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骨形态发生蛋白-4在高糖环境下静脉桥增殖重构进程中的作用研究
引用本文:方登峰,张洪伟,古 君,等.骨形态发生蛋白-4在高糖环境下静脉桥增殖重构进程中的作用研究[J].四川大学学报(医学版),2017,48(5):710-715.
作者姓名:方登峰  张洪伟  古 君  
作者单位:1.四川大学华西医院 麻醉科
摘    要:目的 观察糖尿病大鼠自体静脉桥的形态学变化以及骨形态发生蛋白4(bone morphogenic protein4,BMP4)和增殖细胞抗原Ki67在其管壁中的表达与分布情况,初步探讨BMP4在高糖环境下静脉桥增殖重构进程中的潜在作用。方法 选用8周龄的48只雄性SpragueDawley大鼠,随机分配为非糖尿病组(即血糖正常组, n=24)及糖尿病组( n=24),采用高脂高糖饮食联合链脲佐菌素的诱导方法构建糖尿病大鼠模型。两组大鼠均建立单侧颈外静脉颈总动脉桥接模型,每组分别于术前,术后1周、2周和4周获取大鼠静脉桥( n=6)。应用HE染色切片测算大鼠静脉桥血管壁的形态学参数,免疫组化观察Ki67及BMP4在管壁中的表达及分布情况,逆转录聚合酶链式反应(RTPCR)和免疫印迹法(Western blot)检测BMP4基因和蛋白在静脉桥中的表达量。结果 在大鼠静脉桥增殖重构进程中,管壁随时间进展不断增厚,而糖尿病大鼠术后内膜和中膜厚度均高于同期的正常大鼠( P<0.05)。免疫组化提示Ki67表达于大鼠静脉桥管壁平滑肌细胞的胞核,Ki67的阳性表达随桥血管增殖重构进程逐渐增高,而术后糖尿病大鼠静脉桥内Ki67的阳性细胞数高于同期的正常大鼠( P<0.05)。免疫组化提示BMP4表达于大鼠静脉桥管壁平滑肌细胞的胞浆,结合RTPCR及Western blot检测发现,正常大鼠术后静脉桥中仅有少量BMP4的表达,而糖尿病大鼠静脉桥管壁内BMP4的阳性细胞数以及在基因和蛋白水平的表达量均随时间进展逐渐增高,并高于术后同期的正常大鼠( P<0.05)。结论 糖尿病大鼠静脉移植术后桥血管增殖性重构改变与管壁内BMP4的高表达密切相关,而BMP4可能在高糖环境下静脉桥加速失效进程中发挥了重要的促进作用。

关 键 词:骨形态发生蛋白-4  高糖环境  冠状动脉旁路移植术  静脉桥

The Role of Bone Morphogenic Protein-4 in the Progression of Vein Graft Hyperplastic Remodeling under
FANG Deng-feng,ZHANG Hong-wei,GU Jun,et al.The Role of Bone Morphogenic Protein-4 in the Progression of Vein Graft Hyperplastic Remodeling under[J].Journal of West China University of Medical Sciences,2017,48(5):710-715.
Authors:FANG Deng-feng  ZHANG Hong-wei  GU Jun  
Abstract:【Abstract】 Objective This study was designed to assess the vessel wall characteristics and the expression levels of bone morphogenic protein4(BMP4) and proliferating cell antigen Ki67 in vein grafts harvested from diabetic rats, and to investigate the role of BMP4 in progression of vein graft hyperplastic remodeling under hyperglycemic condition. Methods 48 male SpragueDawky rats 〔body mass (194±16) g〕 aged 8 weeks were randomly divided into diabetes mellitus (DM) group ( n=24) and nondiabetes mellitus (NDM) group ( n=24). The DM rats were induced by streptozotocin in combination with highsugar and highfat diet. The unilateral external jugular vein was interposition grafted into the common carotid arteries in the two groups. The vein grafts were harvested at preoperatively and 1, 2 and 4 weeks postoperatively ( n=6) in each group. The morphological characteristics of the venous graft wall were evaluated by hematoxylineosin staining, and the expression levels and the distribution of Ki67 and BMP4 were assessed by immunohistochemistry analysis, then the expression of BMP4 gene and protein was measured by realtime polymerase chain reaction (RTPCR) and Western blot assay respectively. Results In the progression of rats vein grafts hyperplastic remodeling, the venous wall thickness of rats thickened with time increasing, and the intimal and medial thickness of the vein grafts harvested from DM rats were significantly higher than those from NDM rats at the same time postoperatively ( P<0.05). Ki67 was highly xpressed in smooth muscle cells nucleus located in the rats vein grafts, and its expression level was upregulated
Keywords:Bone morphogenic protein-4    Hyperglycemic condition    Coronary artery bypass graft
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