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

发布时间：2018-12-16 03:10

【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 protein-4,BMP4)和增殖细胞抗原Ki-67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Sprague-Dawley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Ki-67及BMP4在管壁中的表达及分布情况,逆转录-聚合酶链式反应(RT-PCR)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(P0.05)。免疫组化提示Ki-67表达于大鼠静脉桥管壁平滑肌细胞的胞核,Ki-67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Ki-67的阳性细胞数高于同期的正常大鼠(P0.05)。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RT-PCR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(P0.05)。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。
[Abstract]:Objective to observe the morphologic changes of autogenous vein bridge and the expression and distribution of bone morphogenetic protein-4 (bone morphogenic protein-4,BMP4) and proliferative cell antigen (Ki-67) in diabetic rats. To explore the potential role of BMP4 in the process of vein bridge proliferation and remodeling in high glucose environment. Methods 48 male Sprague-Dawley rats aged 8 weeks were randomly assigned to non-diabetic group (normal blood glucose group, nong24) and diabetic group (nni24). The diabetic rats were induced by high-fat and high-glucose diet combined with streptozotocin (STZ). The unilateral external jugular vein-common carotid artery bridging model was established in both groups. The venous bridge (nong6) was obtained in each group before operation, 1 week, 2 weeks and 4 weeks after operation, respectively. The morphological parameters of vascular wall of venous bridge in rats were measured by HE staining section. The expression and distribution of Ki-67 and BMP4 in vascular wall were observed by immunohistochemistry. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot (Western blot) were used to detect the expression of BMP4 gene and protein in venous bridge. Results in the process of vein bridge proliferation and remodeling, the wall of the vessel was thickened with time, while the thickness of intima and media of diabetic rats was higher than that of normal rats after operation (P0.05). Immunohistochemical staining showed that Ki-67 was expressed in the nucleus of smooth muscle cells of rat venous bridge wall, and the positive expression of Ki-67 increased with the proliferation and remodeling of grafts. The number of Ki-67 positive cells in vein bridge of diabetic rats was higher than that of normal rats (P0.05). Immunohistochemical staining showed that BMP4 was expressed in the cytoplasm of smooth muscle cells of rat venous bridge wall. Combined with RT-PCR and Western blot detection, only a small amount of BMP4 expression was found in normal rat vein bridge after operation. However, the number of BMP4 positive cells and the expression of gene and protein in the venous bridge wall of diabetic rats increased with time, and were higher than those of normal rats in the same period after operation (P0.05). Conclusion the proliferative remodeling of grafts in diabetic rats is closely related to the high expression of BMP4 in vascular wall, and BMP4 may play an important role in accelerating the failure of venous bridges in high glucose environment.
【作者单位】：四川大学华西医院麻醉科;四川大学华西医院心脏大血管外科;
【基金】：国家自然科学基金(No.81300155,No.81670327)资助
【分类号】：R614

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