中华口腔医学杂志
中華口腔醫學雜誌
중화구강의학잡지
Chinese Journal of Stomatology
2011年
8期
484-488
,共5页
赵丹凤%龚启梅%凌均棨%张旭芳
趙丹鳳%龔啟梅%凌均棨%張旭芳
조단봉%공계매%릉균계%장욱방
牙髓炎%巨噬细胞游走抑制因子%牙髓细胞
牙髓炎%巨噬細胞遊走抑製因子%牙髓細胞
아수염%거서세포유주억제인자%아수세포
Pulpitis%Macrophage migration-inhibitory factors%Dental pulp cells
目的 研究正常及炎症牙髓组织中巨噬细胞游走抑制因子(macrophage migration-inhibitory factors,MIF)的表达及其对人牙髓细胞(human dental pulp cells,HDPC)增殖的影响,以期探讨MIF在牙髓炎症中的可能作用.方法采用免疫组织化学和实时荧光定量聚合酶链反应(PCR)法检测健康及炎症牙髓组织中MIF的表达情况,并以不同质量浓度[0(对照组)、0.1、1.0、10.0 mg/L]脂多糖刺激HDPC 24 h,ELISA法检测HDPC培养上清液中MIF含量的变化;不同质量浓度(10、30、60 μg/L)的重组人MIF分别作用于HDPC 24和48 h,细胞计数试剂盒法检测细胞增殖率.结果 健康牙髓组织中MIF主要分布于成牙本质细胞层,炎症牙髓组织中MIF分布于成牙本质细胞、炎症细胞、牙髓成纤维细胞和血管内皮细胞中;MIF mRNA在正常牙髓组织和炎症牙髓组织中的表达差异无统计学意义(P>0.05);不同质量浓度脂多糖刺激HDPC后对照组MIF质量浓度为(1048.53±161.81) ng/L,0.1和1.0 mg/L组MIF分泌量[分别为(1772.58±495.05)、(1692.58±337.45) ng/L]与对照组相比均显著升高(P<0.05),约为对照组的1.5倍;10、30、60 μg /L 重组人MIF均可促进HDPC的增殖(P<0.05).结论 MIF在人牙髓组织中有表达且一定浓度脂多糖可促进HDPC MIF的分泌,重组人MIF可促进HDPC增殖,MIF可能在牙髓炎症发展过程中发挥一定作用.
目的 研究正常及炎癥牙髓組織中巨噬細胞遊走抑製因子(macrophage migration-inhibitory factors,MIF)的錶達及其對人牙髓細胞(human dental pulp cells,HDPC)增殖的影響,以期探討MIF在牙髓炎癥中的可能作用.方法採用免疫組織化學和實時熒光定量聚閤酶鏈反應(PCR)法檢測健康及炎癥牙髓組織中MIF的錶達情況,併以不同質量濃度[0(對照組)、0.1、1.0、10.0 mg/L]脂多糖刺激HDPC 24 h,ELISA法檢測HDPC培養上清液中MIF含量的變化;不同質量濃度(10、30、60 μg/L)的重組人MIF分彆作用于HDPC 24和48 h,細胞計數試劑盒法檢測細胞增殖率.結果 健康牙髓組織中MIF主要分佈于成牙本質細胞層,炎癥牙髓組織中MIF分佈于成牙本質細胞、炎癥細胞、牙髓成纖維細胞和血管內皮細胞中;MIF mRNA在正常牙髓組織和炎癥牙髓組織中的錶達差異無統計學意義(P>0.05);不同質量濃度脂多糖刺激HDPC後對照組MIF質量濃度為(1048.53±161.81) ng/L,0.1和1.0 mg/L組MIF分泌量[分彆為(1772.58±495.05)、(1692.58±337.45) ng/L]與對照組相比均顯著升高(P<0.05),約為對照組的1.5倍;10、30、60 μg /L 重組人MIF均可促進HDPC的增殖(P<0.05).結論 MIF在人牙髓組織中有錶達且一定濃度脂多糖可促進HDPC MIF的分泌,重組人MIF可促進HDPC增殖,MIF可能在牙髓炎癥髮展過程中髮揮一定作用.
목적 연구정상급염증아수조직중거서세포유주억제인자(macrophage migration-inhibitory factors,MIF)적표체급기대인아수세포(human dental pulp cells,HDPC)증식적영향,이기탐토MIF재아수염증중적가능작용.방법채용면역조직화학화실시형광정량취합매련반응(PCR)법검측건강급염증아수조직중MIF적표체정황,병이불동질량농도[0(대조조)、0.1、1.0、10.0 mg/L]지다당자격HDPC 24 h,ELISA법검측HDPC배양상청액중MIF함량적변화;불동질량농도(10、30、60 μg/L)적중조인MIF분별작용우HDPC 24화48 h,세포계수시제합법검측세포증식솔.결과 건강아수조직중MIF주요분포우성아본질세포층,염증아수조직중MIF분포우성아본질세포、염증세포、아수성섬유세포화혈관내피세포중;MIF mRNA재정상아수조직화염증아수조직중적표체차이무통계학의의(P>0.05);불동질량농도지다당자격HDPC후대조조MIF질량농도위(1048.53±161.81) ng/L,0.1화1.0 mg/L조MIF분비량[분별위(1772.58±495.05)、(1692.58±337.45) ng/L]여대조조상비균현저승고(P<0.05),약위대조조적1.5배;10、30、60 μg /L 중조인MIF균가촉진HDPC적증식(P<0.05).결론 MIF재인아수조직중유표체차일정농도지다당가촉진HDPC MIF적분비,중조인MIF가촉진HDPC증식,MIF가능재아수염증발전과정중발휘일정작용.
Objective To investigate the expression of macrophage migration-inhibitory factors(MIF) in clinically healthy and inflamed human pulp tissues and the effects of rhMIF on the proliferation of human dental pulp cells(HDPC). Methods Immunohistochemistry was used to detect the localization of MIF expression in clinically healthy pulp and inflamed pulp tissues. Quantitative real-time polymerase chain reaction(PCR) was performed to evaluate the mRNA levels of MIF in pulp specimens. In addition, the culture supernatants of HDPC were collected after HDPC was stimulated by lipopolysaccharide(LPS) for 24 h, and then the MIF levels were assayed by quantitative sandwich enzyme-linked immunosorbent assay. Meanwhile, the effects of rhMIF on the proliferation of HDPC at different concentrations for 24 and 48 h were observed by cell counting kit-8(CCK-8). Results MIF was mainly distributed in odontoblasts of healthy pulp tissue, however, in inflamed pulp tissue, it was widely detected in fibroblasts, inflammatory infiltrates and endothelial cells as well as odontoblasts. Quantitative real-time PCR showed that there was no significant difference in MIF mRNA levels between inflamed pulps and healthy pulps(P>0.05). Additionally, the secretion of MIF was significantly increased by stimulation with LPS at the concentration of 0.1 and 1.0 mg/L[(1772.58±495.05),(1692.58±337.45) ng/L] (P<0.05) ,and the concentration was (1048.53±161.81) ng/L in control group. rhMIF stimulated the HDPC′s proliferation at the concentration of 10,30,60 μg/L for 24 and 48 h. Conclusions MIF was expressed in pulp tissue and its expression was increased after stimulation by LPS. rhMIF increased the proliferation of HDPC. These results suggest that MIF may be involved in the process of pulpal inflammation.
