Altered expression of Bag-1 in Coxsackievirus B3 infected mouse heart
Altered expression of Bag-1 in Coxsackievirus B3 infected mouse heart
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DOI：
10.1016/s0008-6363(00)00323-0
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发表时间：
2001-04-01
影响因子：
10.8
通讯作者：
Archard, LC
中科院分区：
文献类型：
--
作者：
Peng, TQ;Sadusky, T;Archard, LC
目的：科萨基B病毒引起心肌炎或扩张型心肌病的机制尚不清楚。本研究考察了小鼠感染柯萨奇B3病毒（CVB 3）后心脏基因表达的变化。方法：实验性感染小鼠（每组5只），或用稀释液模拟感染。通过cDNA阵列初步鉴定基因表达的改变，并通过半定量RT-PCR、蛋白质印迹和免疫组织化学证实。结果：在含有588个已知小鼠基因的cDNA阵列中，共检测到42个表达上调或下调的基因。其中，一个下调的基因，袋-1，已知参与细胞凋亡的抑制和伴侣活性的调制，进行了进一步的研究。半定量RT-PCR显示，Bag-1表达下调了高达30%，在病毒感染的小鼠心脏在第7天相比，模拟感染。细胞分离和蛋白质印迹分析证实，Bag-1亚型p32主要存在于小鼠心肌细胞质中，并在CVB 3感染后4天或7天下调。相比之下，Bag-1亚型p50在感染后7天小鼠心脏的核部分中似乎增加。Bag-1蛋白的表达和分布下调或细胞凋亡的证据在感染的小鼠心脏中分别通过免疫染色或组织化学（TUNEL法）证实。结论：CVB 3感染可诱导Bag-1在小鼠心肌细胞质和细胞核中的差异表达及细胞凋亡。这可能是肠道病毒心肌病的发病机制中的重要。(C)2001 Elsevier Science B. V.保留所有权利。
Objective: The mechanisms by which Coxsackie B viruses cause myocarditis or dilated cardiomyopathy are not well understood. This study examined changes in the expression of cardiac genes resulting from Coxsackievirus B3 (CVB3) infection of mice. Methods: Mice (five per group) were experimentally infected with CVB3 or mock-infected with diluent. Altered expression of genes was initially identified by cDNA array, and confirmed by semiquantitative RT-PCR, western blot and immunohistochemistry. Results: Forty-two up-regulated or down-regulated genes were observed in cDNA arrays carrying 588 known mouse genes. Among these, one downregulated gene, Bag-1, known to be involved in inhibition of apoptosis and modulation of chaperone activity, was investigated further. Semiquantitative RT-PCR showed that Bag-1 expression was down-regulated by up to 30% in virus-infected mouse heart on day 7 compared to the mock-infected. Cell fractionation and western blot analysis confirmed that Bag-1 isoform p32 was predominant in the cytoplasm of mouse myocardium and down-regulated at 4 days or 7 days after CVB3 infection. In contrast, Bag-1 isoform p50 appeared to increase in the nuclear fraction of mouse heart at 7 days after infection. Down regulated expression and distribution of Bag-1 protein or evidence of apoptosis in the infected mouse heart was demonstrated by immunostaining or histochemistry (TUNEL assay), respectively. Conclusion: CVB3 infection induced differential expression of Bag-1 in cytoplasmic and nuclear fractions of mouse heart and apoptosis. This may be important in the pathogenesis of enterovirus heart muscle disease. (C) 2001 Elsevier Science B.V. All rights reserved.