家兔动脉粥样硬化模型中尾加压素Ⅱ血浆含量变化
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摘要
前言
尾加压素Ⅱ(urotensinⅡ, UTⅡ)最早从鱼尾部下垂体中提取出的神经肽,是目前所知的体内最强的缩血管活性肽。其受体广泛分布于哺乳动物的心脏、肾脏、神经组织、血管平滑肌组织、甚至于动脉粥样硬化斑块中。UTⅡ在动脉粥样硬化中发生发展中扮演中重要角色。本实验中我们通过复制家兔动脉粥样硬化模型,观察从正常家兔直至致病过程中尾加压素动态改变,及其与血脂变化,炎症因子含量的相关性,探讨尾加压素在致动脉粥样硬化中的重要意义。
目的
研究动脉粥样硬化模型中UTⅡ血浆含量的动态变化,及其与血脂蛋白炎症因子的相关性分析。
材料方法
1.材料
健康日本大耳白家兔40只,4月龄,雌雄不限,购自沈阳市药科大学,合格证号：SCXK(辽)2009-0002,体重2.0-2.5Kg。
2.高脂饲料的配制
基础饲料中加入胆固醇(国药化学试剂有限公司,上海产,分析纯)、猪油(市售)、蛋黄粉(市售),三种成分按0.6%,3.3%,8.3%配比制成,称为高脂饮食。
3.实验动物处理
随机将家兔分为2组,对照组20只,实验组20只。单笼饲养,自由摄取饮水。实验组家兔饲喂高脂饲料,对照组继续饲喂基础兔饲料。每日每只家兔进食150g,实验组保证每日每只家兔进食胆固醇1.0g,猪油5.0g,蛋黄粉12.5g。总共喂养12周。
4.生化项目检测
于实验的0周、3周、6周、9周、12周抽血留取血浆标本,测定血清TC、HDLC、LDLC水平；应用酶联免疫法测定UTⅡ、IL-6水平。同时进行UTⅡ与血脂蛋白、炎症因子相关性分析。
5.统计学分析
应用SPSS13.0统计学软件,组间比较采用方差分析。
实验结果
1.家兔体重变化,血脂含量变化
喂养12周后家兔体重均增加,但两组间无差异(P>0.05)。实验组家兔血脂水平总体呈升高趋势。6周时,与对照组相比,实验组家兔血脂水平均升高,差别有统计学意义(P<0.05),12周实验组家兔与6周实验组家兔比较,TC和LDLC差别显著(P均<0.01)。
2.家兔血浆炎症因子UTⅡ及IL-6含量的变化
高脂饮食喂养的家兔在3周时血浆中UTⅡ含量开始下降,第12周较对照组及第6周可见其含量明显减少,有显著差异(P均<0.01)。炎症因子IL-6含量在6周时开始升高,但差别无统计学意义(P>0.05),9周及12周可见炎症因子含量明显增高,与对照组及6周实验组有显著差别(P均<0.01)
3.UTⅡ与TC、IL-6相关性分析
家兔动脉粥样硬化模型中UTⅡ与TC血清中含量呈明显负相关, r=-0.99,P=0.01,直线回归方程Y=0.211-0.003X。UTⅡ与IL-6血清中含量呈明显负相关,r=-0.95,P=0.04,直线回归方程Y=0.513-0.002X。
结论
UTⅡ参与了家兔动脉粥样硬化形成过程,并且其血清中含逐渐降低。其与血脂水平,炎症因子IL-6血清水平呈明显负相关。在动脉粥样硬化形成过程中血脂水平明显升高,而以总胆固醇升高最为显著。炎症反应贯穿于动脉粥样硬化形成的始终,并在其中与UTⅡ相互作用,最终导致动脉粥样硬化形成。
Introduction
UrotensinⅡ(UTⅡ)was konwn to be the most vasoconstriction peptide,which widely distributed in the tissue of heart、kidney、vessel soomth muscle、even artherosclerosis plaque.It palys a key role in the mechanism of artherosclerosis plaque formation.In our study,the model of artherosclerosis model was made with rabbits,we investigate plasma change of urotensinⅡ,the relationgship between IL-6,TC.Make clear the important effect of urotensinⅡin the mechanism of artherosclerosis.
Purpose
This study designed to investigate the difference of the plasma level of urotensinⅡthe relationship between urotensinⅡ、IL-6and TC.
Material and methods
1.material
40 healty rabbits weighing 2.0-2.5kg,were obtained from department of animals,shenyang pharmaceutical university.
2.preparation of high-fat diet The basic stoyer consist 0.6%cholest,3.3% lard,8.3% egg yolk to make the high-fat diet.
3.Animal treatment
Rabbits were randomly divided into two groups,the control group and the artherosclerosis group,each group contain 20 rabbits. The control group keep feding with basic rabbit stoyer, artherosclerosis group,fedding with high-fat diet.every day,each artherosclerosis group rabbit eats 150g stoyer with contain choles t1.0g, lard 5.0g, egg yolk 12.5g.the study continue 12 weeks.
4.detection of the biochemical projects
We detected the rsbbits serum total choletrol,triglyceride,low density lipoprotein-cholesterol, high density lipoprotein-cholesterol. urotensinll and IL-6 were determined by elisa method.
5.statistical analysis
The result were processed by SPSS13.0 software.differences between the group were analyzed by Analysis of Variance(ANOVA).
Results
1.Difference of body weight and serom lipid of rabbits in each group
The difference of weight between rach has no significant sense. (P>0.05).serum lipid especially TC and LDLC had predominance higher when compare to control group and artherosclerosis grou.12weeks experimental group make obvious higher than 6weeks experimental group. (P<0.01)。
2.Difference of plasma UTⅡand IL-6 in each group
The plasma level of UTⅡin high-fat diet group goes down at 3weeks.12weeks level make big difference between control group and 6weeks group, (P< 0.01).Compared 6weeks and control group level of IL-6,the 12week high-fat diet group increased significant, (P<0.01)
3.the relationship between UTⅡ、L-6 and TC
In our study we can see negative relation between UTⅡand TC, regression equation Y=0.211-0.003X r=-0.99, P=0.01. there was also negative relation between UTⅡand IL-6 regression equation Y=0.513-0.002X,r=-0.95, P=0.04。
conclusion
UTⅡparticipate the foremation of artherosclerosis,goes decrese of plasma level. UTⅡmake obviouse negative relation between serum lipid and IL-6.There was significant upgrade of serum lipid level, especially TC.The inflammation reaction penetrate the formation of artherosclerosis,they interact tightly,make key role in artherosclerosis formation.
尾加压素Ⅱ(urotensinⅡ, UTⅡ)最早从鱼尾部下垂体中提取出的神经肽,是目前所知的体内最强的缩血管活性肽。其受体广泛分布于哺乳动物的心脏、肾脏、神经组织、血管平滑肌组织、甚至于动脉粥样硬化斑块中。UTⅡ在动脉粥样硬化中发生发展中扮演中重要角色。本实验中我们通过复制家兔动脉粥样硬化模型,观察从正常家兔直至致病过程中尾加压素动态改变,及其与血脂变化,炎症因子含量的相关性,探讨尾加压素在致动脉粥样硬化中的重要意义。
目的
研究动脉粥样硬化模型中UTⅡ血浆含量的动态变化,及其与血脂蛋白炎症因子的相关性分析。
材料方法
1.材料
健康日本大耳白家兔40只,4月龄,雌雄不限,购自沈阳市药科大学,合格证号：SCXK(辽)2009-0002,体重2.0-2.5Kg。
2.高脂饲料的配制
基础饲料中加入胆固醇(国药化学试剂有限公司,上海产,分析纯)、猪油(市售)、蛋黄粉(市售),三种成分按0.6%,3.3%,8.3%配比制成,称为高脂饮食。
3.实验动物处理
随机将家兔分为2组,对照组20只,实验组20只。单笼饲养,自由摄取饮水。实验组家兔饲喂高脂饲料,对照组继续饲喂基础兔饲料。每日每只家兔进食150g,实验组保证每日每只家兔进食胆固醇1.0g,猪油5.0g,蛋黄粉12.5g。总共喂养12周。
4.生化项目检测
于实验的0周、3周、6周、9周、12周抽血留取血浆标本,测定血清TC、HDLC、LDLC水平；应用酶联免疫法测定UTⅡ、IL-6水平。同时进行UTⅡ与血脂蛋白、炎症因子相关性分析。
5.统计学分析
应用SPSS13.0统计学软件,组间比较采用方差分析。
实验结果
1.家兔体重变化,血脂含量变化
喂养12周后家兔体重均增加,但两组间无差异(P>0.05)。实验组家兔血脂水平总体呈升高趋势。6周时,与对照组相比,实验组家兔血脂水平均升高,差别有统计学意义(P<0.05),12周实验组家兔与6周实验组家兔比较,TC和LDLC差别显著(P均<0.01)。
2.家兔血浆炎症因子UTⅡ及IL-6含量的变化
高脂饮食喂养的家兔在3周时血浆中UTⅡ含量开始下降,第12周较对照组及第6周可见其含量明显减少,有显著差异(P均<0.01)。炎症因子IL-6含量在6周时开始升高,但差别无统计学意义(P>0.05),9周及12周可见炎症因子含量明显增高,与对照组及6周实验组有显著差别(P均<0.01)
3.UTⅡ与TC、IL-6相关性分析
家兔动脉粥样硬化模型中UTⅡ与TC血清中含量呈明显负相关, r=-0.99,P=0.01,直线回归方程Y=0.211-0.003X。UTⅡ与IL-6血清中含量呈明显负相关,r=-0.95,P=0.04,直线回归方程Y=0.513-0.002X。
结论
UTⅡ参与了家兔动脉粥样硬化形成过程,并且其血清中含逐渐降低。其与血脂水平,炎症因子IL-6血清水平呈明显负相关。在动脉粥样硬化形成过程中血脂水平明显升高,而以总胆固醇升高最为显著。炎症反应贯穿于动脉粥样硬化形成的始终,并在其中与UTⅡ相互作用,最终导致动脉粥样硬化形成。
Introduction
UrotensinⅡ(UTⅡ)was konwn to be the most vasoconstriction peptide,which widely distributed in the tissue of heart、kidney、vessel soomth muscle、even artherosclerosis plaque.It palys a key role in the mechanism of artherosclerosis plaque formation.In our study,the model of artherosclerosis model was made with rabbits,we investigate plasma change of urotensinⅡ,the relationgship between IL-6,TC.Make clear the important effect of urotensinⅡin the mechanism of artherosclerosis.
Purpose
This study designed to investigate the difference of the plasma level of urotensinⅡthe relationship between urotensinⅡ、IL-6and TC.
Material and methods
1.material
40 healty rabbits weighing 2.0-2.5kg,were obtained from department of animals,shenyang pharmaceutical university.
2.preparation of high-fat diet The basic stoyer consist 0.6%cholest,3.3% lard,8.3% egg yolk to make the high-fat diet.
3.Animal treatment
Rabbits were randomly divided into two groups,the control group and the artherosclerosis group,each group contain 20 rabbits. The control group keep feding with basic rabbit stoyer, artherosclerosis group,fedding with high-fat diet.every day,each artherosclerosis group rabbit eats 150g stoyer with contain choles t1.0g, lard 5.0g, egg yolk 12.5g.the study continue 12 weeks.
4.detection of the biochemical projects
We detected the rsbbits serum total choletrol,triglyceride,low density lipoprotein-cholesterol, high density lipoprotein-cholesterol. urotensinll and IL-6 were determined by elisa method.
5.statistical analysis
The result were processed by SPSS13.0 software.differences between the group were analyzed by Analysis of Variance(ANOVA).
Results
1.Difference of body weight and serom lipid of rabbits in each group
The difference of weight between rach has no significant sense. (P>0.05).serum lipid especially TC and LDLC had predominance higher when compare to control group and artherosclerosis grou.12weeks experimental group make obvious higher than 6weeks experimental group. (P<0.01)。
2.Difference of plasma UTⅡand IL-6 in each group
The plasma level of UTⅡin high-fat diet group goes down at 3weeks.12weeks level make big difference between control group and 6weeks group, (P< 0.01).Compared 6weeks and control group level of IL-6,the 12week high-fat diet group increased significant, (P<0.01)
3.the relationship between UTⅡ、L-6 and TC
In our study we can see negative relation between UTⅡand TC, regression equation Y=0.211-0.003X r=-0.99, P=0.01. there was also negative relation between UTⅡand IL-6 regression equation Y=0.513-0.002X,r=-0.95, P=0.04。
conclusion
UTⅡparticipate the foremation of artherosclerosis,goes decrese of plasma level. UTⅡmake obviouse negative relation between serum lipid and IL-6.There was significant upgrade of serum lipid level, especially TC.The inflammation reaction penetrate the formation of artherosclerosis,they interact tightly,make key role in artherosclerosis formation.
引文
1 Ames RS,Sarau HM,Douglas SA,et al.Human urotensin Ⅱ is a potent vasoconstrictor and agonist for the orphan receptor GPR14.Naturc.1999;401:282-286.
2 Sauzeau V,Le Mellionnec E.Bertoglio J,Scalbert E.Human urotensin Ⅱ-induced contraction and arterial smooth muscle cell proliferation are mediated by RhoA and Rho-kianse.Cricle Res.2001;88;1102-1104.
3 Wanatabe T,Pakala R,Katagiri T,Benedict CR,Synerdistic effect of urotensin Ⅱ with mildly oxidized LDL on DNA synthesis in vascular smooth muscle cells.Circulation.2001;104:16-18.
4 Nicolas B,Pedro D,Robert S,Zhipeng Y.Urotensin Ⅱ receptor knockout mice on an ApoE knockout backgrand fed a high-fet diet exhibit an enhance hyperlipidemic and atherosclerotic phenotype. Circ Res.2009 Sep 25;105(7):686-95,19 p following 695. Epub 2009 Aug 20.
5 Dominique J,Thao H,Nambi A,Brian QSteven D.Urotensin Ⅱ levels in acute coronary syndromes.International J of cardiology 2006(108):31-35.
6 张丽芳,陈莉,丁文惠。血浆尾加压素Ⅱ浓度与冠状动脉病变程度的关系。Journal of China-Japan Friendship Hospital。2008 (22):3-6.
7 Hassan GS, DouglasSA。 Ohlstein EH, et al. Expression of urotensin Ⅱ in human Coronary atherosclerosis[J]. Peptid,2005(12):2464-2472.
8 Ikeda U, Ito T, Shimada K. Interleukin-6 and acute coronary sydrome. Clin Cardiol,2001, 24:701-704.
9 Bousette N。 Patel L, Douglas SA, et al.. Inereascd expression of untensin Ⅱ and its cognate receptor GPR14 in atherosclerotic lesions of the human aorta. Atheroacleresis,2004,176(1): 117-123.
10 Jones DG, AO Z,Naselsky,D,Herold CL,Maniscalco K,sarov-Blat L,Steplewski K,Airyar N,Douglas SA.Urotesin-Ⅱ-mediated cardiomyocyte hypertrophy:effect of receptor antangonism and role of inflammatory mediators.Schmiedebergs Arch pharmacol 2004;370:238-50.
11 Mallamaci F,Cutrupi S,Pizzini P,Tripepi G,Zoccali C.Urotensin Ⅱ and biomarker of endothelial activation and atherosis in endstage renal disease.Am J Hypertens 2006;19:505-10
12 Colzel M,Hess P,Qin C,et al.The urotensinllreceptor antagonist palosuran improves pancreatic and renal function in diabetes rats [J].Pharmacol Exp Ther,2006; 316 (3):1115-1121
13 Rakowaki E,Dhanak D,Hassan GS,et al. A role for urotensin Ⅱ in restenosis following balloon angioplasty:use of a selective UT receptor blocker.J Mol Cell Cardiol..2005,39(5):785-91.
1 Ames RS,Sarau HM,Douglas SA,et al.Human urotensin Ⅱ is a potent vasoconstrictor and agonist for the orphan receptor GPR14.Nature.1999;401:282-286.
2 Sauzeau V,Le Mellionnec E,Bertoglio J,Scalbert E.Human urotensin Ⅱ-induced contraction and arterial smooth muscle cell proliferation are mediated by RhoA and Rho-kianse.Cricle Res.2001;88;1102-1104.
3 Wanatabe T,Pakala R,Katagiri T,Benedict CR,Synerdistic effect of urotensin Ⅱ with mildly oxidized LDL on DNA synthesis in vascular smooth muscle cells.Circulation.2001; 104:16-18.
4 Nicolas B,Pedro D,Robert S,Zhipeng Y.Urotensin Ⅱ receptor knockout mice on an ApoE knockout backgrand fed a high-fet diet exhibit an enhance hyperlipidemic and atherosclerotic phenotype. Circ Res.2009 Sep 25; 105(7):686-95,19 p following 695. Epub 2009 Aug 20.
5 Bousette N,Patel L,Douglas SA,Ohistein EH,Giaid A.Increase expression of urotensim Ⅱ and its cognate receptor GPR14 in atherosclerotic lesion of the human aorta.Atherosclerosis 2004;176:117-23.
6 高宇,黄丽红,高长斌.尾加压素Ⅱ对大鼠血管平滑肌细胞增殖及凋亡的影响。中国老年学杂志2009年(29)：1089-91.
7 Rakowski E,Hassan GS,Dhanak D,Ohlstein EH,Douglas SA,Giaid A.A role for urotensin Ⅱ in restenosis following ballon angioplasty:use of a selective UT remceptor blocker.J Mol Cell Cardiol 2005;39;785-91.
8 Shi L,Ding W,Li D,Wang Z,Jiang H,Zhang J,Tang C.Proliferation ang anti-apoptic effect of human urotensin Ⅱ ion human endothelial cells.Atherosclerosis 2006; 188:260-4.
9 segain IP,Rolli-Derkinderen M,Gervois N,et al.Urotensin Ⅱ is a new chemotactic factor UT receptor-expressing monocytes Journal of Immunol 2007,179(2):901-909.
10温进坤,韩梅.血管平滑肌细胞[M].北京：科学出版社,2005：321-2.
11 Watanabe T, SuguroT, Kanome T,Sakamoto Y,Kodate S,Hagiwara T, et al.Human urotensin Ⅱ accelerates foam cell formation in human monocyte-derived macropgages.Hypertension 2005;46:738-44.
12 Xu S Jiang H,Yang J,Chen S.Urotensin Ⅱ induces migration of endothelial progenitor cells via activation of the RhoA/Rho kinase pathway.Tohoku J Exp Med 2009;219(4):283-8
13 Jones DG, AO Z,Naselsky,D,Herold CL,Maniscalco K,sarov-Blat L,Steplewski K,Airyar N,Douglas SA.Urotesin-Ⅱ-mediated cardiomyocyte hypertrophy:effect of receptor antangonism and role of inflammatory mediators.Schmiedebergs Arch pharmacol 2004;370:238-50.
14 Djordjevic T,BelAiba RS,Bonello S,Pfeilschifter,J,Hess J,Gorlach A.Human urotensin His a novel activator of NADPH oxidase in human pulmonary atery smooth muscle cells.Arterioscler ThromboVasc Biol 2005;25:519-25.
15 Dominique J,Thao H,Nambi A,Brian QSteven D.Urotensin Ⅱ levels in acute coronary syndromes.International J of cardiology 2006(108):31-35.
16张丽芳,陈莉,丁文惠。血浆尾加压素Ⅱ浓度与冠状动脉病变程度的关系。Journal of China-Japan Friendship Hospital。2008 (22):3-6.
17 Hassan GS, DouglasSA。Ohlstein EH, et al. Expression of urotensin Ⅱ in human Coronary atherosclerosis[J]. Peptid,2005(12):2464-2472.
18 Suguro T,Wanatabe T,BanY, et al.Increased human urotensin Ⅱ levels are correlated with carotid atheroselerosis in essential hypertension.Am J Hypertens2007,20(2):211-217.
19 Colzel M,Hess P,Qin C,et al.The urotensin Ⅱ receptor antagonist palosuran improves pancreatic and renal function in diabetes rats [J].Pharmacol Exp Ther,2006; 316 (3):1115-1121
20 Rakowaki E,Dhanak D,Hassan GS,et al. A role for urotensin Ⅱ in restenosis following balloon angioplasty:use of a selective UT receptor blocker.J. Mol Cell Cardiol.2005,39(5):785-91.
2 Sauzeau V,Le Mellionnec E.Bertoglio J,Scalbert E.Human urotensin Ⅱ-induced contraction and arterial smooth muscle cell proliferation are mediated by RhoA and Rho-kianse.Cricle Res.2001;88;1102-1104.
3 Wanatabe T,Pakala R,Katagiri T,Benedict CR,Synerdistic effect of urotensin Ⅱ with mildly oxidized LDL on DNA synthesis in vascular smooth muscle cells.Circulation.2001;104:16-18.
4 Nicolas B,Pedro D,Robert S,Zhipeng Y.Urotensin Ⅱ receptor knockout mice on an ApoE knockout backgrand fed a high-fet diet exhibit an enhance hyperlipidemic and atherosclerotic phenotype. Circ Res.2009 Sep 25;105(7):686-95,19 p following 695. Epub 2009 Aug 20.
5 Dominique J,Thao H,Nambi A,Brian QSteven D.Urotensin Ⅱ levels in acute coronary syndromes.International J of cardiology 2006(108):31-35.
6 张丽芳,陈莉,丁文惠。血浆尾加压素Ⅱ浓度与冠状动脉病变程度的关系。Journal of China-Japan Friendship Hospital。2008 (22):3-6.
7 Hassan GS, DouglasSA。 Ohlstein EH, et al. Expression of urotensin Ⅱ in human Coronary atherosclerosis[J]. Peptid,2005(12):2464-2472.
8 Ikeda U, Ito T, Shimada K. Interleukin-6 and acute coronary sydrome. Clin Cardiol,2001, 24:701-704.
9 Bousette N。 Patel L, Douglas SA, et al.. Inereascd expression of untensin Ⅱ and its cognate receptor GPR14 in atherosclerotic lesions of the human aorta. Atheroacleresis,2004,176(1): 117-123.
10 Jones DG, AO Z,Naselsky,D,Herold CL,Maniscalco K,sarov-Blat L,Steplewski K,Airyar N,Douglas SA.Urotesin-Ⅱ-mediated cardiomyocyte hypertrophy:effect of receptor antangonism and role of inflammatory mediators.Schmiedebergs Arch pharmacol 2004;370:238-50.
11 Mallamaci F,Cutrupi S,Pizzini P,Tripepi G,Zoccali C.Urotensin Ⅱ and biomarker of endothelial activation and atherosis in endstage renal disease.Am J Hypertens 2006;19:505-10
12 Colzel M,Hess P,Qin C,et al.The urotensinllreceptor antagonist palosuran improves pancreatic and renal function in diabetes rats [J].Pharmacol Exp Ther,2006; 316 (3):1115-1121
13 Rakowaki E,Dhanak D,Hassan GS,et al. A role for urotensin Ⅱ in restenosis following balloon angioplasty:use of a selective UT receptor blocker.J Mol Cell Cardiol..2005,39(5):785-91.
1 Ames RS,Sarau HM,Douglas SA,et al.Human urotensin Ⅱ is a potent vasoconstrictor and agonist for the orphan receptor GPR14.Nature.1999;401:282-286.
2 Sauzeau V,Le Mellionnec E,Bertoglio J,Scalbert E.Human urotensin Ⅱ-induced contraction and arterial smooth muscle cell proliferation are mediated by RhoA and Rho-kianse.Cricle Res.2001;88;1102-1104.
3 Wanatabe T,Pakala R,Katagiri T,Benedict CR,Synerdistic effect of urotensin Ⅱ with mildly oxidized LDL on DNA synthesis in vascular smooth muscle cells.Circulation.2001; 104:16-18.
4 Nicolas B,Pedro D,Robert S,Zhipeng Y.Urotensin Ⅱ receptor knockout mice on an ApoE knockout backgrand fed a high-fet diet exhibit an enhance hyperlipidemic and atherosclerotic phenotype. Circ Res.2009 Sep 25; 105(7):686-95,19 p following 695. Epub 2009 Aug 20.
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