胞因子最高濃度相近(P>0.05).TLR2或TLR4單抗均可有效阻斷Pg-LPS作用的THP-1細胞分泌IL-1B或IL-6(P<0.05),但僅,ILR2單抗顯示瞭阻斷TNF-α分泌的作用(P<0.05).E-LPS誘導THP-1細胞分泌上述3種細胞因子的活性僅可被TLR4單抗所阻斷(P<0.05). 結論 Pg-LPS誘導THP-1細胞分泌IL-1β、TNF-α和IL-6活性略高于E-LPS,TLR2而非TLR4是Pg-LPS介導靶細胞分泌上述細胞因子的主要受體.
목적 탐토아간계람단포균(Porphyromonas gingivalis,Pg)지다당(LPS)유도인단핵-거서양세포THP-1분비IL-1β、TNF-α、IL-6능력급상관Toll양수체(TLR)적차이. 방법 채용분수법제취Pg ATCC33277주지다당(Pg-LPS),병용홍외광보법화후시험진행감정.채용ELISA시제합정량검측Pg-LPS작용적THP-1세포분비IL-1β、TNF-α화IL-6수평.채용TLR2혹TLR4단극륭항체조단시험연합ELISA검측,료해Pg-LPS결합파세포상Toll양수체적류형.실험중채용대장간균0111:B4주지다당(E-LPS)작위대조. 결과 1μg/ml Pg-LPS작용THP-1세포,분별우0.5、6화6 h검측분비IL-1β、TNF-α화IL-6적수평명현승고(P<O.01);1 μg/ml E-LPS작용THP-1세포,분별우6、24화24 h검측분비IL-1β、TNF-α화IL-6수평역명현승고(P<0.01).Pg-LPS화E-LPS유생적상술세포인자최고농도상근(P>0.05).TLR2혹TLR4단항균가유효조단Pg-LPS작용적THP-1세포분비IL-1B혹IL-6(P<0.05),단부,ILR2단항현시료조단TNF-α분비적작용(P<0.05).E-LPS유도THP-1세포분비상술3충세포인자적활성부가피TLR4단항소조단(P<0.05). 결론 Pg-LPS유도THP-1세포분비IL-1β、TNF-α화IL-6활성략고우E-LPS,TLR2이비TLR4시Pg-LPS개도파세포분비상술세포인자적주요수체.
Objective To determine the diversity of Porphytomonas gingivalis lipopolysaccharide (LPS)induced IL-1β,TNF-αand IL-6 levels in THP-1 cells and the associated Toll-like receptors(TLR).Methods P.gingivalis strain ATCC33277 LPS (Pg-LPS)was prepared using phenol-water method and then identified by both infrared spectrometry and limulus test.The levels of IL-1β,TNF-α and IL-6 secreted by THP-1 cells under inducement of Pg-LPS were quantitatively detected using commercial ELISA kits.The bloc-king test using TLR2 or TLR4 monoelonal antibody(McAb)plus the ELISA were used to determine the types of Pg-LPS binding TLR on the surface of target cells.In this study.a commercial LPS from E.coli strain O111:B4(E-LPS)was used as the contr01.Results When 1彬ml ofPg.u,s induced THP.1 ceHs for0.5,6 and 6 h.or l∥rnl ofE-LPs induced for 6,24 and 24 h,respectively,tIIe levels ofTNF.a,IL-1B and IL广6 were in.creased obviously(P<0.01).However,the maximal concentration of tlle t11ree cytokines induced by Pg.LP$ were similar to that induced by E-LPS(P>0.05).tLR2 or TLR4 McAb could block the effects of Pg-LPS in-ducing THP-1 cells to secrete IL-1β and IL-6(P<0.05),where as only TLR2 McAb displayed the inhibition of TNF-α secretion(P<0.05).On the contrast,only TLR4 McAb showed the effects blocking the three cytokines secretion in the THP-1 cells under inducement of E-LPS(P<0.05).Conclusion Pg-LPS shows a slight high-er activity inducing THP-1 cells to secrete IL-1β,TNF-α and IL-6 than E-LPS.TLR2,but not TLR4,is the major receptor of Pg-LPS on the target cells to mediate the secretion of the three cytokines.
