[髓样分化蛋白-2在脂多糖诱导的人内皮细胞的细胞活化中的作用]

[The role of myeloid differentiation protein-2 in lipopolysaccharide-induced cellular activation of human endothelial cells].

作者信息

Xiong Jian-qiong, Zhu Pei-fang, Wang Zheng-guo, Jiang Jian-xin

机构信息

State Key Laboratory of Trauma, Burns and Combined Injury, Research Institute of Surgery, Daping Hospital, The Third Military Medical University, Chongqing 400042, P.R. China.

出版信息

Zhonghua Shao Shang Za Zhi. 2005 Apr;21(2):97-9.

PMID:15938952

Abstract

OBJECTIVE

To investigate the expression of myeloid differentiation protein-2 (MD-2) in the human endothelial cells and its role in lipopolysaccharide (LPS)-induced NF-kappaB activation in endothelial cells.

METHODS

In vitro cultured human umbilical vein endothelial cells (HUVEC) were employed in the study. The expression of MD-2 mRNA and protein, and the effect of LPS on the expression of its mRNA and protein were assessed with RT-PCR and Western blotting. The role of MD-2 in LPS-induced NF-kappaB activation and IL-8 production were investigated with gene transfection of mutant MD-2 cDNA (0.5, 1.0, 2.0 microg), pEF-BOS vacant vector (2.0 microg) and MD-2 plasmid (2.0 microg) into HUVEC, respectively.

RESULTS

There was MD-2 mRNA and protein expression in HUVECs before LPS stimulation, and it could be obviously upregulated by LPS in time and dose-dependent manner (MD-2 protein absorbency was 25 196 +/- 1 723 without LPS stimulation, which was obviously lower than that stimulated with 0.01 mg/L LPS (58 817 +/- 3 241, P < 0.01) for 6 hours. Transfection of mutant MD-2 cDNA could remarkably inhibit LPS-induced NF-kappaB activation and IL-8 production in endothelial cells.

CONCLUSION

MD-2 might play an important role in the LPS-induced NF-kappaB activation in HUVECs.

摘要

目的

研究髓样分化蛋白-2（MD-2）在人内皮细胞中的表达及其在内皮细胞中脂多糖（LPS）诱导的核因子κB（NF-κB）激活中的作用。

方法

本研究采用体外培养的人脐静脉内皮细胞（HUVEC）。用逆转录聚合酶链反应（RT-PCR）和蛋白质印迹法评估MD-2 mRNA和蛋白的表达以及LPS对其mRNA和蛋白表达的影响。分别将突变型MD-2 cDNA（0.5、1.0、2.0μg）、pEF-BOS空载体（2.0μg）和MD-2质粒（2.0μg）转染入HUVEC，研究MD-2在LPS诱导的NF-κB激活和白细胞介素-8（IL-8）产生中的作用。

结果

LPS刺激前HUVEC中有MD-2 mRNA和蛋白表达，LPS可使其呈时间和剂量依赖性明显上调（无LPS刺激时MD-2蛋白吸光度为25 196±1 723，明显低于0.01 mg/L LPS刺激6小时后的58 817±3 241，P<0.01）。转染突变型MD-2 cDNA可显著抑制LPS诱导的内皮细胞NF-κB激活和IL-8产生。