中华口腔医学杂志
中華口腔醫學雜誌
중화구강의학잡지
Chinese Journal of Stomatology
2008年
3期
146-150
,共5页
龚启梅%凌均棨%蒋宏伟%杜宇%杨芳
龔啟梅%凌均棨%蔣宏偉%杜宇%楊芳
공계매%릉균계%장굉위%두우%양방
受体,CXCR4%细胞增殖%细胞运动%基质细胞衍生因子1
受體,CXCR4%細胞增殖%細胞運動%基質細胞衍生因子1
수체,CXCR4%세포증식%세포운동%기질세포연생인자1
Receptors,CXCR4%Cell proliferation%Cell movement%Stromal cell-derived factor-1
目的 研究体外培养人牙髓细胞(human dental pulp cells,HDPC)上CXCR4的表达情况,检测大肠杆菌脂多糖(lipopolysaccharide,LPS)和肿瘤坏死因子α(TNF-α)刺激后HDPC培养上清液中基质细胞衍生因子1α(stromal cell-derived factor-1α,SDF-1α)的表达水平,探讨人工重组SDF-1α(recombinant human SDF-1α,rhSDF-1α)对HDPC增殖及迁移的影响.方法 采用免疫细胞化学及间接免疫荧光技术检测HDPC上CXCR4的表达.用不同浓度LPS(0.1、1、10、100 mg/L)和TNF-α(1、10、100μg/L)刺激HDPC 48 h后,ELISA法检测HDPC培养上清液中SDF-1α含量的变化.同时甲基噻唑基四唑(MTT)法及体外趋化实验观察不同浓度rhSDF-1α对HDPC增殖及迁移的影响.结果 正常HDPC胞膜表达CXCR4且其培养上清液分泌SDF-1α,浓度约为(4513.55±962.92)ng/L.在用LPS和TNF-α刺激HDPC后,SDF-1α的表达水平均显著降低(P＜0.05).50、100和200μg/L的rhSDF-1α可促进HDPC的增殖(P＜0.05),50和100μg/L rhSDF-1α作用9 h可显著趋化HDPC的迁移(P＜0.01).结论 CXCR4在HDPC上表达且SDF-1α能促进HDPC的增殖及迁移;SDF-1-CXCR4轴可能在牙髓组织损伤修复中发挥重要作用.
目的 研究體外培養人牙髓細胞(human dental pulp cells,HDPC)上CXCR4的錶達情況,檢測大腸桿菌脂多糖(lipopolysaccharide,LPS)和腫瘤壞死因子α(TNF-α)刺激後HDPC培養上清液中基質細胞衍生因子1α(stromal cell-derived factor-1α,SDF-1α)的錶達水平,探討人工重組SDF-1α(recombinant human SDF-1α,rhSDF-1α)對HDPC增殖及遷移的影響.方法 採用免疫細胞化學及間接免疫熒光技術檢測HDPC上CXCR4的錶達.用不同濃度LPS(0.1、1、10、100 mg/L)和TNF-α(1、10、100μg/L)刺激HDPC 48 h後,ELISA法檢測HDPC培養上清液中SDF-1α含量的變化.同時甲基噻唑基四唑(MTT)法及體外趨化實驗觀察不同濃度rhSDF-1α對HDPC增殖及遷移的影響.結果 正常HDPC胞膜錶達CXCR4且其培養上清液分泌SDF-1α,濃度約為(4513.55±962.92)ng/L.在用LPS和TNF-α刺激HDPC後,SDF-1α的錶達水平均顯著降低(P＜0.05).50、100和200μg/L的rhSDF-1α可促進HDPC的增殖(P＜0.05),50和100μg/L rhSDF-1α作用9 h可顯著趨化HDPC的遷移(P＜0.01).結論 CXCR4在HDPC上錶達且SDF-1α能促進HDPC的增殖及遷移;SDF-1-CXCR4軸可能在牙髓組織損傷脩複中髮揮重要作用.
목적 연구체외배양인아수세포(human dental pulp cells,HDPC)상CXCR4적표체정황,검측대장간균지다당(lipopolysaccharide,LPS)화종류배사인자α(TNF-α)자격후HDPC배양상청액중기질세포연생인자1α(stromal cell-derived factor-1α,SDF-1α)적표체수평,탐토인공중조SDF-1α(recombinant human SDF-1α,rhSDF-1α)대HDPC증식급천이적영향.방법 채용면역세포화학급간접면역형광기술검측HDPC상CXCR4적표체.용불동농도LPS(0.1、1、10、100 mg/L)화TNF-α(1、10、100μg/L)자격HDPC 48 h후,ELISA법검측HDPC배양상청액중SDF-1α함량적변화.동시갑기새서기사서(MTT)법급체외추화실험관찰불동농도rhSDF-1α대HDPC증식급천이적영향.결과 정상HDPC포막표체CXCR4차기배양상청액분비SDF-1α,농도약위(4513.55±962.92)ng/L.재용LPS화TNF-α자격HDPC후,SDF-1α적표체수평균현저강저(P＜0.05).50、100화200μg/L적rhSDF-1α가촉진HDPC적증식(P＜0.05),50화100μg/L rhSDF-1α작용9 h가현저추화HDPC적천이(P＜0.01).결론 CXCR4재HDPC상표체차SDF-1α능촉진HDPC적증식급천이;SDF-1-CXCR4축가능재아수조직손상수복중발휘중요작용.
Objective To investigate the expression of CXCR4 in cultured human dental pulp cells (HDPC) in vitro and the corresponding ligand SDF-1α level of HDPC supernatants stimulated by lipopolysaccharide(LPS)and tumor necrosis factor-α(TNF-α),and to explore the role of SDF-1αon the proliferation and the migration of HDPC.Methods The expression of CXCR4 in HDPC was detected by immunocytochemistry technique and indirect immunofluorescence technique.The culture supernatants of HDPC were collected after HDPC had been simulated by LPS and TNF-α of difierent concentrations for 48h and then the SDF-1α level was assayed by quantitative sandwich ELISA.Meanwhile,the effects of recombinant human SDF-1α(rhSDF-1α)on the proliferation and the migration of HDPC at different concentrations were observed by MTT and Boyden Chamber Assay.Results CXCR4 was expressed in cytomembrane of HDPC and SDF-1α was secreted into their normal cell supematants with a concentration of(4513.55±962.92)ng/L. The secretion of SDF-1α was both significantly decreased by stimulation with LPS and TNF-α(P＜0.05).In addition,rhSDF-1α stimulated the HDPC proliferation at the concentrations of 50,100,200μg/L(P＜0.01)and increased the chemotactic migration of HDPC significantly after 9h's incubation with the concentrations of 50,100μg/L(P＜0.05).Conclusions SDF-1α accelerated the proliferation and the migration of HDPC which expressed CXCR4.SDF-1-CXCR4 axis may play a role in repair of pulp injury.
