本文選題：Apelin-13 + 正性肌力��； 參考：《寧夏醫(yī)科大學》2010年碩士論文

【摘要】：Apelin是Tatemoto等在1998年利用反向藥理學方法從牛胃分泌物中提取出來的血管活性多肽,是G蛋白偶聯(lián)受體APJ受體(putative receptor protein related to the angiotensinII receptor AT1)的內源性配體。Apelin基因位于X染色體的q25-26.1,包含3個外顯子和2個內含子。Apelin的前體蛋白由77個氨基酸組成,可水解生成不同長度的活性肽段,如apelin-36、apelin-31、apelin-17、apelin-13等。在體內,Apelin及其受體APJ廣泛分布在中樞與外周組織,如心臟、肺臟、腎臟、中樞神經(jīng)系統(tǒng)、脂肪組織等,具有調節(jié)心血管系統(tǒng)穩(wěn)態(tài)、水鹽平衡、調節(jié)免疫反應等作用,是一種重要的生理調節(jié)肽。其中在心血管組織中, apelin具有增強心肌收縮力、舒張血管、降低血壓等作用,提示該肽可能是一種重要的心血管調節(jié)肽。本工作以單個大鼠心室肌細胞為模型,應用全細胞膜片鉗技術,觀察apelin-13對大鼠心室肌細胞鈉鈣交換電流(I_(Na/Ca))、L型鈣通道電流(I_(Ca,L))、ATP敏感鉀通道電流(IK_(ATP))的影響,以進一步明確apelin對心臟正性肌力作用的離子電流機制。 目的本實驗旨在研究apelin-13對急性分離大鼠心室肌細胞正性肌力作用的離子通道機制。 方法雄性SD大鼠(250-300 g),腹腔麻醉后斷頭處死,迅速取其心臟,以膠原酶溶液灌流心臟,待心臟基本變軟后剪取心室部分,剪碎吹打使之分離成為單個心室肌細胞。使用全細胞膜片鉗技術記錄心室肌細胞的I_(Na/Ca)、I_(Ca,L)、IK_(ATP)。給藥組以含不同濃度apelin-13的細胞外液灌流心室肌細胞5 min;對照組直接以細胞外液灌流5 min,記錄并觀察電流曲線的變化。 結果 1.Apelin-13對大鼠心室肌細胞I_(Na/Ca)的影響Apelin-13對內向、外向I_(Na/Ca)均有增大作用。濃度為0.01 nmol/L、0.1 nmol/L、1 nmol/L的apelin-13使正向INaCa分別比給藥前增大20±4%( P 0.05,n=8)、33±7%( P 0.01,n=8)、54±10%( P 0.01,n=8),使反向I_(Na/Ca)分別增大1±6%(P 0.05 ,n=8),7±2%(P 0.01,n=8),30±7%(P 0.01,n=8),對照組單純給予細胞外液5 min后正、反向I_(Na/Ca)未分別減小5±4%和2±7%。 2.Apelin-13對大鼠心肌細胞I_(Ca,L)的影響Apelin-13可濃度依賴性增大大鼠心肌細胞I_(Ca,L),以apelin-13濃度分別為0.01 nmol/L、0.1 nmol/L、1 nmol/L的細胞外液灌流心肌細胞5 min后I_(Ca,L)分別增大3士3%、20士6%、45士9%,對照組由于rundown現(xiàn)象減小5士5%,0.1 nmol/L組、1 nmol/L組與對照組相比較有差異顯著(P 0.01,n=8),0.01 nmol/L與對照組無顯著差異(P 0.05,n=8)。1 nmol/L的apelin-13灌流5 min后IV曲線明顯下移,但不改變I_(Ca,L)的電壓依賴關系和反轉電位。對照I_(Ca,L)激活曲線半激活電位是-19.43±2.76mV,斜率是4.76±0.33 mV,1 nmol/L apelin-13灌流5 min后半激活電位變?yōu)?18.93±3.05 mV,斜率變?yōu)?.21±0.38 mV,二者無明顯區(qū)別(P 0.05, n=6)。對照I_(Ca,L)失活曲線半失活電位是-23.62±1.79mV,斜率是5.35±0.21 mV,1 nmol/L apelin-13灌流5 min后半失活電位變?yōu)?24.6±2.14 mV,斜率變?yōu)?.47±0.23,二者無明顯區(qū)別(P 0.05, n=6)。上述結果表明apelin-13增大I_(Ca,L)的作用不是通過改變L型鈣通道激活和失活的門控特點。 3.Apelin-13對IK_(ATP)的影響在由50 mV到-100 mV持續(xù)125 ms的斜坡刺激下記錄心肌細胞IK_(ATP)。1 nmol/L、10 nmol/L的apelin-13不能使該電流增加,給予pinacidil后該電流明顯增加,并且能被glibenelmaide阻斷(說明該電流是IK_(ATP)),提示apelin-13不能使靜息狀態(tài)下心肌細胞的ATP敏感鉀通道開放。 結論本工作以單個大鼠心室肌細胞為模型,應用全細胞膜片鉗技術,經(jīng)研究發(fā)現(xiàn),apelin-13可濃度依賴性地增大心室肌細胞的I_(Na/Ca)和I_(Ca,L),對ATP敏感鉀通道無開放作用。上述結果表明,apelin-13對心臟正性肌力作用的離子通道電流機制可能與通過增大心肌細胞I_(Na/Ca)和I_(Ca,L)電流,從而增加心肌細胞的鈣離子內流有關。
[Abstract]:Apelin is a vasoactive polypeptide extracted from bovine gastric secretions in 1998 by Tatemoto and so on. It is the endogenous ligand of the G protein coupled receptor APJ receptor (putative receptor protein related to the angiotensinII receptor), which contains 3 exons and 2 The precursor protein of intron.Apelin consists of 77 amino acids that can be hydrolyzed to produce active peptide segments of different lengths, such as apelin-36, apelin-31, apelin-17, apelin-13, etc. in vivo, Apelin and its receptor APJ are widely distributed in central and peripheral tissues, such as the heart, lungs, kidneys, central nervous system, adipose tissue, and so on, which regulate the cardiovascular system. The effect of homeostasis, water and salt balance and regulating immune response, is an important physiological regulation peptide. In cardiovascular tissue, Apelin has the effect of enhancing myocardial contractile force, diastolic blood vessel, lowering blood pressure and so on. It suggests that the peptide may be an important cardiovascular regulation peptide. Patch clamp technique was used to observe the effect of apelin-13 on sodium calcium exchange current (I_ (Na/Ca)), L type calcium channel current (I_ (Ca, L)), ATP sensitive potassium channel current (IK_ (ATP)) in ventricular myocytes of rats, in order to further clarify the ionic current mechanism of apelin on the positive inotropic action of the heart.
Objective to study the ion channel mechanism of apelin-13 on the positive inotropic effect of acute isolated ventricular myocytes in rats.
Methods the male SD rats (250-300 g) were killed after the abdominal anesthesia, and the heart was taken quickly. The heart was perfused with collagenase solution. After the heart became soft, the ventricular part was cut to separate into single ventricular myocytes. The whole cell patch clamp technique was used to record the I_ (Na/Ca) of ventricular myocytes, I_ (Ca, L), and IK_ (ATP). The extracellular fluid of different concentrations of apelin-13 Perfused Ventricular Myocytes 5 min; the control group was perfused directly with extracellular fluid for 5 min, recording and observing the changes of current curve.
Result
The effect of 1.Apelin-13 on I_ (Na/Ca) in ventricular myocytes of rats increased Apelin-13 to introverted and extroverted I_ (Na/Ca). The concentration of 0.01 nmol/L, 0.1 nmol/L, and 1 nmol/L apelin-13 made the positive INaCa 20 + 4% (P 0.05, 0.01, 54), 54 + 10% (54 + 0.01,), respectively. (n=8), 7 + 2% (P 0.01, n=8), 30 + 7% (P 0.01, n=8), the control group only given extracellular fluid 5 min, the reverse I_ (Na/Ca) did not decrease 5 5 4% and 2 2 7%. respectively.
The effect of 2.Apelin-13 on rat cardiac myocyte I_ (Ca, L), Apelin-13 could increase the concentration dependent I_ (Ca, L) of rat cardiac myocytes, and the concentration of apelin-13 was 0.01 nmol/L, 0.1 nmol/L, and 1 nmol/L was 3%, 20 6%, 45 9%, respectively. Group l/L, the 1 nmol/L group was significantly different from the control group (P 0.01, n=8), and there was no significant difference between the 0.01 nmol/L and the control group (P 0.05, n=8).1 nmol/L apelin-13 perfusion 5 min after the IV curve obviously moved down, but did not change the voltage dependence and reverse potential. .76 + 0.33 mV, 1 nmol/L apelin-13 perfusion 5 min half activation potential changed to -18.93 + 3.05 mV, the slope changed to 5.21 + 0.38 mV, and there was no obvious difference between two (P 0.05, n=6). The changes were 5.47 + 0.23, and there was no obvious difference between the two (P 0.05, n=6). The results showed that the effect of apelin-13 increasing I_ (Ca, L) was not by changing the activation and inactivation of the L type calcium channel.
The effect of 3.Apelin-13 on IK_ (ATP) was recorded from 50 mV to -100 mV sustained by the slope of 125 ms, and IK_ (ATP).1 nmol/L was recorded. The current of 10 nmol/L could not increase the current. The ATP sensitive potassium channel of cardiac myocytes is open.
Conclusion a single rat ventricular myocyte was used as a model, and the whole cell patch clamp technique was applied. It was found that apelin-13 could increase the concentration of I_ (Na/Ca) and I_ (Ca, L) in ventricular myocytes in a concentration dependent manner, and had no open effect on ATP sensitive potassium channels. The results showed that the ion channel current mechanism of apelin-13 for positive inotropic action of the heart was possible. It can be related to increasing the calcium influx of cardiac myocytes by increasing the I_ (Na/Ca) and I_ (Ca, L) currents.
【學位授予單位】：寧夏醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R33

【共引文獻】

相關期刊論文 前8條

1 朱波;王玉環(huán);呂杰強;;探討apelin與妊娠期高血壓疾病的關系[J];國外醫(yī)學(計劃生育/生殖健康分冊);2007年06期

2 ;Apelin and vascular endothelial growth factor are associated with mobilization of endothelial progenitor cells after acute myocardial infarction[J];Journal of Biomedical Research;2012年06期

3 潘菊英;彭菊蘭;劉媛媛;;妊娠期高血壓疾病患者血清Apelin水平變化及臨床意義[J];海南醫(yī)學;2013年20期

4 安松濤;李凌;齊艷艷;馬旭;;腺病毒介導的Apelin基因過表達對心力衰竭大鼠的保護作用[J];實用醫(yī)學雜志;2010年17期

5 江燕萍;孟錦繡;游可理;林燕燕;;子癇前期胎盤組織中Apelin/APJ的表達及其臨床意義[J];實用婦產(chǎn)科雜志;2013年01期

6 朱軍;王連生;;Apelin/APJ系統(tǒng)與冠心病研究進展[J];心血管病學進展;2010年05期

7 李薇薇;張怡;高平進;朱鼎良;;AGTRL1基因變異與原發(fā)性高血壓[J];中國分子心臟病學雜志;2008年05期

8 李薇薇;張怡;高平進;朱鼎良;;血清心血管活性多肽Apelin與原發(fā)性高血壓伴代謝異常的關系[J];中華高血壓雜志;2008年12期

相關博士學位論文 前5條

1 徐承啟;中國人群動脈粥樣硬化相關疾病的分子遺傳學研究[D];華中科技大學;2010年

2 張志;Apelin在急性心肌缺血損傷時的變化及其心肌保護作用機制[D];中國醫(yī)科大學;2009年

3 王孝東;Apelin對心肌缺血大鼠心臟側枝循環(huán)的影響和機制[D];中國醫(yī)科大學;2010年

4 呂心瑞;Am80調節(jié)apelin基因表達的分子機制研究[D];河北醫(yī)科大學;2013年

5 秦迪;重組AAV介導多基因促進成熟性血管新生在小鼠后肢缺血微循環(huán)中的研究[D];吉林大學;2013年

相關碩士學位論文 前10條

1 周湘鴻;NT-proBNP和Apelin-12在冠心病患者血漿中的變化及意義[D];鄭州大學;2010年

2 楊艷杰;側腦室注射apelin-13抑制小鼠遠端結腸運動[D];蘭州大學;2011年

3 趙乾;外源性Apelin-13對心力衰竭大鼠血漿血管緊張素Ⅱ和腎上腺髓質素及心功能的影響[D];蘭州大學;2011年

4 張勝葉;外源性Apelin-13對心力衰竭大鼠心功能和血漿BNP、NO的影響[D];蘭州大學;2011年

5 朱軍;1、PRKCH基因多態(tài)性與中國冠心病人群遺傳易感相關性研究 2、MCP-1/CCR2/RANTES/CCR5基因多態(tài)性與冠心病關聯(lián)性的Meta分析[D];南京醫(yī)科大學;2011年

6 羅麗萍;高血壓患者血漿Apelin-12水平與頸動脈內中膜厚度的相關性研究[D];石河子大學;2010年

7 趙蓉;香菇調脂膠囊對大鼠高脂血癥的預防作用[D];華中科技大學;2009年

8 封芬;Apelin/APJ參與大鼠心肌肥厚形成的caveolin-1-Akt信號通路研究[D];南華大學;2007年

9 劉燕;兔急性肺動脈栓塞時Apelin-APJ系統(tǒng)的變化及意義[D];河北醫(yī)科大學;2010年

10 任曉倩;Apelin-13對葡萄糖剝奪誘導的乳鼠心肌細胞自體吞噬的影響[D];遼寧醫(yī)學院;2012年

，

本文編號：1967448