中华口腔医学杂志
中華口腔醫學雜誌
중화구강의학잡지
Chinese Journal of Stomatology
2014年
12期
737-741
,共5页
蒋少云%魏丛丛%薛栋%邓嘉胤%连琪%董允允
蔣少雲%魏叢叢%薛棟%鄧嘉胤%連琪%董允允
장소운%위총총%설동%산가윤%련기%동윤윤
紫单胞菌,龈%脂多糖类%牙周炎%肿瘤坏死因子α%白细胞介素1β
紫單胞菌,齦%脂多糖類%牙週炎%腫瘤壞死因子α%白細胞介素1β
자단포균,간%지다당류%아주염%종류배사인자α%백세포개소1β
Porphyromonas gingivalis%Lipopolysaccharides%Periodontitis%Tumor necrosis factor-α%Interleukin-1 β
目的 探讨高浓度葡萄糖对牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg)脂多糖刺激人牙龈成纤维细胞(human gingival fibroblasts,HGF)分泌炎症细胞因子的影响及作用机制,进一步探讨糖尿病与牙周炎的相互关系.方法 组织块法培养HGF,细胞处理分为以下4组:A组:低糖(5.5 mmol/L葡萄糖)+1 mg/L Pg脂多糖刺激组;B组:低糖+10 mg/L Pg脂多糖刺激组;C组:高糖(25 mmol/L葡萄糖)+1 mg/L Pg脂多糖刺激组;D组:高糖+10 mg/L Pg脂多糖刺激组.采用酶联免疫法检测4组HGF细胞6、12h后分泌肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)和白细胞介素1β(interleukin-1β,IL-1β)的变化,实时定量PCR检测Toll样受体(toll-like receptor,TLR)2和TLR4在HGF中的表达.在高糖+10 mg/L Pg脂多糖组采用抗TLR2、抗TLR4单克隆抗体预先处理HGF,检测HGF中12h的TNF-α、IL-1β水平,以高糖+10 mg/L Pg脂多糖刺激组为对照组,并进行统计学分析.结果 相同质量浓度的Pg脂多糖刺激HGF 6、12h后高糖条件(C组、D组)下TNF-α及IL-1β分泌及TLR2 mRNA的表达均较低糖条件下(A组、B组)显著升高(P值均＜0.01),TLR4 mRNA的表达C组与A组相比表达升高,但差异无统计学意义(P＞0.05);D组显著高于B组(P＜0.01).高糖+10 mg/L Pg脂多糖刺激组中分别阻断TLR2、4后,TNF-α的分泌量分别为(297.16± 11.49)、(390.01±12.81) ng/L,均显著低于对照组[(459.80±8.04) ng/L](F=166.02,P＜0.01),IL-1β的分泌量分别为(49.90±4.08)、(99.35±5.01) ng/L,亦均显著低于对照组[(147.37±9.87) ng/L](F=153.51,P＜0.000 1).结论 高糖可明显促进Pg脂多糖刺激HGF后炎症细胞因子的分泌,其作用可能通过调节HGF表面TLR2和TLR4的表达来实现,在一定程度上证实糖尿病患者血糖升高可加重牙周炎症.
目的 探討高濃度葡萄糖對牙齦卟啉單胞菌(Porphyromonas gingivalis,Pg)脂多糖刺激人牙齦成纖維細胞(human gingival fibroblasts,HGF)分泌炎癥細胞因子的影響及作用機製,進一步探討糖尿病與牙週炎的相互關繫.方法 組織塊法培養HGF,細胞處理分為以下4組:A組:低糖(5.5 mmol/L葡萄糖)+1 mg/L Pg脂多糖刺激組;B組:低糖+10 mg/L Pg脂多糖刺激組;C組:高糖(25 mmol/L葡萄糖)+1 mg/L Pg脂多糖刺激組;D組:高糖+10 mg/L Pg脂多糖刺激組.採用酶聯免疫法檢測4組HGF細胞6、12h後分泌腫瘤壞死因子α(tumor necrosis factor-α,TNF-α)和白細胞介素1β(interleukin-1β,IL-1β)的變化,實時定量PCR檢測Toll樣受體(toll-like receptor,TLR)2和TLR4在HGF中的錶達.在高糖+10 mg/L Pg脂多糖組採用抗TLR2、抗TLR4單剋隆抗體預先處理HGF,檢測HGF中12h的TNF-α、IL-1β水平,以高糖+10 mg/L Pg脂多糖刺激組為對照組,併進行統計學分析.結果 相同質量濃度的Pg脂多糖刺激HGF 6、12h後高糖條件(C組、D組)下TNF-α及IL-1β分泌及TLR2 mRNA的錶達均較低糖條件下(A組、B組)顯著升高(P值均＜0.01),TLR4 mRNA的錶達C組與A組相比錶達升高,但差異無統計學意義(P＞0.05);D組顯著高于B組(P＜0.01).高糖+10 mg/L Pg脂多糖刺激組中分彆阻斷TLR2、4後,TNF-α的分泌量分彆為(297.16± 11.49)、(390.01±12.81) ng/L,均顯著低于對照組[(459.80±8.04) ng/L](F=166.02,P＜0.01),IL-1β的分泌量分彆為(49.90±4.08)、(99.35±5.01) ng/L,亦均顯著低于對照組[(147.37±9.87) ng/L](F=153.51,P＜0.000 1).結論 高糖可明顯促進Pg脂多糖刺激HGF後炎癥細胞因子的分泌,其作用可能通過調節HGF錶麵TLR2和TLR4的錶達來實現,在一定程度上證實糖尿病患者血糖升高可加重牙週炎癥.
목적 탐토고농도포도당대아간계람단포균(Porphyromonas gingivalis,Pg)지다당자격인아간성섬유세포(human gingival fibroblasts,HGF)분비염증세포인자적영향급작용궤제,진일보탐토당뇨병여아주염적상호관계.방법 조직괴법배양HGF,세포처리분위이하4조:A조:저당(5.5 mmol/L포도당)+1 mg/L Pg지다당자격조;B조:저당+10 mg/L Pg지다당자격조;C조:고당(25 mmol/L포도당)+1 mg/L Pg지다당자격조;D조:고당+10 mg/L Pg지다당자격조.채용매련면역법검측4조HGF세포6、12h후분비종류배사인자α(tumor necrosis factor-α,TNF-α)화백세포개소1β(interleukin-1β,IL-1β)적변화,실시정량PCR검측Toll양수체(toll-like receptor,TLR)2화TLR4재HGF중적표체.재고당+10 mg/L Pg지다당조채용항TLR2、항TLR4단극륭항체예선처리HGF,검측HGF중12h적TNF-α、IL-1β수평,이고당+10 mg/L Pg지다당자격조위대조조,병진행통계학분석.결과 상동질량농도적Pg지다당자격HGF 6、12h후고당조건(C조、D조)하TNF-α급IL-1β분비급TLR2 mRNA적표체균교저당조건하(A조、B조)현저승고(P치균＜0.01),TLR4 mRNA적표체C조여A조상비표체승고,단차이무통계학의의(P＞0.05);D조현저고우B조(P＜0.01).고당+10 mg/L Pg지다당자격조중분별조단TLR2、4후,TNF-α적분비량분별위(297.16± 11.49)、(390.01±12.81) ng/L,균현저저우대조조[(459.80±8.04) ng/L](F=166.02,P＜0.01),IL-1β적분비량분별위(49.90±4.08)、(99.35±5.01) ng/L,역균현저저우대조조[(147.37±9.87) ng/L](F=153.51,P＜0.000 1).결론 고당가명현촉진Pg지다당자격HGF후염증세포인자적분비,기작용가능통과조절HGF표면TLR2화TLR4적표체래실현,재일정정도상증실당뇨병환자혈당승고가가중아주염증.
Objective To investigate the influence of high glucose on Porphyromonasgingivalis(Pg)lipopolysaccharide(LPS) stimulating human gingival fibroblasts(HGF) to secret the cytokines.Methods HGF were obtained from the primary culture of the tissue explants.Cells were divided into four groups,low glucose(5.5 mmol/L) + 1 mg/L Pg LPS(group A); low glucose+ 10 mg/L Pg LPS(group B); high glucose (25 mmol/L) + 1 mg/L Pg LPS(group C);high glucose+ 10 mg/L Pg LPS(group D).The levels of tumor necrosis factor-α(TNF-α) and interleukin-1β(IL-1β) in cell supernatants were detected by enzyme-linked immunosorbent assay at 6 h and 12 h.The expressions of toll-like receptor 2,4(TLR-2,4) were examined by real-time polymerase chain reaction.After pretreatment with anti-TLR2 and anti-TLR4 monoclonal antibody in HGF,TNF-α and L-1β levels were detected.Results TNF-α concentration increased obviously in high glucose 6 h and 12 h after Pg LPS stimulation(P＜0.01).IL-1β secretion increased(P＜0.01).Meanwhile,TLR2,4 mRNA expression increased,especially in high glucose+ 10 mg/L Pg LPS(P＜0.01).After inhibition of the TLR2,4 in high glucose+ 10 mg/L Pg LPS respectively,TNF-α level[(297.16± 11.49),(390.01 ± 12.81) ng/L] decreased(F=166.02,P＜0.01),and IL-1β level[(49.90±4.08),(99.35±5.01) ng/L] also decreased (F=153.51,P＜0.01).Conclusions High glucose may promote Pg LPS to stimulate the secretion of TNF-α and IL-1β through regulating TLR2,4 expression,which suggests that the elevating blood glucose precipitate in aggravating the process of periodontal disease.
