中华口腔医学杂志
中華口腔醫學雜誌
중화구강의학잡지
Chinese Journal of Stomatology
2013年
12期
730-733
,共4页
雷浪%李厚轩%潘盛波%闫福华
雷浪%李厚軒%潘盛波%閆福華
뢰랑%리후헌%반성파%염복화
紫单胞菌,龈%巨噬细胞%髓系细胞触发受体1
紫單胞菌,齦%巨噬細胞%髓繫細胞觸髮受體1
자단포균,간%거서세포%수계세포촉발수체1
Porphyromonas gingivalis%Macrophages%Triggering receptors expressed on myeloid-1
目的 研究髓系触发细胞受体1(triggering receptors expressed on myeloid-1,TREM-1)是否参与巨噬细胞对牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg)的先天免疫反应,以进一步分析TREM-1在牙周病发病中的作用.方法 分离培养小鼠腹腔巨噬细胞,空白对照组以含10％胎牛血清的RPMI 1640培养液培养,实验组加入Pg刺激;实时荧光定量PCR分析TREM-1基因转录水平的变化,流式细胞术检测其蛋白水平的变化;采用LP-17多肽(10、100和1000 μg/L)特异性阻断TREM-1的作用,酶联免疫吸附测定法检测培养液中肿瘤坏死因子α(tumor necrosis factor-α,TNF-αα)和白细胞介素6(interleukin-6,IL-6)水平的变化.结果 细菌刺激巨噬细胞2h后,TREM-1基因转录水平上调(7.99±1.11)倍;细菌刺激24 h后,流式细胞检测显示TREM-1蛋白表达水平上调:平均荧光强度空白对照组为(7.05±1.85),实验组为(13.17±2.33);100和1000 μg/L的LP-17多肽阻断TREM-1后,TNF-α和IL-6的分泌下降.结论 TREM-1促进了巨噬细胞对Pg的先天免疫反应,参与了牙周病的发病过程.
目的 研究髓繫觸髮細胞受體1(triggering receptors expressed on myeloid-1,TREM-1)是否參與巨噬細胞對牙齦卟啉單胞菌(Porphyromonas gingivalis,Pg)的先天免疫反應,以進一步分析TREM-1在牙週病髮病中的作用.方法 分離培養小鼠腹腔巨噬細胞,空白對照組以含10％胎牛血清的RPMI 1640培養液培養,實驗組加入Pg刺激;實時熒光定量PCR分析TREM-1基因轉錄水平的變化,流式細胞術檢測其蛋白水平的變化;採用LP-17多肽(10、100和1000 μg/L)特異性阻斷TREM-1的作用,酶聯免疫吸附測定法檢測培養液中腫瘤壞死因子α(tumor necrosis factor-α,TNF-αα)和白細胞介素6(interleukin-6,IL-6)水平的變化.結果 細菌刺激巨噬細胞2h後,TREM-1基因轉錄水平上調(7.99±1.11)倍;細菌刺激24 h後,流式細胞檢測顯示TREM-1蛋白錶達水平上調:平均熒光彊度空白對照組為(7.05±1.85),實驗組為(13.17±2.33);100和1000 μg/L的LP-17多肽阻斷TREM-1後,TNF-α和IL-6的分泌下降.結論 TREM-1促進瞭巨噬細胞對Pg的先天免疫反應,參與瞭牙週病的髮病過程.
목적 연구수계촉발세포수체1(triggering receptors expressed on myeloid-1,TREM-1)시부삼여거서세포대아간계람단포균(Porphyromonas gingivalis,Pg)적선천면역반응,이진일보분석TREM-1재아주병발병중적작용.방법 분리배양소서복강거서세포,공백대조조이함10％태우혈청적RPMI 1640배양액배양,실험조가입Pg자격;실시형광정량PCR분석TREM-1기인전록수평적변화,류식세포술검측기단백수평적변화;채용LP-17다태(10、100화1000 μg/L)특이성조단TREM-1적작용,매련면역흡부측정법검측배양액중종류배사인자α(tumor necrosis factor-α,TNF-αα)화백세포개소6(interleukin-6,IL-6)수평적변화.결과 세균자격거서세포2h후,TREM-1기인전록수평상조(7.99±1.11)배;세균자격24 h후,류식세포검측현시TREM-1단백표체수평상조:평균형광강도공백대조조위(7.05±1.85),실험조위(13.17±2.33);100화1000 μg/L적LP-17다태조단TREM-1후,TNF-α화IL-6적분비하강.결론 TREM-1촉진료거서세포대Pg적선천면역반응,삼여료아주병적발병과정.
Objective To investigate the role of triggering receptors expressed on myeloid-1 (TREM-1) in innate response to Porphyromonas gingivalis(Pg) in mice macrophages and its potential role in periodontitis development.Methods Peritoneal macrophages from mice were harvested,separated and cultured,then challenged with viable Pg.Transcription and protein expression in macrophages were assessed with real time PCR and flow cytometry respectively.LP-17 peptide (10,100 and 1000 μg/L) was utilized to block TREM-1,and tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were detected by enzyme linked absorbent analysis.Results At 2 h after Pg challenge,transcription of TREM-1 was significantly upregulated after Pg challenge [(7.99 ± 1.11) fold vs blank].At 24 h after bacteria infection,increased TREM-1 expression was demonstrated by flow cytometry,with mean fluorescent intensity increasing from (7.05 ± 1.85) in blank group to (13.17 ± 2.33) in experimental group.Proinflammatory cytokine(TNF-α and IL-6) production was significantly decreased after blocking TREM-1 by LP-17 peptide (100 and 1000 μg/L).Conclusions TREM-1 enhanced innate immune response to Pg in macrophages,which may facilitate periodontitis development.
