中国组织工程研究与临床康复
中國組織工程研究與臨床康複
중국조직공정연구여림상강복
JOURNAL OF CLINICAL REHABILITATIVE TISSUE ENGINEERING RESEARCH
2009年
46期
9076-9079
,共4页
曾绩娟%杨博华%朱凌群%李丽娜%李玉锋%陈圆
曾績娟%楊博華%硃凌群%李麗娜%李玉鋒%陳圓
증적연%양박화%주릉군%리려나%리옥봉%진원
下肢缺血%大鼠模型%血管内皮细胞生长因子%变化规律
下肢缺血%大鼠模型%血管內皮細胞生長因子%變化規律
하지결혈%대서모형%혈관내피세포생장인자%변화규률
背景:组织缺血初期,机体通过代偿性调节与修复,实现血管的新生,使组织供血得以维持,在此代偿过程中,多种细胞因子,特别是血管内皮细胞生长因子发挥了关键的作用.目的:观察下肢缺血大鼠模型造模后不同时间点缺血组织及血清中血管内皮细胞生长因子水平变化特点及规律.设计、时间及地点:随机对照动物实验,于2007-04/11在东直门医院重点学科实验室和北京中医药大学分子生物实验室完成.材料:将4周龄雄性SD大鼠42只按照随机数字表分为6组:对照组、造模后4h、3d、1周、2周及4周组.方法:将模型组大鼠用100g/L水合氯醛麻醉后行左侧股总动脉结扎离断术制作下肢缺血模型,于造模成功后4h、3d、1周、2周及4周抽取大鼠腹主动脉血5mL,3 000r/min离心10min后,取上清;同时分离大鼠左小腿腓肠肌组织.对照组大鼠麻醉后直接取材.主要观察指标:利用Western Blotting法检测缺血组织血管内皮细胞生长因子蛋白表达量、ELASA法检测血清中血管内皮细胞生长因子水平.结果:缺血组织血管内皮细胞生长因子在缺血后4h即开始增多,3d时达到高峰,3d后血管内皮细胞生长因子表达量逐渐减少,至缺血2周时血管内皮细胞生长因子表达量最少,2周后缓慢回升,至缺血4周时血管内皮细胞生长因子表达量接近正常组织.缺血后血清血管内皮细胞生长因子水平立即下降,缺血后即刻至缺血4h下降幅度最大,缺血4h～1周下降幅度减少:缺血1～4周血清血管内皮细胞生长因子少量增加,至缺血4周时仍显著低于正常组(P＜0.01).血清血管内皮细胞生长因子含量变化趋势表现为至缺血4h迅速下降,缺血1周降至最低点,1周后缓慢升高的趋势.结论:大鼠后肢缺血后,缺血组织中血管内皮细胞生长因子水平呈现先升高-再下降-再上升的趋势,血清中血管内皮细胞生长因子水平呈现先下降-再上升趋势,即呈现血清水平低,局部组织水平高的特点.
揹景:組織缺血初期,機體通過代償性調節與脩複,實現血管的新生,使組織供血得以維持,在此代償過程中,多種細胞因子,特彆是血管內皮細胞生長因子髮揮瞭關鍵的作用.目的:觀察下肢缺血大鼠模型造模後不同時間點缺血組織及血清中血管內皮細胞生長因子水平變化特點及規律.設計、時間及地點:隨機對照動物實驗,于2007-04/11在東直門醫院重點學科實驗室和北京中醫藥大學分子生物實驗室完成.材料:將4週齡雄性SD大鼠42隻按照隨機數字錶分為6組:對照組、造模後4h、3d、1週、2週及4週組.方法:將模型組大鼠用100g/L水閤氯醛痳醉後行左側股總動脈結扎離斷術製作下肢缺血模型,于造模成功後4h、3d、1週、2週及4週抽取大鼠腹主動脈血5mL,3 000r/min離心10min後,取上清;同時分離大鼠左小腿腓腸肌組織.對照組大鼠痳醉後直接取材.主要觀察指標:利用Western Blotting法檢測缺血組織血管內皮細胞生長因子蛋白錶達量、ELASA法檢測血清中血管內皮細胞生長因子水平.結果:缺血組織血管內皮細胞生長因子在缺血後4h即開始增多,3d時達到高峰,3d後血管內皮細胞生長因子錶達量逐漸減少,至缺血2週時血管內皮細胞生長因子錶達量最少,2週後緩慢迴升,至缺血4週時血管內皮細胞生長因子錶達量接近正常組織.缺血後血清血管內皮細胞生長因子水平立即下降,缺血後即刻至缺血4h下降幅度最大,缺血4h～1週下降幅度減少:缺血1～4週血清血管內皮細胞生長因子少量增加,至缺血4週時仍顯著低于正常組(P＜0.01).血清血管內皮細胞生長因子含量變化趨勢錶現為至缺血4h迅速下降,缺血1週降至最低點,1週後緩慢升高的趨勢.結論:大鼠後肢缺血後,缺血組織中血管內皮細胞生長因子水平呈現先升高-再下降-再上升的趨勢,血清中血管內皮細胞生長因子水平呈現先下降-再上升趨勢,即呈現血清水平低,跼部組織水平高的特點.
배경:조직결혈초기,궤체통과대상성조절여수복,실현혈관적신생,사조직공혈득이유지,재차대상과정중,다충세포인자,특별시혈관내피세포생장인자발휘료관건적작용.목적:관찰하지결혈대서모형조모후불동시간점결혈조직급혈청중혈관내피세포생장인자수평변화특점급규률.설계、시간급지점:수궤대조동물실험,우2007-04/11재동직문의원중점학과실험실화북경중의약대학분자생물실험실완성.재료:장4주령웅성SD대서42지안조수궤수자표분위6조:대조조、조모후4h、3d、1주、2주급4주조.방법:장모형조대서용100g/L수합록철마취후행좌측고총동맥결찰리단술제작하지결혈모형,우조모성공후4h、3d、1주、2주급4주추취대서복주동맥혈5mL,3 000r/min리심10min후,취상청;동시분리대서좌소퇴비장기조직.대조조대서마취후직접취재.주요관찰지표:이용Western Blotting법검측결혈조직혈관내피세포생장인자단백표체량、ELASA법검측혈청중혈관내피세포생장인자수평.결과:결혈조직혈관내피세포생장인자재결혈후4h즉개시증다,3d시체도고봉,3d후혈관내피세포생장인자표체량축점감소,지결혈2주시혈관내피세포생장인자표체량최소,2주후완만회승,지결혈4주시혈관내피세포생장인자표체량접근정상조직.결혈후혈청혈관내피세포생장인자수평립즉하강,결혈후즉각지결혈4h하강폭도최대,결혈4h～1주하강폭도감소:결혈1～4주혈청혈관내피세포생장인자소량증가,지결혈4주시잉현저저우정상조(P＜0.01).혈청혈관내피세포생장인자함량변화추세표현위지결혈4h신속하강,결혈1주강지최저점,1주후완만승고적추세.결론:대서후지결혈후,결혈조직중혈관내피세포생장인자수평정현선승고-재하강-재상승적추세,혈청중혈관내피세포생장인자수평정현선하강-재상승추세,즉정현혈청수평저,국부조직수평고적특점.
BACKGROUND:In the initial stage of ischemia,organisms complete angiogenesis and maintain the blood supply of organization through compensatory regulation and repair,in which process a variety of cytokines,especially vascular endothelial growth factor (VEGF),have played a key role.OBJECTIVE:To observe the characteristics and rules of VEGF level changes in both ischemic tissues and blood serums at different time points after establishing rat models of lower limb ischemia.DESIGN,TIME AND SETTING:A randomized controlled animal experiment was performed at the Loboratory for Key Subjects in Dongzhimen Hospital and the Laboratory of Molecular Biology in Beijing University of Chinese Medicine from April to November in 2007.MATERIALS:A total of 42 male SD rats of 4 weeks were divided by random digits table into 6 groups,namely a control group and a model group which was subdivided into 5 groups at the time points of hour 4,day 3,weeks 1,2 and 4 post modeling respectivly.METHODS:Rat models of lower limb ischemia were established in the model group by performing ligation and mutilation operation to left femoral arteries of rats that were anesthetized with 100 g/L chloral hydrate. At the time points of hour 4,day 3,weeks 1,2 and 4 following modeling respectively,rats were selected to extract their abdominal aorta blood samples whose supernatant was then obtained through 10 minutes of 3 000r/min centrifugalization. Gastrocnemius tissues in left legs of rats were isolated at the same time. Samples in control group were obtained directly from anesthetized rats.MAIN OUTCOME MEASURES:Western blotting method was used for detecting protein expression of VEGF in ischemic tissues and ELASA method for VEGF level in serum.RESULTS:The protein expression of VEGF in ischemic tissues began to increase immediately at hour 4 following ischemia and reached a peak at day 3,after which it reduced gradually till week 2 when it reached its minimum. Then it began to increase again and reached the level close to that of normal tissues by the end of week 4 following ischemia. As for the VEGF level in serum,it decreased immediately after ischemia,with the maximum decrease amplitude between immediate and hour 4 following ischemia,a smaller one between hour 4 and week 1;From week 1 following ischemia on,it began to increase but was still lower than the normal level by the end of week 4 (P＜0.01 ).The VEGF level in serum changed with the tendency of immediate decrease from hour 4,minimum at week 1 and graduate increase after week 1 following ischemia.CONCLUSION:After ischemia in lower limb,VEGF level in ischemic tissues changes in the direction of increase-decrease-increase;VEGF level in serum changed in the direction of decrease-increase. In another words,VEGF levels after ischemia shows the characteristics of being low in serum and high in local ischemic tissues.
