干扰素诱导蛋白p204在大鼠血管壁成分细胞的表达

Expression of the Interferon-inducible Protein p204 in Murine Vessel Wall Cells

目的研究干扰素诱导蛋白p204在鼠血管壁各成分细胞的基础表达及干扰素诱导性。方法通过免疫组织化学染色及Western blot观察p204蛋白在成年昆明小鼠及Wistar大鼠或SD大鼠血管壁的表达;免疫细胞化学法及Western blot观察p204在培养的SD大鼠主动脉血管平滑肌细胞(VSMC)和血管外膜成纤维细胞(VAF)的表达及分布;逆转录-聚合酶链反应观察干扰素诱导VSMC上p204 mRNA(Ifi204)表达特点。结果免疫组织化学染色表明p204抗原在成年昆明小鼠心内小静脉及小动脉血管壁内皮细胞、内膜、中膜及外膜的细胞胞浆中呈阳性反应,p204在成年Wistar大鼠主动脉内皮、VSMC及外膜均染色阳性,弹性纤维膜未着色。Western blot表明p204在SD大鼠主动脉有基础表达,其在VAF的基础表达量明显高于VSMC,免疫细胞化学法表明p204在鼠主动脉VSMC细胞同时表达于细胞质及细胞核。应用干扰素α1.0×106IU/L刺激VSMC细胞8 h即诱导Ifi204大量表达,干扰素α作用24 h时表达更强烈。结论 p204在小鼠及大鼠血管壁全层和培养的VSMC及VAF均存在基础表达,p204在VSMC上表达并不局限于细胞核,且易被干扰素α诱导其mRNA表达上调。p204在鼠血管壁细胞存在表达提示其在血管生理及血管增殖性疾病的发生发展中可能有着潜在的重要作用。

Aim To Study the basic expression of the interferon-inducible protein p200 in murine vessel wall cells, and the characteristic induced by interferon （IFN）. Methods We used immunohistochemistry and Western blot to observe the expression of protein p204 in adult Kunming （KM） mice and Wistar rat or SD rat vessel wall, through immunocytochemistry and Western blot observation studied the expression and distribution of p200 in cultured vascular smooth muscle cells （VSMC） and vascular adventitia fibroblasts （VAF） which was isolated from SD rat aorta, and used re- verse transcription polymerase chain reaction （RT-PCR） to detect the characteristic of IFN-a induced interferon induced protein 200 （Ifi200） mRNA expression. Results Immunohistochemistry showed that p204 antigen stain were positive in vascular endothelial cells （ VEC）, intima, media and adventitia of intracardiac venules and arterioles vessel wall; p204 stain were positive in adult Wistar rat aortic endothelium, VSMC, adventitia, and negative in elastic fiber membrane. Western blot indicated the basic expression of p200 in SD rat aorta, and the VAF p204 basic expression was obviously high- er than VSMC. Immunocytochemistry showed that p204 were present in the cytoplasm and also nucleus of VSMC. Ap-plication of IFN-a with the terminal concentration of 1.0 ×106 IU/L stimulated VSMC cell for 8 hours, that is to say, induced the sig- nificant expression of/fi204 mRNA,then it expressed more intense when stimulated for 24 hours. Conclusions There are basic expressions of p204 in full vessel wall of mice or rat and cultured rat VSMC or VAF. p204 expression is not limited to the nucleus and IFN-a can induce the Ifi204 mRNA expression increased in VSMC. p204 expression in routine vessel wall cells suggests its potential and important role in vascular physiology and also in the occurrence and development of vascular proliferation diseases.