PAI-1和vWF在幼鼠以及兒童青少年MS中的表達(dá)與意義的研究
發(fā)布時間：2018-07-26 12:24
【摘要】：一、代謝綜合征幼鼠模型的建立及血管內(nèi)皮功能損傷的檢測 目的：通過高脂高鹽飲食建立代謝綜合征(]metabolic syndrome, MS)幼鼠模型,觀察MS幼鼠腹主動脈的病理變化,并檢測血管內(nèi)皮功能損傷標(biāo)記物--纖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幼鼠腹主動脈血管壁以及血清中的表達(dá)變化,探討MS幼鼠PAI-1和vWF在血管內(nèi)皮功能損傷(endothelial cell dysfunction, ECD)發(fā)生中的作用和意義。 方法：3周SD幼鼠36只,隨機(jī)分為三組,分別給予高脂高鹽飲食、高脂飲食、普通飲食,飼養(yǎng)4周,普通飲食組(NC組)、高脂飲食組(FC組)及高脂高鹽飲食組(FSC組)每組12只。生長至7周末,分別測量體重、體長、腹圍、血壓,禁食12h后取尾靜脈血檢測血總膽固醇(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)及空腹胰島素(Fasting insulin,FINS)水平,處死動物后取腎周內(nèi)臟脂肪測內(nèi)臟脂肪重量,計算三組之間各指標(biāo)是否具有統(tǒng)計學(xué)意義,以確定MS幼鼠模型是否建立成功。模型建立成功后,取幼鼠的腹主動脈,以HE染色法觀察三組動物腹主動脈的病理改變有無差別,采用免疫組化法檢測PAI-1、vWF在三組動物腹主動脈血管壁的表達(dá)以及酶聯(lián)免疫吸附試驗法(enzyme-linked immunosorbent assay, ELISA)檢測三組動物血清中PAI-1、vWF的含量,進(jìn)行統(tǒng)計學(xué)分析,比較PAI-1、vWF在三組動物中的表達(dá)是否具有統(tǒng)計學(xué)意義,以驗證MS幼鼠是否合并ECD。 結(jié)果：1.FSC組幼鼠體重、腹圍、內(nèi)臟脂肪重量顯著高于NC組,差異具有統(tǒng)計學(xué)意義(P0.05),收縮壓(Systolic Blood Prsssure,SBP)、舒張壓(Diastolic Blood Pressure, DBP)、TG、TC水平均顯著高于NC組,HDL水平顯著低于NC組,差異具有統(tǒng)計學(xué)意義(P0.05),FPG、FINS與NC組比較差異具有統(tǒng)計學(xué)意義(P0.05),FSC組幼鼠具備了兒童青少年MS的特征。2.FC組體重、腹圍、內(nèi)臟脂肪重量、FPG、FINS與NC組相比差異存在統(tǒng)計學(xué)意義(P0.05),而血壓、血脂指標(biāo)與NC組比較不具有統(tǒng)計學(xué)意義(P0.05),因而FC組幼鼠只有肥胖、糖代謝異常,不符合MS的診斷標(biāo)準(zhǔn)。3.腹主動脈光鏡檢查顯示：NC、FC組大鼠腹主動脈內(nèi)皮細(xì)胞形態(tài)正常,血管壁結(jié)構(gòu)較完整,而FSC組大鼠腹主動脈內(nèi)皮細(xì)胞少許脫落,肌層細(xì)胞排列紊亂。4.PAI-1、vWF在FSC組大鼠腹主動脈血管壁的免疫組化染色表達(dá)明顯高于NC. FC組,差異具有統(tǒng)計學(xué)意義(P0.05)。5.FSC組血清中PAI-1、vWF含量明顯高于NC、FC組,差異具有統(tǒng)計學(xué)意義(P0.01)。 結(jié)論：1.3周齡SD幼鼠給予高脂高鹽飲食4周后,FSC組幼鼠出現(xiàn)了腹型肥胖、空腹血糖高、高血壓、高TG、低HDL等特征,這些改變與兒童青少年MS的臨床特征相似,說明通過高脂高鹽飲食成功建立了MS幼鼠模型。FC組雖然出現(xiàn)了腹型肥胖、糖代謝異常,但是無高血壓、脂質(zhì)代謝紊亂等特征,不符合MS的診斷標(biāo)準(zhǔn)。2PAI-1、vWF在FSC組幼鼠腹主動脈血管壁以及血清中的表達(dá)均高于FC組及NC組,說明MS幼鼠已有ECD,而FC組中的PAI-1、vWF在腹主動脈血管壁以及血清中的表達(dá)與NC組相比未見明顯差異,表明單純性肥胖幼鼠可能尚未出現(xiàn)ECD。 二、代謝綜合征兒童青少年血PAI-1、vWF與代謝紊亂關(guān)系的研究 目的：通過檢測血管內(nèi)皮功能損傷標(biāo)志物—PAI-1及vWF在肥胖及MS兒童青少年血清中的表達(dá),探討兒童青少年MS與ECD的關(guān)系。方法：收集2011年2月-2012年2月期間于天津醫(yī)科大學(xué)總醫(yī)院兒科內(nèi)分泌門診就診的肥胖兒童病例共60例,其中單純性肥胖30例,包括男16例,女14例,診斷為MS的30例,包括男17例,女13例,并隨機(jī)抽取同年齡健康查體的正常對照兒童30例,包括男15例,女15例。測量身高(H)、體重(W)、腰圍、臀圍、血壓,計算體質(zhì)指數(shù)(BMI),檢測血總膽固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL)、高密度脂蛋白(HDL)、超敏CRP(hsCRP)、糖化血紅蛋白(HbAlc)以及進(jìn)行糖耐量實驗(OGTT)。采用酶聯(lián)免疫吸附法(ELISA)測定所有受試者空腹靜脈血血清PAI-1、vWF的水平。比較三組之間各指標(biāo)的水平,進(jìn)行統(tǒng)計學(xué)分析,對MS組血清PAI-1水平與體格測量參數(shù)及糖脂代謝各指標(biāo)進(jìn)行相關(guān)分析。結(jié)果：1.MS組的PAI-1及vWF水平顯著高于對照組,差異具有統(tǒng)計學(xué)意義(P0.05)。2.單純性肥胖組血清PAI-1水平顯著高于對照組,差異有統(tǒng)計學(xué)意義(P=0.003),血清vWF與對照組比較差別無統(tǒng)計學(xué)意義(P=0.556)。3.相關(guān)性分析顯示PAI-1濃度與體重、BMI、腰圍、TC、TG、LDL、FPG、FINS、SBP、DBP、hsCRP呈正相關(guān)(P0.05),與HDL呈負(fù)相關(guān)(P0.05),與臀圍、腰臀比、HbAlc、2hPG無相關(guān)性(P0.05),進(jìn)一步多元逐步回歸分析顯示FINS、TC,TG可獨立影響PAI-1的濃度。結(jié)論：1.兒童青少年MS合并ECD。PAI-1及vWF可以作為臨床中預(yù)測早期血管改變的指標(biāo)。2.兒童青少年MS存在的ECD可能與肥胖、血脂、血糖、血壓均相關(guān),而FINS、TC、TG可能是ECD的獨立危險因素。
[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.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R725.8
本文編號：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.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R725.8
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