中文摘要：

目的：从基因转录水平抑制小鼠巨噬细胞系Raw264.7细胞表达mfgl2（mouse fibrinogen like protein 2/mfgl2 prothrominase,以下简称mfgl2）和其效应研究.方法：构建mfgl2反义载体（mfgl2-antisense-pcDNA3.0）并转染Raw264.7细胞,在IFN-γ刺激下,分别用RT-PCR、免疫组化、蛋白质印迹分析和mfgl2蛋白功能学检测方法（Procoagulant activity, PCA）检测mfgl2基因转录和蛋白表达水平及PCA的变化.结果：转染mfgl2-antisense-pcDNA3.0可显著抑制Raw264.7细胞mfgl2基因转录和蛋白表达并使其生物学活性显著下降,并且mfgl2表达水平与其PCA活性变化呈平行关系.结论：mfgl2-antisense-pcDNA3.0可有效抑制Raw264.7细胞表达mfgl2,同时其PCA也显著下降,表明Raw264.7细胞PCA与其表达mfgl2有关,此结果为探讨从基因水平治疗与fgl2高表达相关的疾病如重型乙型肝炎、同种移植排斥反应时脏器局部的微血栓形成、纤维素沉积、微循环障碍等病理学损伤寻找新的干预靶点提供了科学依据和手段.

英文摘要：

Objective ：To interfere mfgl2 expression and observe the effects on biological function post mfgl2 gene interfering in mice Raw264. 7 cell line. Methods：A mouse antisense RNA to mfgl2 gene was successfully constructed with a 169 nucleated fragment of mfgl2 cDNA under the CMV promoter of pcDNA3. 1. Mouse macrophage cell line Raw264. 7 were transfected with mfgl2 antisense RNA to show its effect on mfgl2 expression in response to IFN-gema. The biological effects of mfgl2 antisense RNA was measured through RT-PCR, immune histochemistry, Western blot and procogulant activity（ PCA）, the functional assay of mfgl2 protein. Results： A dose dependent inhibitory effect was observed both at the level of mRNA by RT-PCR and protein which was tested by immune histochemistry, western blot as well as PCA. The expression level of mfgl2 corrolated with the changes of PCA in corresponding treated Raw 246. 7 cells. Conclusion：mfgl2 antisense DNA can effectively inhibit the mfgl2 gene expression in Raw264. 7 cells. This study may provide an effective way to interfere the potential therapeutic target mfgl2 gene for diseases with a pathological characteristics of microcirculation disorder including acute on chronic viral hepatitis and acute rejection of allograft, in which mfgl2 may play an important role in its developments of pathology.