vWF、PAI-1在糖尿病大鼠肾组织的表达及丹红注射液、二甲双胍的干预研究

发布时间：2018-06-17 06:42

本文选题：糖尿病肾病 + vWF　；参考：《河北医科大学》2014年硕士论文

【摘要】：目的：糖尿病是一种全身的慢性代谢性疾病，其发病率逐年增高。糖尿病肾病（DN)是糖尿病最常见和严重的微血管并发症，也是导致终末期肾脏病的一个重要原因。DN的发生发展是多因素综合作用的结果,与糖代谢紊乱、血流动力学、细胞因子、氧化应激、炎症反应、血液流变学等因素密切相关。血液流变学改变是糖尿病时微循环障碍的主要原因,在DN发展过程中起重要作用。血管性假血友病因子（vWF）、纤溶酶原激活物抑制剂-1（PAI-1）能反映机体的凝血、纤溶功能，参与DN的发生、发展。vWF是血管内皮细胞损伤的标志物。vWF水平升高促进了血液的高凝状态。PAI-1是纤溶系统的主要调控因子,能够抑制纤溶活性。丹红注射液的主要功能为活血化瘀，二甲双胍是一种被广泛应用的有效安全的降糖药，，同时具有调节血脂,降低血小板聚集、改善血流变的作用。丹红注射液、二甲双胍可能能够延缓DN的进展，但其具体机制尚待进一步研究。本研究观察丹红注射液、二甲双胍干预前后糖尿病大鼠血清vWF水平及肾组织中vWF、PAI-1表达的变化，进一步探讨DN发病机制及丹红注射液、二甲双胍的肾脏保护作用。方法:48只雄性SD大鼠随机分为2组:正常对照组（A组）10只、实验组38只。实验组大鼠均按55mg/kg剂量一次性腹腔注射链脲佐菌素(STZ)，72小时后测血糖值＞16.7mmol/L，持续三天，作为糖尿病大鼠模型建立的标准。把实验组大鼠随机分为三组：糖尿病组（B组）14只、丹红干预组（C组）12只、二甲双胍干预组（D组）12只。C组按2ml/kg·d剂量腹腔注射丹红注射液，D组给予二甲双胍300mg/kg·d灌胃给药，A和B组分别给予腹腔注射相应剂量的生理盐水。干预为期8周。于第8周末，各组大鼠收集血液、尿液及肾组织标本。然后检测血糖（BG)、总胆固醇（TC）、甘油三酯（TG）、低密度脂蛋白胆固醇（LDL-C）、血肌酐（Scr）、尿素氮（BUN）、24小时尿白蛋白定量。酶联免疫分析法（ELISA）测定各组大鼠血清vWF的浓度。肾组织进行苏木素-伊红（HE)染色在光镜观察肾脏形态学变化。用免疫组化的方法检测各组大鼠肾脏vWF的表达，应用图像分析系统进行分析。应用实时荧光定量PCR技术检测肾组织vWF、PAI-1mRNA的表达情况。应用SPSS16.0统计学软件处理实验数据,采用单因素方差分析、非参数检验实验方法，以P0.05为差异有统计学意义。结果： 18周后，B组大鼠BG、TC、TG、LDL-C、Scr、BUN、24小时尿白蛋白定量、肾脏肥大指数明显高于A组(P0.01)，应用丹红注射液、二甲双胍干预8周后，干预组大鼠TC、TG、LDL-C、Scr、BUN、24小时尿白蛋白定量、肾脏肥大指数较B组明显减低(P0.05),D组BG较B组相比明显降低(P0.01)。 2血清vWF:B组血清vWF的浓度较A组明显升高(P0.01)，而C组、D组较B组水平有所下降(P0.01)，差异有统计学意义。 3形态学检查：糖尿病大鼠出现病理改变，B组大鼠可见肾小球平均截面积明显增大，系膜基质轻度增多，部分肾小管上皮细胞萎缩，管腔扩张。C组、D组大鼠肾脏病理改变减轻。 4肾组织vWF蛋白的表达：免疫组化结果提示B组肾组织vWF蛋白表达明显高于A组(P0.01)；C组、D组大鼠肾脏vWF的表达较B组明显减少(P0.01)，但仍多于A组。 5实时荧光定量PCR结果提示：与A组相比，B组vWF、PAI-1mRNA的表达明显增加(P0.01)，而C组、D组vWF、PAI-1mRNA表达较B组有所减少(P0.05)。B组、C组、D组大鼠肾组织vWFmRNA相对表达量分别是A组的5.09倍、3.45倍、3.19倍。B组、C组、D组大鼠肾组织PAI-1mRNA相对表达量分别是A组的4.31倍、2.70倍、2.98倍。结论： 1实验性1型糖尿病大鼠血清vWF水平及肾脏vWF、PAI-1表达增加，提示vWF、PAI-1与糖尿病肾病的发生、发展关系密切。 2丹红注射液、二甲双胍通过调节血脂、降低尿白蛋白、降低vWF和PAI-1的表达，来发挥肾脏保护作用。
[Abstract]:Objective : Diabetic nephropathy ( DN ) is one of the most common and serious microvascular complications of diabetes mellitus .

Methods : Forty - eight male SD rats were randomly divided into two groups : normal control group ( group A ) and experimental group ( n = 38 ) . The rats were divided into three groups : diabetic group ( group B ) , total cholesterol ( TC ) , triglyceride ( TG ) , low density lipoprotein cholesterol ( LDL - C ) , serum creatinine ( TC ) , urea nitrogen ( BUN ) and urine albumin .

Results :

After 18 weeks , BG , TC , TG , LDL - C , BUN , 24h urine albumin and renal hypertrophy index in group B were significantly higher than those in group A ( P0.01 ) . After 8 weeks of intervention with Danhong injection and metformin , the levels of TC , TG , LDL - C , BUN , 24h urine albumin were significantly lower in the intervention group than in group B ( P0.05 ) , while those in group D were significantly lower than those in group B ( P0.01 ) .

There was a significant difference in the levels of the serum levels in the serum from the group A and the group D ( P0.01 ) , and the difference was statistically significant .

The pathological changes in diabetic rats showed that the mean cross - sectional area of glomerulus in group B was obviously increased , the mesangial matrix was slightly increased , partial renal tubular epithelial cells were atrophied , the lumen was dilated , and the pathological changes of renal pathology in group C and D were reduced .

The results showed that there was a significant difference in the levels of plasma protein in renal tissue in group B than in group A ( P0.01 ) .
In group C and D rats , the expression of VWF decreased significantly compared with group B ( P0.01 ) , but there were more than group A .

Results : Compared with group A , the expression of PAI - 1 mRNA in group B , group C and D was 5.09 times , 3.45 times and 3.19 times as compared with group A , and the relative expression of PAI - 1 mRNA in group B , group C and D was 4.31 times , 2.70 times and 2.98 times of group A , respectively .

Conclusion :

Results The levels of serum levels and the level of PAI - 1 in the serum of experimental type 1 diabetic rats were increased . The results suggested that the occurrence and development of the levels of the levels of the levels of the levels of the levels of the levels of the levels of the plasma levels in the rat were closely related to the occurrence and development of diabetic nephropathy .

2 Danhong injection , metformin can play a role in kidney protection by regulating blood lipid , reducing urinary albumin , decreasing the expression of PAI - 1 .
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R587.2;R692

【参考文献】