Local Gene Silencing of Monocyte Chemoattractant Protein-1 Prevents Vulnerable Plaque Disruption in Apolipoprotein E-Knockout Mice
Local Gene Silencing of Monocyte Chemoattractant Protein-1 Prevents Vulnerable Plaque Disruption in Apolipoprotein E-Knockout Mice
复制标题
单核细胞趋化蛋白-1 的局部基因沉默可防止载脂蛋白 E 敲除小鼠中的易损斑块破坏
DOI：
10.1371/journal.pone.0033497
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发表时间：
2012-03-12
期刊：
影响因子：
3.7
通讯作者：
Zhang, Yun
中科院分区：
文献类型：
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作者：
Liu, Xiao Ling;Zhang, Peng Fei;Zhang, Yun
单核细胞趋化蛋白-1（MCP-1）是一种CC趋化因子（CCL 2），在动脉粥样硬化中发挥重要作用，已成为动脉粥样硬化的重要治疗靶点。本研究旨在验证以下假设：通过腺病毒介导的小发夹RNA（shRNA）的位点特异性递送，MCP-1的局部RNAi可以增强ApoE−/−小鼠中的斑块稳定性并防止斑块破坏。我们设计了一种腺病毒介导的针对小鼠MCP-1的shRNA（rAd 5-MCP-1-shRNA）。雄性载脂蛋白E基因敲除（ApoE−/−）小鼠（n = 120）喂食高脂饮食，通过在颈动脉周围血管周围放置收缩环、腹腔内注射脂多糖和应激刺激诱导易损斑块。 将小鼠随机分为RNA干扰组（Ad-MCP-1 i）、重组腺病毒组（Ad-EGFP）和空白对照组（mock组），分别给予rAd 5-MCP-1-shRNA悬液、rAd 5介导的阴性shRNA和生理盐水。治疗后2周，Ad-MCP-1 i组斑块破裂率明显低于Ad-EGFP组（13.3%vs.60.0%，P = 0.01），免疫组化、qRT-PCR和western blot证实Ad-MCP-1 i组局部MCP-1表达明显受到抑制（P<0.001）。 与Ad-EGFP组相比，Ad-MCP-1 i组颈动脉斑块中胶原和平滑肌细胞水平升高，脂质和巨噬细胞水平降低。Ad-MCP-1 i组炎性细胞因子的表达和基质金属蛋白酶（MMPs）活性均低于Ad-EGFP组。总之，靶向小鼠MCP-1的腺病毒介导的shRNA的位点特异性递送下调MCP-1表达，将易损斑块转变为更稳定的斑块表型并防止斑块破裂。局部炎性细胞因子表达的显著抑制可能是涉及的中心机制。
Monocyte chemoattractant protein-1 (MCP-1), a CC chemokine (CCL2), has been demonstrated to play important roles in atherosclerosis and becoming an important therapeutic target for atherosclerosis. The present study was undertaken to test the hypothesis that local RNAi of MCP-1 by site-specific delivery of adenovirus-mediated small hairpin RNA (shRNA) may enhance plaque stability and prevent plaque disruption in ApoE−/− mice. We designed an adenovirus-mediated shRNA against mouse MCP-1 (rAd5-MCP-1-shRNA). Male apolipoprotein E-knockout (ApoE−/−) mice (n = 120) were fed a high-fat diet and vulnerable plaques were induced by perivascular placement of constrictive collars around the carotid artery, intraperitoneal injection of lipopolysaccharide and stress stimulation. Mice were randomly divided into RNA interference (Ad-MCP-1i) group receiving local treatment of rAd5-MCP-1-shRNA suspension, Ad-EGFP group receiving treatment of rAd5-mediated negative shRNA and mock group receiving treatment of saline. Two weeks after treatment, plaque disruption rates were significantly lower in the Ad-MCP-1i group than in the Ad-EGFP group (13.3% vs. 60.0%, P = 0.01), and local MCP-1 expression was significantly inhibited in the Ad-MCP-1i group confirmed by immunostaining, qRT-PCR and western blot (P<0.001). Compared with the Ad-EGFP group, carotid plaques in the Ad-MCP-1i group showed increased levels of collagen and smooth muscle cells, and decreased levels of lipid and macrophages. The expression of inflammatory cytokines and activities of matrix metalloproteinases (MMPs) were lower in the Ad-MCP-1i group than in the Ad-EGFP group. In conclusion, site-specific delivery of adenoviral-mediated shRNA targeting mouse MCP-1 downregulated MCP-1 expression, turned a vulnerable plaque into a more stable plaque phenotype and prevented plaque disruption. A marked suppression of the local inflammatory cytokine expression may be the central mechanism involved.