活化蛋白-1在大鼠內(nèi)毒素休克急性肺損傷時(shí)血紅素氧合酶-1表達(dá)中的作用
發(fā)布時(shí)間：2018-08-08 20:24
【摘要】：內(nèi)毒素誘發(fā)的急性肺損傷／急性呼吸窘迫綜合征(ALI/ARDS)是臨床上常見(jiàn)的急危重癥,是感染性休克導(dǎo)致死亡的主要原因之一[1],因此探索其有效的防治措施具有一定的臨床意義。 血紅素氧合酶-1(HO-1)為誘導(dǎo)型,是一種應(yīng)激蛋白,又被稱(chēng)為熱休克蛋白-32(heat shock protein-32, HSP-32),其廣泛存在于各種細(xì)胞和組織。HO-1是催化血紅素降解為CO、膽紅素和鐵的限速酶,HO-1及其催化產(chǎn)物是機(jī)體重要的內(nèi)源性防御保護(hù)系統(tǒng),在維持細(xì)胞穩(wěn)定和調(diào)節(jié)炎癥反應(yīng)中發(fā)揮重要作用[2-3]。除血紅素外,內(nèi)毒素、休克、應(yīng)激、高氧等也可誘導(dǎo)HO-1表達(dá)。有研究發(fā)現(xiàn),LPS誘導(dǎo)的ALI大鼠中,α硫辛酸可通過(guò)上調(diào)HO-1的表達(dá)發(fā)揮肺組織保護(hù)作用[4]。本課題組前期研究發(fā)現(xiàn),內(nèi)毒素休克大鼠肺組織中HO-1表達(dá)上調(diào)并對(duì)肺臟發(fā)揮保護(hù)作用[5-6]。 活化蛋白-1(AP-1)是信號(hào)轉(zhuǎn)導(dǎo)通路中重要的轉(zhuǎn)錄因子復(fù)合物,能與許多基因上的AP-1位點(diǎn)結(jié)合,在細(xì)胞中發(fā)揮轉(zhuǎn)錄作用[7]。脂多糖(LPS)刺激大鼠RAW264.7巨噬細(xì)胞時(shí),活化的轉(zhuǎn)錄因子AP-1與HO-1基因啟動(dòng)子區(qū)的AP-1結(jié)合位點(diǎn)結(jié)合,可導(dǎo)致HO-1表達(dá)上調(diào)并發(fā)揮細(xì)胞保護(hù)作用[8-1ol。然而在LPS誘導(dǎo)的大鼠內(nèi)毒素休克急性肺損傷模型中,AP-1是否也參與了HO-1表達(dá)上調(diào)尚未定論。因此本研究擬在建立大鼠內(nèi)毒素休克急性肺損傷模型的基礎(chǔ)上,評(píng)價(jià)轉(zhuǎn)錄因子AP-1在HO-1表達(dá)中的作用,為探討內(nèi)毒素休克急性肺損傷時(shí)機(jī)體內(nèi)源性保護(hù)機(jī)制提供參考。 目的探討轉(zhuǎn)錄因子AP-1在大鼠內(nèi)毒素休克ALI時(shí)HO-1表達(dá)中的作用。 方法健康清潔級(jí)雄性Sprague Dawley (SD)大鼠48只,體重200-220g,2.5-3.0月齡,采用隨機(jī)數(shù)字表法,將其隨機(jī)分為4組(n=12)：正常對(duì)照組(C組)、內(nèi)毒素休克急性肺損傷組(ES組)、姜黃素+內(nèi)毒素休克急性肺損傷組(Cur+ES組)、姜黃素組(Cur組)。正常對(duì)照組(C組)腹腔注射0.1%二甲基亞砜(姜黃素溶媒)0.5ml,30min時(shí)股靜脈注射生理鹽水(LPS溶媒)0.5ml；內(nèi)毒素休克肺損傷組(ES組)腹腔注射0.1%二甲基亞砜0.5ml,30min時(shí)股靜脈注射10mg/kg LPS0.5ml;姜黃素+內(nèi)毒素休克肺損傷組(Cur+ES組)腹腔注射20mg/kg姜黃素0.5ml,30min時(shí)股靜脈注射10mg/kg LPS0.5ml;姜黃素組(Cur組)腹腔注射姜黃素20mg/kg,30min時(shí)股靜脈注射生理鹽水0.5ml。觀察動(dòng)物一般情況,持續(xù)監(jiān)測(cè)MAP和心率變化,靜脈注射LPS2h內(nèi)平均動(dòng)脈壓(MAP)較基礎(chǔ)值降低25%以下并維持該水平為休克模型制備成功標(biāo)志。本實(shí)驗(yàn)中若給予LPS后未滿(mǎn)足上述條件或6h內(nèi)死亡者均排除本觀察組。靜脈注射LPS6h時(shí)采集動(dòng)脈血0.5ml行血?dú)夥治鲇?jì)算氧合指數(shù),隨后處死大鼠留取肺組織,觀察肺組織病理學(xué)結(jié)果,行肺損傷程度評(píng)分、計(jì)算肺組織含水率、測(cè)定肺組織MDA含量和SOD活性；采用Western blot免疫印跡法分別測(cè)定大鼠肺組織HO-1和AP-1蛋白的表達(dá)；采用反轉(zhuǎn)錄酶-聚合酶鏈鎖反應(yīng)(reverse transcription-polymerase chain reaction,RT-PCR)法測(cè)定大鼠肺組織HO-1mRNA表達(dá)。 結(jié)果與C組比較,ES組和Cur+ES組肺損傷程度評(píng)分、肺組織含水率、氧合指數(shù)和MDA含量升高,SOD活性降低,AP-1、HO-1和HO-1mRNA表達(dá)上調(diào)(P0.05),Cur組上述各指標(biāo)差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)；與ES組比較,Cur+ES組肺損傷程度評(píng)分、肺組織含水率和MDA含量升高,SOD活性降低,AP-1、HO-1和HO-1mRNA表達(dá)下調(diào)(P0.05)；與Cur+ES組比較,Cur組肺損傷程度評(píng)分、肺組織含水率和MDA含量降低,氧合指數(shù)和SOD活性升高,AP-1、HO-1和HO-1mRNA表達(dá)下調(diào)(P0.05)。 結(jié)論LPS誘導(dǎo)大鼠內(nèi)毒素休克ALI時(shí),轉(zhuǎn)錄因子AP-1參與調(diào)節(jié)HO-1表達(dá)上調(diào)。
[Abstract]:Acute lung injury / acute respiratory distress syndrome (ALI/ARDS) induced by endotoxin is a common acute critical disease in clinic. It is one of the main causes of death in septic shock, [1]. Therefore, it is of certain clinical significance to explore its effective prevention and treatment measures.
Heme oxygenase -1 (HO-1) is an inducible type, which is a stress protein, also known as the heat shock protein -32 (heat shock protein-32, HSP-32). It widely exists in all kinds of cells and tissue.HO-1 to catalyze heme degradation to CO, bilirubin and iron, and HO-1 and its catalytic products are important endogenous defensive protection systems in the body. HO-1 expression can also be induced by endotoxin, shock, stress, hyperoxia and so on, which plays an important role in cell stability and regulation of inflammatory response. It is found that in LPS induced ALI rats, alpha lipoic acid can play a protective role in lung tissue by up regulation of HO-1 expression [in the early study of the group of 4]. subjects, endotoxic shock rats The expression of HO-1 in lung tissue was up-regulated and the lung was protected. [5-6].
Activated protein -1 (AP-1) is an important transcription factor complex in signal transduction pathway. It can be combined with AP-1 loci on many genes. When [7]. lipopolysaccharide (LPS) stimulates rat RAW264.7 macrophages in cells, the activated transcription factor AP-1 is combined with the AP-1 binding site in the promoter region of HO-1 gene, which can lead to the expression of HO-1. [8-1ol., however, plays an important role in LPS induced acute lung injury induced by endotoxin shock in rats, and whether AP-1 has also been involved in the up regulation of HO-1 expression. Therefore, the purpose of this study is to evaluate the role of the transcription factor AP-1 in the expression of HO-1 on the basis of the model of acute lung injury in rats with endotoxic shock. The endogenous protective mechanisms of toxin shock in acute lung injury provide reference.
Objective to investigate the role of transcription factor AP-1 in the expression of HO-1 during endotoxin shock ALI in rats.
Methods 48 healthy and clean male Sprague Dawley (SD) rats, weight 200-220g, 2.5-3.0 month old, were randomly divided into 4 groups (n=12): normal control group (C group), acute lung injury group (ES group), curcumin + endotoxin shock group (Cur+ES group), curcumin group (Cur group). Normal control group (C). Group) intraperitoneal injection of 0.1% two methyl sulfoxide (curcumin solvent) 0.5ml, 30min femoral vein injection of physiological saline (LPS solvent) 0.5ml; endotoxin shock lung injury group (ES group) intraperitoneal injection of 0.1% two methyl sulfoxide (0.5ml), 30min in the femoral vein injection 10mg/kg LPS0.5ml; curcumin + endotoxic shock lung injury group (Cur+ES group) intraperitoneal injection 20mg/kg curcumin 0.5ml, 30min, 10mg/kg LPS0.5ml were injected into the femoral vein, the curcumin group (group Cur) was injected with curcumin 20mg/kg, and the normal saline 0.5ml. was injected into the femoral vein in 30min to observe the animal general condition, and the changes of MAP and heart rate were monitored continuously. The mean arterial pressure (MAP) within the intravenous injection LPS2h was lower than the base value by 25% and maintained the level as a shock model. In this experiment, if LPS was not satisfied with the above conditions or the deaths in 6h were all excluded from the observation group, the blood gas analysis of arterial blood was collected to calculate the oxygenation index, and then the rats were sacrificed to leave the lung tissue, to observe the pathological results of the lung tissue, to score the degree of lung injury, to calculate the water content of the lung tissue and to determine the lung tissue. MDA content and SOD activity were organized and the expression of HO-1 and AP-1 protein in lung tissue of rats was measured by Western blot immunoblotting, and the lung tissue HO-1mRNA expression was measured by reverse transcriptase polymerase chain reaction (reverse transcription-polymerase chain reaction, RT-PCR).
Results compared with group C, the level of lung injury in group ES and group Cur+ES, lung tissue water content, oxygenation index and MDA content increased, SOD activity decreased, AP-1, HO-1 and HO-1mRNA expression up up (P0.05), and there was no statistical significance (P0.05) in the above-mentioned indexes of Cur group (P0.05). High SOD activity decreased, AP-1, HO-1 and HO-1mRNA were down regulated (P0.05). Compared with group Cur+ES, the degree of lung injury in Cur group, the decrease of lung tissue water content and MDA content, the increase of oxygenation index and SOD activity, AP-1, HO-1, and HO-1mRNA expression downregulation.
Conclusion transcription factor AP-1 participates in the regulation of HO-1 expression in LPS induced endotoxin shock ALI in rats.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類(lèi)號(hào)】：R459.7
本文編號(hào)：2172950
[Abstract]:Acute lung injury / acute respiratory distress syndrome (ALI/ARDS) induced by endotoxin is a common acute critical disease in clinic. It is one of the main causes of death in septic shock, [1]. Therefore, it is of certain clinical significance to explore its effective prevention and treatment measures.
Heme oxygenase -1 (HO-1) is an inducible type, which is a stress protein, also known as the heat shock protein -32 (heat shock protein-32, HSP-32). It widely exists in all kinds of cells and tissue.HO-1 to catalyze heme degradation to CO, bilirubin and iron, and HO-1 and its catalytic products are important endogenous defensive protection systems in the body. HO-1 expression can also be induced by endotoxin, shock, stress, hyperoxia and so on, which plays an important role in cell stability and regulation of inflammatory response. It is found that in LPS induced ALI rats, alpha lipoic acid can play a protective role in lung tissue by up regulation of HO-1 expression [in the early study of the group of 4]. subjects, endotoxic shock rats The expression of HO-1 in lung tissue was up-regulated and the lung was protected. [5-6].
Activated protein -1 (AP-1) is an important transcription factor complex in signal transduction pathway. It can be combined with AP-1 loci on many genes. When [7]. lipopolysaccharide (LPS) stimulates rat RAW264.7 macrophages in cells, the activated transcription factor AP-1 is combined with the AP-1 binding site in the promoter region of HO-1 gene, which can lead to the expression of HO-1. [8-1ol., however, plays an important role in LPS induced acute lung injury induced by endotoxin shock in rats, and whether AP-1 has also been involved in the up regulation of HO-1 expression. Therefore, the purpose of this study is to evaluate the role of the transcription factor AP-1 in the expression of HO-1 on the basis of the model of acute lung injury in rats with endotoxic shock. The endogenous protective mechanisms of toxin shock in acute lung injury provide reference.
Objective to investigate the role of transcription factor AP-1 in the expression of HO-1 during endotoxin shock ALI in rats.
Methods 48 healthy and clean male Sprague Dawley (SD) rats, weight 200-220g, 2.5-3.0 month old, were randomly divided into 4 groups (n=12): normal control group (C group), acute lung injury group (ES group), curcumin + endotoxin shock group (Cur+ES group), curcumin group (Cur group). Normal control group (C). Group) intraperitoneal injection of 0.1% two methyl sulfoxide (curcumin solvent) 0.5ml, 30min femoral vein injection of physiological saline (LPS solvent) 0.5ml; endotoxin shock lung injury group (ES group) intraperitoneal injection of 0.1% two methyl sulfoxide (0.5ml), 30min in the femoral vein injection 10mg/kg LPS0.5ml; curcumin + endotoxic shock lung injury group (Cur+ES group) intraperitoneal injection 20mg/kg curcumin 0.5ml, 30min, 10mg/kg LPS0.5ml were injected into the femoral vein, the curcumin group (group Cur) was injected with curcumin 20mg/kg, and the normal saline 0.5ml. was injected into the femoral vein in 30min to observe the animal general condition, and the changes of MAP and heart rate were monitored continuously. The mean arterial pressure (MAP) within the intravenous injection LPS2h was lower than the base value by 25% and maintained the level as a shock model. In this experiment, if LPS was not satisfied with the above conditions or the deaths in 6h were all excluded from the observation group, the blood gas analysis of arterial blood was collected to calculate the oxygenation index, and then the rats were sacrificed to leave the lung tissue, to observe the pathological results of the lung tissue, to score the degree of lung injury, to calculate the water content of the lung tissue and to determine the lung tissue. MDA content and SOD activity were organized and the expression of HO-1 and AP-1 protein in lung tissue of rats was measured by Western blot immunoblotting, and the lung tissue HO-1mRNA expression was measured by reverse transcriptase polymerase chain reaction (reverse transcription-polymerase chain reaction, RT-PCR).
Results compared with group C, the level of lung injury in group ES and group Cur+ES, lung tissue water content, oxygenation index and MDA content increased, SOD activity decreased, AP-1, HO-1 and HO-1mRNA expression up up (P0.05), and there was no statistical significance (P0.05) in the above-mentioned indexes of Cur group (P0.05). High SOD activity decreased, AP-1, HO-1 and HO-1mRNA were down regulated (P0.05). Compared with group Cur+ES, the degree of lung injury in Cur group, the decrease of lung tissue water content and MDA content, the increase of oxygenation index and SOD activity, AP-1, HO-1, and HO-1mRNA expression downregulation.
Conclusion transcription factor AP-1 participates in the regulation of HO-1 expression in LPS induced endotoxin shock ALI in rats.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類(lèi)號(hào)】：R459.7
【參考文獻(xiàn)】
相關(guān)期刊論文 前4條
1 季方兵,劉少華;內(nèi)毒素性急性肺損傷大鼠肺HO-1的表達(dá)及其意義[J];實(shí)用臨床醫(yī)藥雜志;2005年09期
2 黃新莉;周曉紅;周君琳;羨曉輝;丁春華;;CCK-8上調(diào)LPS誘導(dǎo)的大鼠肺組織HO-1表達(dá)的信號(hào)轉(zhuǎn)導(dǎo)機(jī)制[J];中國(guó)病理生理雜志;2009年12期
3 中華醫(yī)學(xué)會(huì)呼吸病學(xué)分會(huì),劉又寧;急性肺損傷/急性呼吸窘迫綜合征的診斷標(biāo)準(zhǔn)(草案)[J];中華結(jié)核和呼吸雜志;2000年04期
4 ;Heme oxygenase-1 in cholecystokinin-octapeptipe attenuated injury of pulmonary artery smooth muscle cells induced by lipopolysaccharide and its signal transduction mechanism[J];World Journal of Gastroenterology;2004年12期
，本文編號(hào)：2172950
本文鏈接：http://sikaile.net/yixuelunwen/jjyx/2172950.html
最近更新
教材專(zhuān)著
