中华口腔医学研究杂志（电子版）
中華口腔醫學研究雜誌（電子版）
중화구강의학연구잡지（전자판）
CHINESE JOURNAL OF STOMATOLOGICAL RESEARCH(ELECTRONIC VERSION)
2014年
6期
441-446
,共6页
陈丽红%陈丽虹%蔡艳玲%舒珊%韦曦
陳麗紅%陳麗虹%蔡豔玲%舒珊%韋晞
진려홍%진려홍%채염령%서산%위희
碱性成纤维细胞因子%脂多糖%牙髓细胞%损伤修复
堿性成纖維細胞因子%脂多糖%牙髓細胞%損傷脩複
감성성섬유세포인자%지다당%아수세포%손상수복
Basic fibroblast growth factor%Lipopolysaccharide%Dental pulp cells%Repair of pulp injury
目的：研究不同状态人牙髓组织中碱性成纤维细胞因子(bFGF)的表达特点,以及大肠杆菌脂多糖(LPS)刺激后人牙髓细胞(hDPC)中bFGF的表达水平,探讨bFGF在牙髓损伤修复过程中的可能作用。方法采用实时荧光定量聚合酶链反应(PCR)和免疫蛋白印迹方法(Western blot)分别检测正常、深龋及牙髓炎牙髓组织中bFGF mRNA和蛋白表达情况。实时荧光定量PCR测定1 mg/L LPS刺激hDPC 6、12、24、48 h后bFGF和热休克蛋白70(HSP70)表达水平的变化；Western blot和细胞免疫荧光染色检测LPS刺激hDPC后bFGF蛋白表达变化。结果实时荧光定量PCR和Western blot结果表明,深龋牙髓组织中bFGF水平显著上调,而正常和牙髓炎牙髓组织中bFGF表达无差异。实时荧光定量PCR检测到LPS刺激hDPC后,bFGF和HSP70 mRNA水平同步上调,在12 h 达峰值；Western blot显示,LPS刺激hDPCs 12、24、48 h 后bFGF蛋白表达水平均高于正常hDPC；细胞免疫荧光染色证实,LPS刺激12 h后hDPC中bFGF呈强阳性表达,而正常hDPC中bFGF呈弱阳性表达。结论 bFGF在深龋牙髓组织中高表达,且LPS刺激早期可上调hDPC内bFGF表达,推测bFGF可能参与牙髓组织防御修复反应,可能是细胞抗损伤的重要调节机制之一。
目的：研究不同狀態人牙髓組織中堿性成纖維細胞因子(bFGF)的錶達特點,以及大腸桿菌脂多糖(LPS)刺激後人牙髓細胞(hDPC)中bFGF的錶達水平,探討bFGF在牙髓損傷脩複過程中的可能作用。方法採用實時熒光定量聚閤酶鏈反應(PCR)和免疫蛋白印跡方法(Western blot)分彆檢測正常、深齲及牙髓炎牙髓組織中bFGF mRNA和蛋白錶達情況。實時熒光定量PCR測定1 mg/L LPS刺激hDPC 6、12、24、48 h後bFGF和熱休剋蛋白70(HSP70)錶達水平的變化；Western blot和細胞免疫熒光染色檢測LPS刺激hDPC後bFGF蛋白錶達變化。結果實時熒光定量PCR和Western blot結果錶明,深齲牙髓組織中bFGF水平顯著上調,而正常和牙髓炎牙髓組織中bFGF錶達無差異。實時熒光定量PCR檢測到LPS刺激hDPC後,bFGF和HSP70 mRNA水平同步上調,在12 h 達峰值；Western blot顯示,LPS刺激hDPCs 12、24、48 h 後bFGF蛋白錶達水平均高于正常hDPC；細胞免疫熒光染色證實,LPS刺激12 h後hDPC中bFGF呈彊暘性錶達,而正常hDPC中bFGF呈弱暘性錶達。結論 bFGF在深齲牙髓組織中高錶達,且LPS刺激早期可上調hDPC內bFGF錶達,推測bFGF可能參與牙髓組織防禦脩複反應,可能是細胞抗損傷的重要調節機製之一。
목적：연구불동상태인아수조직중감성성섬유세포인자(bFGF)적표체특점,이급대장간균지다당(LPS)자격후인아수세포(hDPC)중bFGF적표체수평,탐토bFGF재아수손상수복과정중적가능작용。방법채용실시형광정량취합매련반응(PCR)화면역단백인적방법(Western blot)분별검측정상、심우급아수염아수조직중bFGF mRNA화단백표체정황。실시형광정량PCR측정1 mg/L LPS자격hDPC 6、12、24、48 h후bFGF화열휴극단백70(HSP70)표체수평적변화；Western blot화세포면역형광염색검측LPS자격hDPC후bFGF단백표체변화。결과실시형광정량PCR화Western blot결과표명,심우아수조직중bFGF수평현저상조,이정상화아수염아수조직중bFGF표체무차이。실시형광정량PCR검측도LPS자격hDPC후,bFGF화HSP70 mRNA수평동보상조,재12 h 체봉치；Western blot현시,LPS자격hDPCs 12、24、48 h 후bFGF단백표체수평균고우정상hDPC；세포면역형광염색증실,LPS자격12 h후hDPC중bFGF정강양성표체,이정상hDPC중bFGF정약양성표체。결론 bFGF재심우아수조직중고표체,차LPS자격조기가상조hDPC내bFGF표체,추측bFGF가능삼여아수조직방어수복반응,가능시세포항손상적중요조절궤제지일。
Objective To investigate the expression of basic fibroblast growth factor (bFGF) in human dental pulp tissues of different state and the bFGF level in human dental pulp cells (hDPCs) stimulated by lipopolysaccharide (LPS), and explore the potential role of bFGF in the progress of repairing pulp injury. Methods The messenger RNA and protein level of bFGF in normal and inflammatory pulp were detected by fluorescence quantitative-polymerase chain reaction (qPCR) and western blot. HDPCs were collected after stimulated by 1 mg/L LPS for 6, 12, 24 and 48 h. The expression of bFGF and HSP70 in hDPCs was evaluated by qPCR. In addition, the protein level of bFGF in in normal and LPS treated hDPCs were detected by western blot and immunofluorescence staining. Results The mRNA and protein level of bFGF was significantly up-regulated in the pulp with deep caries compared with healthy pulps, while inflamed dental pulp did not show significant difference. QPCR showed that bFGF and HSP70 mRNA were concomitantly increased from 0 h to 12 h after stimulated by LPS. Similarly, it was shown that bFGF was increased in LPS treated hDPCs compared with normal hDPCs by western blot. Moreover, immunofluorescence staining results demonstrated that bFGF was strongly positive stained in LPS treated hDPCs, while it was weakly expressed in normal hDPCs. Conclusion BFGF is up-regulated in pulp with deep caries and hDPCs induced by LPS, indicating that bFGF may participates in the progress of repairing pulp injury.
