【作者】 刘大伟；

【导师】 尚小明；

【作者基本信息】 河北医科大学 ， 内科学， 2011， 硕士

【摘要】 背景:冠心病(Coronary artery disease, CAD)是指冠状动脉粥样硬化使血管腔狭窄或阻塞,或(和)因冠状动脉功能性改变(痉挛)导致心肌缺血缺氧或坏死而引起的心脏病。动脉壁血管平滑肌细胞(Vascular smooth muscle cells,VSMC)是冠状动脉粥样硬化斑块中的主要细胞成分,VSMC的增殖和迁移是引起经皮冠状动脉介入(Percutaneous coronary intervention,PCI)术后再狭窄的关键因素之一,也是CAD的一个主要病因。近年来研究表明,VSMC表型转化是VSMC增殖和迁移的关键性起始步骤,是该类疾病的共同的发病基础。他汀类药物,即3-羟基-3-甲基戊二酰辅酶A还原酶抑制剂,具有降脂、抗炎、抗动脉粥样硬化、抗增殖、抗氧化应激、抗血栓形成、抑制新生血管形成、升高血浆脂联素水平等多重作用,能够保护受损血管内皮。而他汀类药物能否通过抑制血管平滑肌细胞表型转化从而发挥抗动脉粥样硬化的作用,国内外报道尚不多见。目的:本研究拟通过体外培养VSMC,予以血小板源性生长因子(platelet derived growth factor,PDGF)刺激使其由收缩型向合成型转化,使用以瑞舒伐他汀干预,观察瑞舒伐他汀能否抑制VSMC表型转化进而发挥抗动脉粥样硬化的作用。方法:(1)原代培养VSMC及鉴定(2)10nmol/mL PDGF刺激血管平滑肌细胞(3)不同浓度的瑞舒伐他汀对PDGF刺激后的血管平滑肌细胞进行干预(4)免疫细胞化学方法测定血管平滑肌细胞内α-SMA和OPN的表达结果:(1)与对照组相比,经10nmol/mL PDGF诱导脐静脉VSMC48小时后,α-SMA表达下调,差异具有统计学意义(p<0.05);与PDGF诱导组相比,1μmol/L瑞舒伐他汀干预组α-SMA表达上调,差异具有统计学意义(p <0.05);与PDGF诱导组相比,10μmol/L瑞舒伐他汀干预组α-SMA表达上调,差异具有统计学意义(p<0.05)。(2)与对照组相比,经10nmol/mL PDGF诱导静脉VSMC48小时后,OPN表达上调,差异具有统计学意义(p<0.05);与PDGF诱导组相比,1μmol/L瑞舒伐他汀干预组α-SMA表达下调,差异具有统计学意义(p <0.05);与PDGF诱导组相比,10μmol/L瑞舒伐他汀干预组α-SMA表达下调,差异具有统计学意义(p<0.05)。结论:(1)PDGF能够诱导VSMC由收缩型转变为合成型,表现为α-SMA表达下调,OPN表达上调。(2)瑞舒伐他汀能够抑制PDGF诱导的人脐静脉VSMC表型转化,表现为α-SMA表达的上调,OPN表达的下调。(3)本研究表明,高剂量与低剂量瑞舒伐他汀均能抑制VSMC表型转化,高剂量优于低剂量。更多还原

【Abstract】 Background: Coronary artery disease is refers to coronary atherosc- lerotic ascular antrum stricture or obstruction, and coronary artery functional change cause myocardial ischemia anoxia or necrosis and cause heart attacks. The vascular smooth muscle cells(VSMC) are the main cell components of coronary atherosclerotic plaque, phenotype transformation of vascular smooth muscle cells is the key beginning step of the proliferaion and migration of vascular smooth muscle cells in coronary atherosclerotic heart disease with or without stent restenosis (ISR). The statins, 3-Hydroxy-3-Methylg-lutaryl-coenzyme A (HMG CoA) reductase inhibitors, are the most classical and effective cholesterol-lowering drugs. Many studies show that the statins have series of effects on anti-inflammation, anti-atherosclerosis, anti-proliferative, anti-oxidant stress, antithrombotic, the inhibition of angiogenesis and elevated level of plasma adiponectin to protect endothelial from damage. However, it is not yet reported whether the Statins can inhibit the phenotype transformation of vascular smooth muscle to prevent atherosclerosis.Objective: Our studies aim to observe the effect of rosuvastatin on preventing atherosclerosis by inhibiting the phenotype transformation of human umbilical arterial smooth muscle cells in vitro.Methods: 1.The original generation culture VSMC and identifycati- on. 2.10nmol/mL PDGF induction VSMC. 3. 1μmmol/l and 10μmmol/l rosuvastatin intervention VSMC. 4. The expression ofα-actin and osteop- ontin were performed by immunohistochemical Method.Results: 1 The expression ofα-SMA in is significantly decreased in PDGF induction group compared with control group(P<0.05). Rosuvastatin promote the up-regulation ofα-SMA relative to PDGF induction group. The expression levels ofα-SMA in 1and 10μmmol/l Rosuvastatin intervention groups are higher of PDGF induction group respectively, but both are lower than that in control group. There are significantly difference among them (p <0.05). 2 The expression of OPN in is significantly decreased in PDGF induction group compared with control group(P<0.05).Rosuvastatin promote the down-regulation of OPN relative to PDGF induction group. The expression levels of OPN in 1and 10μmmol/l Rosuvastatin intervention groups are lower of PDGF induction group respectively, but both are higher than that in control group. There are significantly difference among them (p <0.05).Conclusions: 1 PDGF promotes the transformation of VSMC pheno -type from contractile to synthesize phenotype. 2 The expression ofα-ac tin was significantly decreased, while the expression of osteop-ontin was significantly increased during the process of phenotype transformation. 3 high dose and low dose Rosuvastatin may inhibit the transformation of VSMC phenotype from contractile to synthesize phenotype with the association of elevated level ofα-actin and decreased level of osteopontin. And high dose is much more effective更多还原