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[巨噬细胞移动抑制因子在人牙髓中的表达及其对增殖的影响]

[Macrophage migration-inhibitory factors expression and its effects on proliferation in human dental pulps].

作者信息

Zhao Dan-feng, Gong Qi-mei, Ling Jun-qi, Zhang Xu-fang

机构信息

Department of Operative Dentistry and Endodontics, Sun Yat-sen University, Guangzhou, China.

出版信息

Zhonghua Kou Qiang Yi Xue Za Zhi. 2011 Aug;46(8):484-8.

Abstract

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.

摘要

目的

研究巨噬细胞移动抑制因子(MIF)在临床健康和炎症状态下的人牙髓组织中的表达情况,以及重组人巨噬细胞移动抑制因子(rhMIF)对人牙髓细胞(HDPC)增殖的影响。

方法

采用免疫组织化学法检测MIF在临床健康牙髓和炎症牙髓组织中的表达定位。运用定量实时聚合酶链反应(PCR)评估牙髓标本中MIF的mRNA水平。此外,在HDPC经脂多糖(LPS)刺激24小时后收集其培养上清液,然后通过定量夹心酶联免疫吸附测定法检测MIF水平。同时,采用细胞计数试剂盒-8(CCK-8)观察不同浓度的rhMIF作用24小时和48小时对HDPC增殖的影响。

结果

MIF主要分布于健康牙髓组织的成牙本质细胞中,然而,在炎症牙髓组织中,在成纤维细胞、炎症浸润细胞、内皮细胞以及成牙本质细胞中均广泛检测到MIF。定量实时PCR显示,炎症牙髓与健康牙髓的MIF mRNA水平无显著差异(P>0.05)。此外,0.1和1.0mg/L的LPS刺激显著增加了MIF的分泌[(1772.58±495.05),(1692.58±337.45)ng/L](P<0.05),对照组的浓度为(1048.53±161.81)ng/L。rhMIF在10、30、60μg/L浓度下作用24小时和48小时可刺激HDPC增殖。

结论

MIF在牙髓组织中表达,LPS刺激后其表达增加。rhMIF增加了HDPC的增殖。这些结果表明MIF可能参与牙髓炎症过程。

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