神经调节蛋白1在糖尿病大鼠静脉桥中的表达变化及意义

发布时间：2018-06-12 01:24

本文选题：糖尿病 + 神经调节蛋白1　；参考：《广西医科大学》2015年硕士论文

【摘要】：研究背景和目的：研究发现至少65%糖尿病(DM)患者死亡与冠状动脉动脉硬化性心脏病有关。冠状动脉血管旁路移植手术(CABG)是治疗终末期冠脉严重病变的重要手段。尽管多种动脉移植物相继被应用于CABG中,但因其来源不足在临床应用中仍受到很大限制。迄今大隐静脉仍是最为常用的血管桥。而应用大隐静脉桥进行CABG合并糖尿病术后远期心肌梗死和死亡率显著上升。DM静脉桥术后内膜增生导致再狭窄的原因及作用机制尚未可知,同时亦无理想的防治方法。最近研究表明NRG-1在糖尿病心血管疾病表达明显下调,而且NRG-1能保护血管内皮细胞和抑制平滑肌增殖、迁移,亦能减少氧化应激。因此探讨NRG-1是否参与DM静脉桥再狭窄的过程是很有必要的。本研究旨在动物上建立DM静脉桥模型,探究静脉桥在DM环境下内膜增厚情况以及NRG-1表达变化,有望为DM患者血管桥再狭窄的预防及治疗提供新的途径。研究方法：1、随机选用8-10周龄雄性(Sprague-Dawley rats)SD大鼠50只,体重200-220g,给予标准大鼠饲料,随机分为两组：糖尿病组(DM组,n=25)和正常对照组(NC组,n=25),DM组给予大剂量STZ(55mg/kg)腹腔注射建立糖尿病大鼠模型；NC组给予等体积生理盐水腹腔注射。3天后尾静脉采血测空腹血糖16.7mmol/L的大鼠纳入糖尿病模型组。2、选取造模成功的DM大鼠(DM组)与正常大鼠(NC组)随机分为3组,分别为DM大鼠0周(DM-OW、2周(DM-2W,n=6)、4周(DM-4W,n=6)及正常组0周(NC-OW,n=6)、2周(NC-2W,n=6)、4周(NC-4W,n=6), STZ注射1周后,采用改良cuff管法对所有大鼠建立自体颈外静脉移植颈总动脉模型。3、分别取两组0周、2周、4周的血管桥行病理检测,测量两组血管桥管壁的内、中膜厚度并比较二者的差别。免疫组化检测PCNA和NRG1的表达情况,同时利用Western blot测定NRG1的相对表达量。结果：1、25只大鼠糖尿病造模成功24只,造模成功率为96%(24/25),其中有1只在注射药物后第二天死亡,死亡率为4%(1/25)考虑STZ中毒死亡所致。2、49只大鼠行自体颈外静脉移植至颈总动脉模型,建模成功45例,成功率为91.84%(45/49),其中DM组22例,NC组23例；其死亡率8.16%(4/49)。3、大鼠自体颈外静脉移植术前糖尿病组静脉桥壁内膜厚度与对照组相比(4.54±0.39μm vs 4.59±0.6μm,P0.05),没有统计学差异；而术后2周(50.68±2.34μm vs 45.95±3.1μm, P0.05),有统计学差异；术后4W (80.86±8.72μm vs 60.7±6.31μm, P0.01),有统计学差异。4、DM组和NC组术前静脉桥中平滑肌细胞PCNA几乎无阳性表达。移植术后2周血管平滑肌细胞PCNA阳性表达明显增多并达高峰,以内、中膜为主,4周阳性率明显降低,与对照组相比,在术后2周(51.99±5.23 vs 30.5±5.16, P0.01)差异有统计学意义,在4周(18.85±3.20 vs 15.14±1.69, P0.05)差异仍有统计学意义。5、NRG1主要在平滑肌细胞的细胞质和细胞膜为主。术后阳性率表达有所下降,以DM比较显著；与对照组相比,0周时(85.01±1.87 vs 84.64 ±1.09, P0.05)无统计学意义,术后2周(72.12±6.30 vs 28.58±1.94, P 0.05)以及到4周时(34.53±2.07 vs 7.89±1.37, P0.01)差异均有统计学意义；PCNA与NRG1阳性率经相关性统计分析结果示：NC组相关系数r=-0.21,P=0.4030.05,负相关无统计学意义,DM组相关系数r=-0.59,P=0.0.010.05,负相关有统计学意义。结论：1、大鼠自体移植静脉手术后早期开始出现新生内膜,逐渐增厚,主要是VSMC增殖、迁移所致2、自体静脉移植术后同一时间点,与对照组相比,糖尿病内膜增厚明显；3、自体静脉移植术后,NRG1表达下调显著以致PCNA高表达,可能是糖尿病组VSMC增殖、内膜增厚明显的关键因素之一；4、NRG1可能成为糖尿病再狭窄治疗的新的靶点。
[Abstract]:Background and purpose: the study found that at least 65% patients with diabetes (DM) died of coronary arteriosclerotic heart disease. Coronary artery bypass grafting (CABG) is an important means for the treatment of severe end-stage coronary lesions. Although multiple arterial graft has been applied to CABG successively, it is clinically used because of its lack of origin. So far the great saphenous vein is still the most commonly used vascular bridge. The long term myocardial infarction and mortality after the use of the great saphenous vein bridge for CABG combined with diabetes has significantly increased the cause and mechanism of intimal hyperplasia after.DM bridge, and there is no ideal method for prevention and treatment. It shows that the expression of NRG-1 in diabetic cardiovascular disease is obviously down-regulated, and NRG-1 can protect vascular endothelial cells and inhibit the proliferation of smooth muscle, migration, and reduce oxidative stress. Therefore, it is necessary to explore whether NRG-1 is involved in the process of DM vein bridge restenosis. This study aims to establish a DM vein bridge model and explore the vein bridge in DM. The intimal thickening and NRG-1 expression changes in the environment are expected to provide a new way for the prevention and treatment of restenosis of vascular bridges in DM patients. 1, 50 rats were randomly selected for 8-10 weeks male (Sprague-Dawley rats) SD rats and weighed 200-220g, and were randomly divided into two groups: diabetes group (DM group, n=25) and normal group. The control group (group NC, n=25), group DM was given a large dose of STZ (55mg/kg) intraperitoneally to establish the diabetic rat model, and the NC group was given.2 in the diabetic rat model group with equal volume of normal saline injected with the tail vein of.3 after.3 days, and the DM rats (DM group) and normal rats (NC group) were randomly divided into 3 groups. 0 weeks (DM-OW, 2 weeks (DM-2W, n=6), 4 weeks (DM-4W, n=6) and normal group 0 weeks (NC-OW, n=6), 2 weeks (NC-2W, n=6), 4 weeks (NC-4W, n=6), after 1 weeks of injection, all rats were established by modified jugular vein graft common carotid artery model, and two groups of 0 weeks, 2 weeks, 4 weeks of vascular bridge examination, respectively, to measure two group of blood. The thickness of the tube wall and the difference in the thickness of the middle membrane were compared with those of the two. The expression of PCNA and NRG1 was detected by immunohistochemistry and the relative expression of NRG1 was measured by Western blot. Results: 24 rats were successfully established with diabetes, and the success rate of the model was 96% (24/25), of which 1 were killed second days after the injection and the mortality rate was 4% (1/25). The model of autologous external jugular vein graft to common carotid artery was considered in.2,49 rats with STZ poisoning death. 45 cases were successfully modeled, the success rate was 91.84% (45/49), including 22 cases in group DM and 23 in NC group; the mortality rate was 8.16% (4/49).3, and the intima thickness of vein bridge wall in the diabetic group was compared with the control group (4.54 + 0.39, m vs 4). .59 + 0.6 m, P0.05), without statistical difference, but 2 weeks after operation (50.68 + 2.34 Mu vs 45.95 + 3.1 m, P0.05), there were statistical differences; 4W (80.86 + 8.72 micron vs 60.7 + 6.31 mu m, P0.01) was statistically different after operation. The positive expression of CNA was significantly increased and reached its peak. The positive rate of the middle membrane was less than that of the control group. Compared with the control group, the difference was statistically significant at the 2 week (51.99 + 5.23 vs 30.5 + 5.16, P0.01) after the operation. The difference was still statistically significant at 4 weeks (18.85 + 3.20 vs 15.14 +, P0.05), and NRG1 mainly in the cytoplasm and cell of smooth muscle cells. The expression of membrane was dominant. The expression of positive rate decreased after operation. Compared with the control group, there was no statistical significance at 0 weeks (85.01 + 1.87 vs 84.64 + 1.09, P0.05), 2 weeks after operation (72.12 + 6.30 vs 28.58 + 1.94, P 0.05) and 1.87 weeks (34.53 + vs, P0.01) differences were statistically significant; PCNA and NRG1 positive rates were in phase The correlation coefficient of the NC group was r=-0.21, P=0.4030.05, and the negative correlation was not statistically significant. The correlation coefficient of group DM was r=-0.59, P=0.0.010.05, and the negative correlation was statistically significant. Conclusion: 1, the neointima began to appear in the early stage of autologous vein transplantation in rats, gradually thickening, mainly the proliferation of VSMC, the migration of 2, autogenous vein migration. At the same time after implantation, the thickening of the intima of diabetes was obvious compared with the control group. 3, after autologous vein transplantation, the expression of NRG1 was significantly reduced so that the high expression of PCNA could be one of the key factors for the proliferation of VSMC in the diabetic group and the thickening of the intima, and 4, NRG1 may be a new target for the treatment of diabetes restenosis.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R587.1

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