PAI-1和vWF在幼鼠以及儿童青少年MS中的表达与意义的研究
发布时间：2018-07-26 12:24
【摘要】：一、代谢综合征幼鼠模型的建立及血管内皮功能损伤的检测 目的：通过高脂高盐饮食建立代谢综合征(]metabolic syndrome, MS)幼鼠模型,观察MS幼鼠腹主动脉的病理变化,并检测血管内皮功能损伤标记物--纤溶酶原激活物抑制物-1(plasminogen activator inhibitor-1, PAI-1).血管性血友病因子(von Willebrand Factor, vWF)在MS幼鼠腹主动脉血管壁以及血清中的表达变化,探讨MS幼鼠PAI-1和vWF在血管内皮功能损伤(endothelial cell dysfunction, ECD)发生中的作用和意义。 方法：3周SD幼鼠36只,随机分为三组,分别给予高脂高盐饮食、高脂饮食、普通饮食,饲养4周,普通饮食组(NC组)、高脂饮食组(FC组)及高脂高盐饮食组(FSC组)每组12只。生长至7周末,分别测量体重、体长、腹围、血压,禁食12h后取尾静脉血检测血总胆固醇(Cholesterol, TC)、甘油三酯(Triglyceride, TG)、高密度脂蛋白(High-density lipoprotein, HDL)、空腹血糖(Fasting blood glucose, FPG)及空腹胰岛素(Fasting insulin,FINS)水平,处死动物后取肾周内脏脂肪测内脏脂肪重量,计算三组之间各指标是否具有统计学意义,以确定MS幼鼠模型是否建立成功。模型建立成功后,取幼鼠的腹主动脉,以HE染色法观察三组动物腹主动脉的病理改变有无差别,采用免疫组化法检测PAI-1、vWF在三组动物腹主动脉血管壁的表达以及酶联免疫吸附试验法(enzyme-linked immunosorbent assay, ELISA)检测三组动物血清中PAI-1、vWF的含量,进行统计学分析,比较PAI-1、vWF在三组动物中的表达是否具有统计学意义,以验证MS幼鼠是否合并ECD。 结果：1.FSC组幼鼠体重、腹围、内脏脂肪重量显著高于NC组,差异具有统计学意义(P0.05),收缩压(Systolic Blood Prsssure,SBP)、舒张压(Diastolic Blood Pressure, DBP)、TG、TC水平均显著高于NC组,HDL水平显著低于NC组,差异具有统计学意义(P0.05),FPG、FINS与NC组比较差异具有统计学意义(P0.05),FSC组幼鼠具备了儿童青少年MS的特征。2.FC组体重、腹围、内脏脂肪重量、FPG、FINS与NC组相比差异存在统计学意义(P0.05),而血压、血脂指标与NC组比较不具有统计学意义(P0.05),因而FC组幼鼠只有肥胖、糖代谢异常,不符合MS的诊断标准。3.腹主动脉光镜检查显示：NC、FC组大鼠腹主动脉内皮细胞形态正常,血管壁结构较完整,而FSC组大鼠腹主动脉内皮细胞少许脱落,肌层细胞排列紊乱。4.PAI-1、vWF在FSC组大鼠腹主动脉血管壁的免疫组化染色表达明显高于NC. FC组,差异具有统计学意义(P0.05)。5.FSC组血清中PAI-1、vWF含量明显高于NC、FC组,差异具有统计学意义(P0.01)。 结论：1.3周龄SD幼鼠给予高脂高盐饮食4周后,FSC组幼鼠出现了腹型肥胖、空腹血糖高、高血压、高TG、低HDL等特征,这些改变与儿童青少年MS的临床特征相似,说明通过高脂高盐饮食成功建立了MS幼鼠模型。FC组虽然出现了腹型肥胖、糖代谢异常,但是无高血压、脂质代谢紊乱等特征,不符合MS的诊断标准。2PAI-1、vWF在FSC组幼鼠腹主动脉血管壁以及血清中的表达均高于FC组及NC组,说明MS幼鼠已有ECD,而FC组中的PAI-1、vWF在腹主动脉血管壁以及血清中的表达与NC组相比未见明显差异,表明单纯性肥胖幼鼠可能尚未出现ECD。 二、代谢综合征儿童青少年血PAI-1、vWF与代谢紊乱关系的研究 目的：通过检测血管内皮功能损伤标志物—PAI-1及vWF在肥胖及MS儿童青少年血清中的表达,探讨儿童青少年MS与ECD的关系。方法：收集2011年2月-2012年2月期间于天津医科大学总医院儿科内分泌门诊就诊的肥胖儿童病例共60例,其中单纯性肥胖30例,包括男16例,女14例,诊断为MS的30例,包括男17例,女13例,并随机抽取同年龄健康查体的正常对照儿童30例,包括男15例,女15例。测量身高(H)、体重(W)、腰围、臀围、血压,计算体质指数(BMI),检测血总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL)、高密度脂蛋白(HDL)、超敏CRP(hsCRP)、糖化血红蛋白(HbAlc)以及进行糖耐量实验(OGTT)。采用酶联免疫吸附法(ELISA)测定所有受试者空腹静脉血血清PAI-1、vWF的水平。比较三组之间各指标的水平,进行统计学分析,对MS组血清PAI-1水平与体格测量参数及糖脂代谢各指标进行相关分析。结果：1.MS组的PAI-1及vWF水平显著高于对照组,差异具有统计学意义(P0.05)。2.单纯性肥胖组血清PAI-1水平显著高于对照组,差异有统计学意义(P=0.003),血清vWF与对照组比较差别无统计学意义(P=0.556)。3.相关性分析显示PAI-1浓度与体重、BMI、腰围、TC、TG、LDL、FPG、FINS、SBP、DBP、hsCRP呈正相关(P0.05),与HDL呈负相关(P0.05),与臀围、腰臀比、HbAlc、2hPG无相关性(P0.05),进一步多元逐步回归分析显示FINS、TC,TG可独立影响PAI-1的浓度。结论：1.儿童青少年MS合并ECD。PAI-1及vWF可以作为临床中预测早期血管改变的指标。2.儿童青少年MS存在的ECD可能与肥胖、血脂、血糖、血压均相关,而FINS、TC、TG可能是ECD的独立危险因素。
[Abstract]:Establishment of a metabolic syndrome rat model and detection of endothelial dysfunction
Objective: to establish a model of]metabolic syndrome (MS) by high fat and high salt diet, and to observe the pathological changes of abdominal aorta in young MS rats, and to detect the vascular endothelial dysfunction markers, -1 (plasminogen activator inhibitor-1, PAI-1), and vascular hemophilia factor (von Willebrand Factor). VWF) changes in the vascular wall of abdominal aorta and serum in young MS rats, and explore the role and significance of PAI-1 and vWF in MS young rats in the occurrence of vascular endothelial dysfunction (endothelial cell dysfunction, ECD).
Methods: 36 SD young rats were randomly divided into three groups, which were randomly divided into three groups, which were given high fat and high salt diet, high fat diet, diet for 4 weeks, ordinary diet group (group NC), high fat diet group (group FC) and high fat and high salt diet group (group FSC) 12. The body weight, body length, abdominal circumference, blood pressure, and 12h after fasting were measured to measure the total blood total. Cholesterol (TC), triglyceride (Triglyceride, TG), high density lipoprotein (High-density lipoprotein, HDL), fasting blood glucose (Fasting blood glucose, FPG) and fasting insulin (Fasting) levels were executed to measure the visceral fat weight of the perinenal visceral fat, and the statistics between the three groups were calculated. To determine whether the model of MS young rats was established successfully. After the model was successfully established, the abdominal aorta of the young rats was taken. The pathological changes of abdominal aorta in the three groups were observed by HE staining. The expression of PAI-1, the expression of vWF in the abdominal aorta wall of the three groups of animals and the enzyme linked immunosorbent assay (enzyme-linked) were detected by the immunohistochemical method (enzyme-linked Immunosorbent assay, ELISA) detected the content of PAI-1 and vWF in the serum of three groups of animals. Statistical analysis was carried out to compare the statistical significance of PAI-1, vWF in the three groups of animals, in order to verify whether MS young rats were combined with ECD..
Results: the weight, abdominal circumference and visceral fat weight of young rats in group 1.FSC were significantly higher than that in group NC (P0.05), systolic pressure (Systolic Blood Prsssure, SBP), diastolic pressure (Diastolic Blood Pressure, DBP), TG, and DBP were significantly higher than those in the group, and the difference was statistically significant. The difference was statistically significant (P0.05). The young rats in group FSC had the body weight of MS in children and adolescents, abdominal circumference, visceral fat weight, FPG, FINS and NC group had statistical significance (P0.05), while blood pressure, blood lipid index and NC group had no statistical significance (P0.05), so the young rats in FC group had only obesity and abnormal glucose metabolism. The.3. abdominal aortic optical microscopy, which did not conform to the diagnostic standard of MS, showed that the abdominal aorta endothelial cells in the group FC rats were normal and the vascular wall structure was more complete, while the abdominal aorta endothelial cells in the group FSC rats were little off, the myometrium cells were arranged in disorder.4.PAI-1, and the immunohistochemical staining of vWF in the abdominal aorta wall of FSC group rats was significantly higher. In NC. FC group, the difference was statistically significant (P0.05). The levels of PAI-1 and vWF in serum of.5.FSC group were significantly higher than those of NC, FC group, the difference was statistically significant (P0.01).
Conclusion: after 4 weeks of high fat and high salt diet for 1.3 week old SD young rats, the young rats in group FSC had abdominal obesity, high blood glucose, high blood pressure, high TG, low HDL and so on. These changes were similar to the clinical characteristics of MS in children and adolescents. It was demonstrated that abdominal obesity and glucose metabolism were found in the.FC group of MS young rats through high fat and high salt diet. However, there was no hypertension and lipid metabolism disorder, which did not conform to the diagnostic standard of MS.2PAI-1. The expression of vWF in the abdominal aorta wall and serum in the FSC group was higher than that in the FC group and the NC group, indicating that the MS young rats had ECD, while the PAI-1 in the FC group and the expression in the serum of the abdominal aorta were not significantly different from those in the NC group. It shows that simple obese rats may not have ECD.
Two, the relationship between PAI-1, vWF and metabolic disorders in children and adolescents with metabolic syndrome
Objective: To investigate the relationship between MS and ECD in children and adolescents by detecting the expression of PAI-1 and vWF in the serum of obese and MS children and adolescents. Methods: a total of 60 obese children were collected in the pediatric endocrinology clinic of General Hospital Affiliated to Tianjin Medical University during February February 2011. 30 cases of sexual obesity, including 16 male and 14 female, 30 cases of MS, were diagnosed as male 17 and 13, and 30 cases of normal control children were randomly selected, including 15 men and 15 women. The height (H), weight (W), waist circumference, hip circumference, blood pressure, body mass index (BMI), blood total cholesterol (TC), triglyceride (TG), low density fat eggs were detected. White (LDL), high density lipoprotein (HDL), hypersensitive CRP (hsCRP), glycated hemoglobin (HbAlc) and glucose tolerance test (OGTT). Enzyme linked immunosorbent assay (ELISA) was used to determine the level of PAI-1 and vWF in all subjects' fasting venous blood serum. The level of each index between the three groups was compared, and the PAI-1 level and physique of the MS group were analyzed. Results: the PAI-1 and vWF levels in the 1.MS group were significantly higher than those in the control group. The difference was statistically significant (P0.05) in.2. simple obesity group, the serum PAI-1 level was significantly higher than that of the control group (P=0.003), and there was no statistical difference between the serum vWF and the control group (P=0.55). 6).3. correlation analysis showed that PAI-1 concentration and weight, BMI, waist circumference, TC, TG, LDL, FPG, FINS, SBP, DBP, hsCRP were positively correlated (P0.05). ECD.PAI-1 and vWF can be used as an indicator of early vascular changes in clinical prediction.2. children and adolescents MS ECD may be associated with obesity, blood lipids, blood sugar, blood pressure, and FINS, TC, TG may be an independent risk factor for ECD.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R725.8
本文编号：2146005
[Abstract]:Establishment of a metabolic syndrome rat model and detection of endothelial dysfunction
Objective: to establish a model of]metabolic syndrome (MS) by high fat and high salt diet, and to observe the pathological changes of abdominal aorta in young MS rats, and to detect the vascular endothelial dysfunction markers, -1 (plasminogen activator inhibitor-1, PAI-1), and vascular hemophilia factor (von Willebrand Factor). VWF) changes in the vascular wall of abdominal aorta and serum in young MS rats, and explore the role and significance of PAI-1 and vWF in MS young rats in the occurrence of vascular endothelial dysfunction (endothelial cell dysfunction, ECD).
Methods: 36 SD young rats were randomly divided into three groups, which were randomly divided into three groups, which were given high fat and high salt diet, high fat diet, diet for 4 weeks, ordinary diet group (group NC), high fat diet group (group FC) and high fat and high salt diet group (group FSC) 12. The body weight, body length, abdominal circumference, blood pressure, and 12h after fasting were measured to measure the total blood total. Cholesterol (TC), triglyceride (Triglyceride, TG), high density lipoprotein (High-density lipoprotein, HDL), fasting blood glucose (Fasting blood glucose, FPG) and fasting insulin (Fasting) levels were executed to measure the visceral fat weight of the perinenal visceral fat, and the statistics between the three groups were calculated. To determine whether the model of MS young rats was established successfully. After the model was successfully established, the abdominal aorta of the young rats was taken. The pathological changes of abdominal aorta in the three groups were observed by HE staining. The expression of PAI-1, the expression of vWF in the abdominal aorta wall of the three groups of animals and the enzyme linked immunosorbent assay (enzyme-linked) were detected by the immunohistochemical method (enzyme-linked Immunosorbent assay, ELISA) detected the content of PAI-1 and vWF in the serum of three groups of animals. Statistical analysis was carried out to compare the statistical significance of PAI-1, vWF in the three groups of animals, in order to verify whether MS young rats were combined with ECD..
Results: the weight, abdominal circumference and visceral fat weight of young rats in group 1.FSC were significantly higher than that in group NC (P0.05), systolic pressure (Systolic Blood Prsssure, SBP), diastolic pressure (Diastolic Blood Pressure, DBP), TG, and DBP were significantly higher than those in the group, and the difference was statistically significant. The difference was statistically significant (P0.05). The young rats in group FSC had the body weight of MS in children and adolescents, abdominal circumference, visceral fat weight, FPG, FINS and NC group had statistical significance (P0.05), while blood pressure, blood lipid index and NC group had no statistical significance (P0.05), so the young rats in FC group had only obesity and abnormal glucose metabolism. The.3. abdominal aortic optical microscopy, which did not conform to the diagnostic standard of MS, showed that the abdominal aorta endothelial cells in the group FC rats were normal and the vascular wall structure was more complete, while the abdominal aorta endothelial cells in the group FSC rats were little off, the myometrium cells were arranged in disorder.4.PAI-1, and the immunohistochemical staining of vWF in the abdominal aorta wall of FSC group rats was significantly higher. In NC. FC group, the difference was statistically significant (P0.05). The levels of PAI-1 and vWF in serum of.5.FSC group were significantly higher than those of NC, FC group, the difference was statistically significant (P0.01).
Conclusion: after 4 weeks of high fat and high salt diet for 1.3 week old SD young rats, the young rats in group FSC had abdominal obesity, high blood glucose, high blood pressure, high TG, low HDL and so on. These changes were similar to the clinical characteristics of MS in children and adolescents. It was demonstrated that abdominal obesity and glucose metabolism were found in the.FC group of MS young rats through high fat and high salt diet. However, there was no hypertension and lipid metabolism disorder, which did not conform to the diagnostic standard of MS.2PAI-1. The expression of vWF in the abdominal aorta wall and serum in the FSC group was higher than that in the FC group and the NC group, indicating that the MS young rats had ECD, while the PAI-1 in the FC group and the expression in the serum of the abdominal aorta were not significantly different from those in the NC group. It shows that simple obese rats may not have ECD.
Two, the relationship between PAI-1, vWF and metabolic disorders in children and adolescents with metabolic syndrome
Objective: To investigate the relationship between MS and ECD in children and adolescents by detecting the expression of PAI-1 and vWF in the serum of obese and MS children and adolescents. Methods: a total of 60 obese children were collected in the pediatric endocrinology clinic of General Hospital Affiliated to Tianjin Medical University during February February 2011. 30 cases of sexual obesity, including 16 male and 14 female, 30 cases of MS, were diagnosed as male 17 and 13, and 30 cases of normal control children were randomly selected, including 15 men and 15 women. The height (H), weight (W), waist circumference, hip circumference, blood pressure, body mass index (BMI), blood total cholesterol (TC), triglyceride (TG), low density fat eggs were detected. White (LDL), high density lipoprotein (HDL), hypersensitive CRP (hsCRP), glycated hemoglobin (HbAlc) and glucose tolerance test (OGTT). Enzyme linked immunosorbent assay (ELISA) was used to determine the level of PAI-1 and vWF in all subjects' fasting venous blood serum. The level of each index between the three groups was compared, and the PAI-1 level and physique of the MS group were analyzed. Results: the PAI-1 and vWF levels in the 1.MS group were significantly higher than those in the control group. The difference was statistically significant (P0.05) in.2. simple obesity group, the serum PAI-1 level was significantly higher than that of the control group (P=0.003), and there was no statistical difference between the serum vWF and the control group (P=0.55). 6).3. correlation analysis showed that PAI-1 concentration and weight, BMI, waist circumference, TC, TG, LDL, FPG, FINS, SBP, DBP, hsCRP were positively correlated (P0.05). ECD.PAI-1 and vWF can be used as an indicator of early vascular changes in clinical prediction.2. children and adolescents MS ECD may be associated with obesity, blood lipids, blood sugar, blood pressure, and FINS, TC, TG may be an independent risk factor for ECD.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R725.8
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