Elucidation of C-reactive proteins in atherosclerosis

在这个项目中，我们进行了两项研究，以阐明CRP在动脉粥样硬化中的功能作用。在最初的研究中，我们使用了人和兔的标本，试图弄清楚(1)病变中是否存在CRP，(2)如果存在，CRP存放在哪里，(3)血管细胞，特别是巨噬细胞是否可以合成CRP。我们的研究表明，无论病变分期如何，CRP在病变中普遍存在。C反应蛋白与载脂蛋白B密切相关，但与巨噬细胞无关。通过Northern印迹和实时定量RT-PCR，我们证明了CRPmRNA基本上在肝脏中表达，但很少在巨噬细胞表达。我们还发现，在高胆固醇血症兔中，主动脉病变的大小与血浆CRP水平呈正相关。为了证明C反应蛋白是否直接参与病变的形成，我们还在肝脏中产生了表达人C反应蛋白的转基因兔。在这项研究中，我们使用了肝脏特异性表达构建物和绝缘体，并建立了三个独立的CRP转基因兔系。现在，我们正在利用这些转CRP基因的兔来进行动脉粥样硬化的研究。我们还从ISIS公司获得了CRP抑制剂，希望研究抑制CRP是否可以预防动脉粥样硬化。

In this project, we have performed two studies in order to elucidate the functional roles of CRP in atherosclerosis. In the initial study, we used both human and rabbit specimens in attempt to clarify (1) whether CRP is present in the lesions, (2) if so, where CRP is deposited, (3) whether vascular cells especially macrophages can synthesize CRP. Our study showed that CRP is ubiquitously present in the lesions regardless of the lesion stage. CRP is closely associated with apoB but not macrophages. Using both Northern blotting and real-time RT-PCR, we demonstrated that CRP mRNA is basically expressed in the liver but rarely, if any, by macrophages. We also demonstrated that in hypercholesterolemic rabbits, the aortic lesion size is positively correlated with the levels of plasma CRP. In order to demonstrate whether CRP is directly involved in the lesion formation, we also generate transgenic rabbits expressing human CRP in the liver. For this study, we used a liver-specific expression construct along with insulators and established three independent lines of CRP transgenic rabbits. Now, we are using these CRP transgenic rabbits to perform atherosclerosis study. We also obtained CRP inhibitors from ISIS company and hope to investigate whether inhibition of CRP can prevent atherosclerosis.