血红素加氧酶-1诱导对鼠肝缺血再灌注损伤的保护作用

目的研究血红素加氧酶-1(heme oxygenase-1,HO-1)在鼠肝缺血再灌注损伤肝组织中的表达及其作用。方法建立小鼠部分肝脏热缺血再灌注损伤模型,36只清洁级Balb/C小鼠随机分为3组: 假手术组(S组)、缺血/再灌注损伤组(I/R组)、HO-1诱导剂氯化高铁血红素(hemin)预处理组(HM组)。免疫组化半定量分析肝组织HO-1蛋白的表达,检测血清AST和ALT,肝组织丙二醛(MDA)含量及超氧化物歧化酶(SOD)活性,并观察肝组织的病理变化。结果与S组比较,I/R组HO-1蛋白表达显著增强,hemin预处理后,HO-1蛋白表达较I/R组增高(P<0．01)。I/R组AST,ALT活性和MDA的含量显著高于S组,而HM组均显著低于I/R组(P<0．01);I/R组SOD活性下降,而HM组显著高于I/R组(P<0．01)。HM组病理损伤程度明显轻于I/R组。结论 HO-1在鼠肝缺血再灌注损伤肝组织中表达上调,对肝脏具有保护效应。     

缺血再灌注肾损伤中血红素加氧酶-1的表达及其意义

肾脏缺血再灌注(IR)损伤与再灌注后氧应激反应的介导有关,而肾脏IR损伤的可逆性可能与肾脏内在的抗氧化保护机制相关犤1犦。血红素加氧酶-1(hemeoxygenase-1,HO-1)在肾脏的诱导性表达可能参与这一保护机制。本研究通过建立大鼠IR损伤模型,动态观察了IR大鼠肾脏HO-1在蛋白水平的时相表达,分析HO-1在肾脏缺血再灌注可逆性损伤中的作用;并通过lazaroid的干预,初步了解氧自由基清除剂对HO-1表达的影响。一、材料与方法1.模型建立与取样:SD雄鼠60只,先采血3ml,随机分为:(1)缺血再灌注组(IR组,n=20):切除右肾,夹闭左肾动脉60min后开启;(2)…     

缺血后处理对大鼠缺血再灌注损伤肝脏血红素加氧酶-1表达的影响

灌注损伤,其机制可能与增加HO-1的表达和增强移植肝脏抗氧化能力有关.     

血红素加氧酶-1在治疗性高碳酸血症减轻大鼠肝缺血-再灌注损伤中的作用

目的探究血红素加氧酶-1(HO-1)在治疗性高碳酸血症减轻大鼠肝缺血-再灌注损伤中的作用。方法健康成年SD大鼠60只,随机均分为四组:假手术组(C组)、肝缺血-再灌注损伤组(IR组)、治疗性高碳酸血症处理组(A组)、治疗性高碳酸血症处理+锌原卟啉(ZnPP)预处理组(B组)。通过夹闭大鼠肝门静脉,肝动脉及胆管建立部分肝脏缺血-再灌注损伤模型,实验共缺血1h,再灌注6h。C组和IR组:实验全程机械通气吸入50%O2-50%N2;A组和B组:缺血即刻至再灌注结束期间吸入外源性50%O2-45%N2-5%CO2,C组、IR组、A组于实验前24h腹腔注射生理盐水5ml/kg,B组注射ZnPP 5mg/kg。于机械通气前(基础值)、缺血前即刻、再灌注即刻、再灌注后1、2、3、4、5、6h检测动脉血PaCO2;再灌注6h后取血清检测ALT、AST和TNF-α,并取肝组织HE染色观察病理学改变,TUNEL法检测细胞凋亡水平,Western blot法检测HO-1的相对量表达。结果与C组和IR组比较,A组和B组动脉血PaCO2水平明显升高(P0.05)。与C组比较,IR组肝组织损伤严重,IR组、A组和B组血清ALT、AST和TNF-α含量、凋亡指数和A组HO-1相对表达量明显升高(P0.05)。与IR组比较,A组肝组织损伤减轻,A组和B组血清ALT、AST、TNF-α含量、凋亡指数和B组的HO-1相对表达量明显降低(P0.05),而A组HO-1相对表达量明显升高(P0.05)。与A组比较,B组肝组织损伤加重,血清ALT、AST、TNF-α含量、凋亡指数明显升高(P0.05),HO-1相对表达量明显下降(P0.05)。结论治疗性高碳酸血症通过上调HO-1有效减轻肝缺血-再灌注损伤。     

小剂量氯胺酮预处理对大鼠肠缺血/再灌注后肝脏血红素加氧酶-1表达的影响

目的 观察小剂量氯胺酮对血红素加氧酶-1 (heme-oxygenase 1,HO-1)在大鼠肠缺血/再灌注(ischemia/reperfusion,I/R)后肝脏中表达的影响,并对其可能的保护机制作一初步探讨. 方法 48只雄性成年SD大鼠按随机数字表法分为A组(氯胺酮10 mg/kg术前30 min腹腔注射+假手术)、B组(盐水0.2 ml术前30 min腹腔注射+假手术)、C组(氯胺酮10 mg/kg术前30 min腹腔注射+小肠I/R)、D组(盐水0.2 ml术前30 min腹腔注射+小肠I/R)、E组(锌原卟啉5 mg/kg、氯胺酮10 mg/kg术前30 min腹腔注射+小肠I/R)、F组(锌原卟啉5 mg/kg、盐水0.2 ml术前30 min腹腔注射+小肠I/R),每组8只.小肠I/R造模组大鼠通过夹闭肠系膜上动脉60 min,然后松血管夹,于再灌注6h后取材,测定肝组织丙二醛(malondialdehyde,MDA)含量、超氧化物歧化酶(superoxide dismutase,SOD)活性;采用免疫组化法测定肝组织HO-1的表达和定量;光镜下观察肝脏病理学改变;取外周静脉血测血清谷丙转氨酶(glutamic-pyruvic transaminase,GPT)、谷草转氨酶(glutamic-oxaloacetic transaminase,GOT). 结果 A、B两组间各项数据比较差异无统计学意义(P＞0.05);各缺血造模组大鼠肝组织MDA含量:C组[(2.69±0.35) μmol/g]、D组[(4.62±0.36) μmol/g]、E组[(4.35±0.24) μmol/g]、F组[(4.74±0.34) μmol/g]与B组[(1.08±0.26) μmol/g]比较,显著升高;SOD活力:C组[(129±6) μU/L]、D组[(102±5) μU/L]、E组[(99±4)μU/L]、F组[(96±6)μU/L]与B组[(133±7) μU/L]比较,显著下降;血清GOT、GPT含量:C组[GPT(212±21) U/L,GOT(129±16) U/L]、D组[GPT(416±33) U/L,GOT(362±15)U/L]、E组[GPT(407±29) U/L,GOT(359±14) U/L]、F组[GPT(414-±40) U/L,GOT(366±16) U/L]与B组[(GPT(53±9) U/L,GOT(83±7) U/L]比较,显著升高;HO-1表达:C组(0.472±0.126)、D组(0.324±0.078)、E组(0.337±0.092)、F组(0.328±0.083) OD与B组(0.078±0.010) OD比较,显著上调(P＜0.05或P＜0.01);与D组比较,C组血清GPT、GOT、肝组织MDA含量均显著降低,SOD活力显著上升,HO-1表达上调(P＜0.05);与D组比较,E组、F组血清GPT、GOT及MDA、SOD值差异无统计学意义(D0.05);E组和C组比较,血清GPT、GOT,肝组织MDA含量显著升高,SOD活力显著下降,HO-1表达下调(P＜0.05). 结论 小剂量氯胺酮预处理能减轻肠I/R后造成的肝脏损伤,这种作用在一定程度上是通过上调HO-1的表达来实现的.     

细胞穿透肽PEP-1介导血红素加氧酶-1对大鼠肠缺血再灌注损伤的影响

目的 评价细胞穿透肽PEP-1介导血红素加氧酶-1(HO-1)对大鼠肠缺血再灌注损伤的影响.方法 雄性SD大鼠18只,周龄7～9周,体重210～260 g,采用随机数字表法,将大鼠随机分为3组(n=6):假手术组(S组)、肠缺血再灌注组(IR组)和融合蛋白PEP-1/HO-1+肠缺血再灌注组(HO组).采用夹闭肠系膜上动脉45 min,恢复灌注120 min的方法制备大鼠肠缺血再灌注损伤模型.HO组夹闭肠系膜上动脉前30 min,左侧髂静脉注射融合蛋白PEP-1/HO-1 0.5 mg,S组不夹闭肠系膜上动脉,余操作同IR组.于再灌注120 min时处死大鼠取小肠组织,称重后计算肠湿/干重比,测定丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性和HO-1活性,免疫组化法检测肠组织HO-1蛋白的表达,光镜下观察肠组织结构并进行损伤评分.结果 与S组比较,IR组和HO组肠湿/干重比和MDA含量升高,SOD活性降低,HO-1活性和蛋白表达水平升高,损伤评分升高(P＜0.05);与IR组比较,HO组肠湿/干重比、MDA含量降低,SOD活性升高,HO-1活性和蛋白表达水平升高,损伤评分降低(P＜0.05).HO组大鼠肠组织病理学损伤较IR组减轻.结论 细胞穿透肽PEP-1可将HO-1成功导人大鼠肠组织中的细胞并减轻肠缺血再灌注损伤.

Abstract：

Objective To investigate the effects of heme oxygenase-1 (HO-1) mediated by cell penetrating peptide PEP-1 on intestinal ischemia/reperfusion (I/R) injuiy in tats. Methods Eighteen male SD rats aged 7-9 weeks weighing 210-260 g were randomly divided into 3 groups (re = 6 each): sham operation group (group S) , I/R group and PEP-1/HO-1 + I/R group (group HO) . To establish a model of intestinal I/R injury, intestines were exteriorized and the superior mesenteric artery was exposed and occluded for 45 min ischemia, and then the clamp was removed for 120 min reperfusion. The PEP-1/HO-1 fusion protein 0.5 mg was injected via the left iliac vein 30 min prior to ischemia in group HO. The superior mesenteric artery was exposed but not occluded in group S. At the end of reperfusion, the rats were sacrificed and intestinal tissues obtained to determine the intestinal wet/ dry ratio, malondialdehyde (MDA) level, activities of superoxide dismutase (SOD) and HO-1, and HO-1 protein expression. The histological changes in the intestinal mucosa were examined and the injuiy was scored. Results Compared with group S, the intestinal wet/dry ratio, MDA level, HO-1 activity, HO-1 protein expression and injury score were significantly increased, while the SOD activity was significantly decreased in groups I/R and HO ( P ＜ 0.05) . Compared with group I/R, the intestinal wet/dry ratio, MDA level and injury score were significantly decreased, while the SOD activity, HO-1 activity and HO-1 protein expression increased in group HO ( P ＜ 0.05) . The pathologic changes were significantly attenuated in group HO compared with group I/R.Conclusion HO-1 protein can be successfully delivered into intestinal tissues by PEP-1 and has protective effects against intestinal I/R injury.     

辛伐他汀预处理对肢体缺血再灌注诱发肺损伤大鼠肺组织血红素加氧酶-1表达的影响

目的 评价辛伐他汀预处理对肢体缺血再灌注诱发肺损伤大鼠肺组织血红素加氧酶-1(HO-1)表达的影响.方法 成年雄性SD大鼠48只,体重250～300 g,采用随机数字表法,将大鼠随机分为6组(n=8):假手术组(S组)、肢体缺血再灌注组(IR组)、辛伐他汀1、5、10 mg/kg组(S1组、S2组、S3组)和辛伐他汀对照组(SC组).采用夹闭股动脉2 h,再灌注3 h的方法制备肢体缺血再灌注模型.S组:仅分离股动脉和股静脉,不夹闭;IR组:制备肢体缺血再灌注模型;S1组、S2组、S3组:分别将辛伐他汀1、5、10 mg/kg溶于1 ml蒸馏水,于每13清晨灌胃1次,连续灌胃3 d后制备肢体缺血再灌注模型;SC组:辛伐他汀10 mg/kg溶于1 ml蒸馏水,于每日清晨灌胃1次,连续灌胃3 d.再灌注3 h时取颈动脉血样,行血气分析,记录PaO2和PaCO2,随后处死大鼠,取肺组织,观察病理学结果,计算湿重/干重比(W/D比),测定SOD活性,计数PMN,测定HO-1 mRNA及其蛋白的表达水平.结果 与S组比较,IR组PaO2及PaCO2降低,IR组、S1组和S2组肺组织W/D比和PMN计数升高,SOD活性降低(P＜0.05),S3组和SC组上述指标差异无统计学意义(P＞0.05),IR组、S1组、S2组、S3组和SC 组肺组织H0-1 mRNA及其蛋白表达上调(P＜0.01);与IR组比较,S1组、S2组和S3组PaO2、PaCO2及肺组织SOD活性升高,肺组织W/D比和PMN计数降低,肺组织HO-1 mRNA及其蛋白表达上调(P＜0.05或0.01);S1组、S2组和S3组肺组织WID比和PMN计数依次降低,SOD活性依次升高,HO-1 mRNA及其蛋白表达依次上调(P＜0.05或0.01).S1组、S2组和S3组肺组织病理性损伤较IR组减轻.结论 辛伐他汀预处理可上调肢体缺血再灌注大鼠肺组织HO-1的表达,从而产生肺保护作用,且呈剂量依赖性.

Abstract：

Objective To investigate the effects of simvastatin preconditioning on the pulmonary heme oxygenase-1 (HO-1) expression in rats with lung injury induced by ischemia-reperfusion (I/R) of hind limbs. Methods Forty-eight adult male SD rats weighing 250-300 g were randomly divided into 6 groups ( n = 8 each) : sham operation group (group S) ; I/R group; I/R + simvastatin 1,5, 10 mg/kg groups (S1 , S2, S3 groups) ; simvastatin control group (group SC) . I/R of hind limbs was produced by occlusion of bilateral femoral arteries for 2 h followed by 3 h reperfusion. Croups S1 , S2 , S3 received simvastatin 1, 5, 10 mg/kg respectively via an oro-gastric tube for 3 days before I/R. Group SC received simvastatin 10 mg/kg via an oro-gastric tube for 3 days. Arterial blood samples were taken at 3 h of reperfusion for blood gas analysis and PaO2 and PaCO2 were recorded. The animals were then sacrificed and the lungs removed immediately for pathologic examination and determination of the wet/dry lung weight ratio (W/D ratio), superoxide dismutase (SOD) activity and polymorphonuclear neutrophil (PMN) count . Hie expression of HO-1 mRNA and protein in lung tissues was detected using RT-PCR and Western blot analysis respectively.Results Alveolar edema, localized pulmonary atelectasis and large amount of PMN infiltration were found in I/R group and were ameliorated in S1, S2, S3 groups. Compared with group S, PaO2 and PaCO2 were significantly decreased in I/R group, W/D ratio and PMN count were increased and SOD activity was significantly decreased in I/R, S1 , S2 groups, and expression of HO-1 mRNA and protein was up-regulated in the other five groups ( P ＜ 0.05). PaO2, PaCO2 and SOD activity were significantly increased, W/D ratio and PMN count were significantly decreased, and HO-1 mRNA and protein expression was up-regulated in S1, S2 and S3 groups as compared with I/R group ( P ＜ 0.05 or 0.01). W/D ratio and PMN count were gradually decreased, SOD activity was gradually increased, and HO-1 mRNA and protein expression was gradually up-regulated in S1, S2 and S3 groups. Conclusion Simvastatin preconditioning has protective effect against lung injury induced by I/R of hind limbs in rats through up-regulation of HO-1 expression in the lung tissues and in a dose-dependent manner.     

PEP-1介导血红素加氧酶-1对大鼠肝脏缺血／再灌注后SOD和MDA及Caspase-3表达的影响

目的：探讨大鼠肝缺血/再灌注（I/R）后PEP-1介导血红素加氧酶-1（HO-1）对肝脏超氧化物歧化酶（SOD）、丙二醛（MDA）及caspase-3的影响。方法制作肝I/R损伤动物模型,SD大鼠随机分为4组,即假手术组（S组）,肝缺血再灌注组（I/R组）、HO-1组、PEP-1-HO-1组。I/R后12 h光镜及电镜下观察肝细胞病理学改变,检测血清ALT的水平、肝组织MDA的含量及SOD的活性,免疫组化染色检测肝组织caspase-3的表达。结果 PEP-1-HO-1组血清ALT、肝组织MDA变化幅度明显低于I/R组,肝组织SOD明显高于I/R组（P ＜0．05）。在电镜下观察,l/R组肝小叶结构紊乱,肝窦淤血,肝细胞水肿变性,肝细胞片状坏死。HO-1组和PEP-1-HO-1组上述改变明显减轻。在I/R组中,caspase-3较S组表达增强,而在HO-1组、PEP-1-HO1组中其表达较I/R组减弱。结论 PEP-1介导HO-1对肝I/R损伤有保护作用,其作用机制可能与减少氧自由基产生、减轻脂质过氧化反应及抑制caspase-3的表达有关。     

细胞穿透肽PEP-1介导的血红素加氧酶-1对大鼠肠缺血再灌注诱发肝损伤的影响

目的 评价细胞穿透肽PEP-1介导的血红素加氧酶-1(HO-1)对大鼠肠缺血再灌注诱发肝损伤的影响.方法 雄性SD大鼠24只,7～9周龄,体重210～260 g,采用随机数字表法,将其分为3组(n=8):假手术组(S组)、肠缺血再灌注组(I/R组)和融合蛋白PEP-1/HO-1组(HO组).采用夹闭肠系膜上动脉45 min,恢复灌注120 min的方法制备大鼠肠缺血再灌注模型.HO组于缺血前30 min经左侧髂静脉注射融合蛋白PEP-1/HO-1 0.5 mg,S组仅分离闭肠系膜上动脉,但不夹闭.于再灌注120 min时,右侧颈总动脉取血样,测定血清AST和ALT的活性;然后处死大鼠,取肝组织,光镜下观察病理学结果,测定MDA含量和SOD活性.结果 与S组比较,I/R组和HO组血清AST和ALT的活性升高,肝组织MDA含量升高,SOD活性降低(P＜0.05);与I/R组比较,HO组血清AST和ALT的活性降低,肝组织MDA含量降低,SOD活性升高(P＜0.05),肝损伤减轻.结论 细胞穿透肽PEP-1介导的HO-1可减轻大鼠肠缺血再灌注诱发的肝损伤.     

血红素氧合酶-1在临床肝移植中肝脏保护作用的研究

目的 研究移植肝脏血红素氧合酶(HO-1)表达水平与缺血再灌注损伤和移植术后肝脏功能的关系.方法 研究28例人类临床原位肝脏移植,根据供肝血红素氧合酶(HO-1)表达的平均值将供肝分为两组:移植前供肝HO-1高表达组和移植前供肝HO-1低表达组.比较两组移植术后血浆AST、ALT水平、胆汁中胆盐含量以及术前术后HO-1 mRNA和蛋白表达情况.结果 再灌注后移植术前HO-1低表达组的HO-1 mRNA表达显著增加,而高表达组HO-1 mRNA表达却有所下降.肝脏移植后,术前HO-1低表达组与高表达组相比,血浆转氨酶显著降低,胆汁中胆盐含量明显高于后者.结论 移植术前HO-1低表达组供肝在再灌注过程中能够进一步诱导HO-1表达,与高表达组供肝相比其所遭受的缺血再灌注损伤较轻,移植术后肝脏功能较好.移植过程中HO-1表达的增强要比移植前HO-1高表达更具有细胞保护作用.免疫荧光染色证实枯否细胞是人类肝脏表达HO-1的主要部位.     

血红素加氧酶-1与自噬在肝脏缺血-再灌注损伤中的研究进展

血红素加氧酶(HO)-1及其酶解产物可通过减轻活性氧自由基损伤、减少细胞凋亡、抑制炎症反应和维持微循环稳定等作用减轻肝脏缺血-再灌注损伤(HIRI)。自噬是细胞利用溶酶体对自身受损细胞器和大分子物质进行生物学降解的过程,被认为是在应激情况下减轻细胞损伤的适应性反应,可有效减少细胞死亡。近年来越来越多的研究表明自噬与HIRI也有着密切的关系。本文回顾了近年来国内外的研究报道,从HO-1与HIRI、自噬与HIRI、HO-1与自噬在HIRI中的相互作用及其机制等方面进行综述。     

血红素加氧酶-1及其与重症胰腺炎之间关系的研究进展

近年来,血红素加氧酶-1(heme oxygenase-1,HO-1)已经成为危重病及其他许多领域的研究热点.HO-1通过分解血红素而产生的一氧化碳,胆绿素和胆红素,及亚铁离子可以减轻炎性介质反应,拮抗细胞氧化性损伤和细胞应激从而发挥细胞保护性作用.而在对重症胰腺炎(severe acute pancreatitis,SAP)的研究中,人们已经认识到拮抗炎症因子的过度释放是提高胰腺炎疗效的重要环节.本文就HO-1及其与SAP之间关系的研究进展做一综述.     

热休克蛋白70和血红素加氧酶-1表达在肾缺血后处理减轻肾缺血再灌注损伤中的作用

目的 评价热休克蛋白70(HSP70)和血红素加氧酶-1(HO-1)表达在肾缺血后处理减轻肾缺血再灌注损伤中的作用.方法健康雄性SD大鼠140只,体重250～280 g,采用随机数字表法,将大鼠随机分为4组(n=35):假手术组(S组)仅开腹,游离双侧肾脏,分离双侧肾蒂不夹团;肾缺血再灌注组(I/R组)夹闭双侧肾蒂缺血45 min,恢复灌注;缺血后处理组(IPo组)夹闭双侧肾蒂45 min,再灌注10 s,缺血10 s,反复3次,恢复灌注;HSP抑制剂槲皮黄酮+缺血后处理组(Q+IPo组)缺血前1 h 腹腔注射槲皮黄酮100 mg/kg,余操作同IPo组.于再灌注即刻(T0)、1、3、6、12、24、48 h(T1～6)时各组随机取5只大鼠抽心脏血后取肾,检测肾组织HSP70、HO-1的mRNA和蛋白表达,T3时抽心脏血,测定血清肌酐(Cr)和尿素氮(BUN)浓度、caspase-3 mRNA的表达,TUNNEL法检测肾组织凋亡细胞,计算凋亡指数(AI),光镜下观察肾组织病理学结果.结果 与S组比较,其余组T3时血清Cr和BUN浓度和AJ升高,caspase-3 mRNA表达上调,各时点HSF70、BO-1的mRNA和蛋白表达上调(P＜0.05);与I/R组比较,IPo组T3时血清Cr和BUN浓度和AI降低,caspase-3 mRNA表达下调,T1～5时HSP70、HO-1的mRNA和蛋白表达上调(P＜0.05);与IPo组比较,Q+IPo组T3时血清Cr和BUN浓度和AJ升高,caspase-3mRNA表达上调,T1～5时HSP70、HO-1的mRNA和蛋白表达下调(P＜0.05).IPo组肾组织病理学损伤较I/R组减轻,Q+IPo组肾组织病理学损伤程度与I/R组相似.结论 HSP70和H0-1表达参与了肾缺血后处理减轻肾缺血再灌注损伤的过程.

Abstract：

Objective To evaluate the role of the expression of heat shock protein 70 (HSP70) and heme oxygenase-1 (HO-1) in the reduction of renal ischemia-reperfusion (I/R) injury by ischemic postconditioning in tats.Methods One hundred and forty healthy male SD rats weighing 250-280 g were randomized into 4 groups ( n = 35 each) : sham operation group (S group) ; I/R group; ischemic postconditioning group (IPo group); quercetin (an inhibitor of HSP) + ischemic postconditioning group (Q + IPo group). Renal I/R was produced by clamping bilateral renal pedicels for 45 min followed by reperfusion. In group S, bilateral kidneys were only exposed through a midline incision but their- pedicels were not clamped. In IPo and Q + IPo groups, 45 min ischemia was followed by three 10 s episodes of ischemia at 10 s intervals for reperfusion and in addition intraperitoneal quercetin 100 mg/kg was injected at 1 h before ischemia in group Q + IPo. Blood samples from hearts were obtained at 0, 1, 3, 6, 12, 24 and 48 h of reperfusion (T0-6) and the rats were then sacrificed and kidneys removed to detect the expression of HSP70 and HO-1 mRNA and protein in renal tissues. The blood samples obtained at T3 were used to determine serum creatinine (Cr) and urea nitrogen (BUN) concentrations and the expression of caspase-3 mRNA . The apoptosis in the renal tissues was detected using TUNEL and apoptotic index ( AI) was calculated. Microscopic examination was performed with light microscope. Results Compared with group S, the serum Cr and BUN concentrations and AI were significantly increased at T3,the expression of caspase-3 mRNA was up-regulated at T3, and the expression of HSP70 and HO-1 mRNA and protein was up-regulated at T0-6 in the other groups (P ＜ 0.05) . Compared with group I/R, the serum Cr and BUN concentrations and AI were significantly decreased at T3, the expression of caspase-3 mRNA was down-regulated at T3, and the expression of HSP70 and HO-1 mRNA and protein was up-regulated at T1-5 in group IPo ( P ＜ 0.05) . Compared with group IPo, the serum Cr and BUN concentrations and AI were significantly increased at T3, the expression of caspase-3 mRNA was up-regulated at T3, and the expression of HSP70 and HO-1 mRNA and protein was down-regulated at T1-5, in group Q + IPo ( P ＜ 0.05) . The microscopic examination showed that the renal I/R injury was significantly attenuated by ischemic postconditioning and the degree of injury in group IPo was similar to that in group I/R. Conclusion The expression of HSP70 and HO-1 is involved in the reduction of renal I/R injury by ischemic postconditioning in rats.     

上调肝脏HO-1抑制肥大细胞脱颗粒减轻大鼠肝脏缺血再灌注损伤

目的 探索HO-1对肝脏缺血再灌注损伤中肥大细胞脱颗粒的影响。方法 将20只SD大鼠随机分成4组：假手术组（Sham组）,缺血再灌注损伤组（I/RI组）,HO-1诱导剂钴原卟啉组(CoPP组,术前24h给予CoPP,5 mg/kg)及HO-1抑制剂锌原卟啉组(ZnPP组,术前24h给予ZnPP,20 mg/kg)。建立大鼠缺血再灌注损伤模型,各组于再灌注后2h收集标本。RT-PCR检测肝脏组织HO-1 mRNA表达,Western blot检测肝脏组织HO-1蛋白表达;测定血清中ALT、AST水平;肝脏组织甲苯胺蓝染色检测肥大细胞脱颗粒数量,HE染色评价肝脏组织损伤情况。结果 与Sham组相比,I/RI组、CoPP组、ZnPP组大鼠组织HO-1 RNA和蛋白表达增加,血清ALT、AST水平升高,肥大细胞脱颗粒数量增多,肝脏细胞损伤加重。CoPP组与I/RI组相比,HO-1 mRNA和蛋白表达增加,血清ALT、AST水平减低,肥大细胞脱颗粒数量减少,肝细胞损伤减轻。ZnPP组与I/RI组相比,HO-1 mRNA和蛋白表达减少,血清ALT、AST水平升高,肥大细胞脱颗粒数量增多、肝细胞损伤严重。组间比较差异具有统计学意义(P＜0.05)。结论 HO-1过表达能减轻肝脏I/RI,其机制可能与抑制肝脏组织中肥大细胞脱颗粒有关。     

大鼠原位肝移植中血红素加氧酶-1表达对BCL-2和VCAM-1影响

目的 探讨血红素加氧酶-1(HO-1)对大鼠原位肝移植后BCL-2和VCAM-1的影响.方法 建立大鼠原位肝移植排斥模型,将大鼠分为三组:对照组(A组),ZnPP组(B组)和CoPP组(C组).三组分别于3 d和7 d处死,取肝脏标本.荧光实时RT-PCR检测HO-1及VCAM-1的表达,免疫组化方法(SP)检测BCL-2的表达.结果 (1)HO-1在C组表达明显升高,B组不明显,A组介于两组之间.(2)VCAM-1在3 d时C组与A或B组相比有明显统计学意义(P＜0.05),A组与B组相比无统计学意义(P＞0.05).7 d时B组与C组相比有明显统计学意义(P＜0.05),A组与B或C组相比无统计学意义(P＞0.05).(3)BCL-2免疫组化染色阳性面积百分率(-x±S,%)统计学分析显示,3 d和7 d时C组与A或B组相比较有明显的统计学意义(P＜0.05),A组和B组相比无统计学意义(P＞0.05).结论 HO-1过表达上调BCL-2的表达,抑制VCAM-1的表达.     

血红素加氧酶-1在脓毒症大鼠炎性反应中的作用

目的 评价血红素加氧酶-1(HO-1)在脓毒症大鼠炎性反应中的作用.方法 雄性Wistar 大鼠72只,10～ 14周龄,体重250～ 300 g,以盲肠结扎穿孔法(CLP)建立脓毒症模型.采用随机数字表法,将大鼠随机分为4组(n=18):对照组(C组)、脓毒症组(CLP组)、钴原卟啉Ⅸ组(Co组)及锌原卟啉Ⅸ组(Zn组).于术后6、12、24 h(T1-3)时随机取6只大鼠采集血样,采用ELISA法检测血清TNF-α、IL-6和高迁移率族蛋白B1(HMGB1)的浓度.于T3时采集血样后取肺组织,采用RT-PCR法测定肺组织HMGB1 mRNA表达.另取40只相同条件的Wistar大鼠,按照上述方法分组,绘制生存曲线.结果 与C组比较,CLP组、Co组和Zn组血清TNF-α、IL-6和HMGB1浓度升高,肺组织HMGB1 mRNA表达上调,生存率降低(P＜0.05).与CLP组比较,Co组血清TNF-α、IL-6和HMGB1浓度降低,肺组织HMGB1 mRNA表达下调,生存率升高(P＜0.05),Zn组上述指标差异无统计学意义(P＞0.05).结论 诱导HO-1表达有助于减轻脓毒症大鼠炎性反应.     

大鼠血红素加氧酶-1基因转染对大鼠离体心肌缺血再灌注心肌细胞凋亡的影响

目的 观察重组腺相关病毒介导大鼠血红素加氧酶-1基因转染对大鼠离体心肌缺血再灌注心肌细胞凋亡的影响.方法 雄性SD大鼠30只随机分成3组,对照组(C组,n=6),缺血再灌注组(I/R组,n=12),腺相关病毒介导血红索加氧酶-1基因组(A-r组,n=12).基因转染3个月后,建立大鼠离体心脏Langendorff灌流模型,C组持续灌注100 min,其他各组均平衡15 rain,停灌40min与再灌注45 min,记录冠脉流量(CF),测定冠脉流出液肌酸激酶(CK)活性,测定再灌注后45 min时的心肌心肌组织超氧化物岐化酶(SOD)活性活性及丙二醛(MDA)含量,同时取每组大鼠心肌检测梗死面积、心肌细胞凋亡率以及心肌组织bax、bcl-2蛋白表达量.结果 离体心肌Langendorff灌注后,与I/R组比较,A-r组在复灌后冠脉流出液CK活性降低(P＜0.01),复灌后45min后心肌MDA含量降低(P＜0.01),SOD活性增高(P＜0.01),梗死面积较小(P＜0.01),心肌组织bax表达量、心肌细胞凋亡率均显著下降,心肌组织bel-2表达量明显增加(P＜0.01).结论 重组腺相关病毒血红素加氧酶1基因转染心肌后,可抑制离体缺血再灌注心肌细胞凋亡和增强心肌抗氧化能力,对大鼠离体心肌缺血再灌注损伤有显著保护作用.     

血红素氧合酶-1在HO-1/Luc小鼠原位肝脏移植模型中的表达

目的 应用活体生物萤光成像技术(BLT)无创定量检测血红素氧合酶-1(HO-1)在活体动物肝脏移植模型中的时空表达.方法 建立小鼠原位肝脏移植模型(HO-1/Luc转基因小鼠8只,wildtype小鼠40只),使用活体生物萤光成像技术连续检测HO-1在移植术后HO-1/Luc转基因小鼠中的转录.应用逆转录-聚合酶链反应(RT-PCR)检测HO-1 mRNA水平,免疫组织化学方法 (IHC)行HO-1的表达定位.结果 移植肝脏发出的萤光信号最早于移植后1 h被检测到(P<0.05),随后信号不断增强在6 h达到峰值.与移植术前比较,除0、48 h外信号差异均有统计学意义(P<0.05).移植后48 h信号衰减至基础水平.与移植术前比较,RT-PCR方法 测得HO-1 mRNA于0～9 h均显著升高(P<0.05),其中于3 h达到峰值,12 h降至基础水平.IHC证实肝脏细胞是移植后HO-1表达上调的主要部位.结论 活体生物萤光成像技术可实时定量检测肝脏移植后HO-1的表达.     

血红素加氧酶-1(HO-1)基因转染治疗急性胰腺炎的研究展望

血红素加氧酶-1(heme oxygenase-1,HO-1)通过分解血红素产生一氧化碳、胆绿素、胆红素及亚铁离子,可以减轻炎性介质反应,拮抗细胞氧化损伤和细胞应激从而发挥细胞保护作用.基因转染的方法能够维持目的基因的长时间表达,可以调节细胞因子的免疫反应.HO-1基因转染已在其他疾病的治疗研究中取得良好成果,本文针对...     

活化蛋白-1在大鼠内毒素性肺损伤时血红素加氧酶-1上调中的作用

目的 评价活化蛋白-1(AP-1)在大鼠内毒素性肺损伤时血红素加氧酶-1(HO-1)上调中的作用.方法 健康清洁级雄性SD大鼠48只,体重200 ～ 220 g,2.5 ～ 3.0月龄,采用随机数字表法,将其随机分为4组(n=12):正常对照组(C组)腹腔注射0.1％二甲基亚砜(姜黄素溶媒)0.5 ml,30 min时股静脉注射生理盐水(LPS溶媒)0.5 ml;内毒素性肺损伤组(ALI组)腹腔注射0.1％二甲基亚砜0.5ml,30 min时股静脉注射10 mg/kg LPS0.5 ml;姜黄素+内毒素性肺损伤组(Cur+ ALI组)腹腔注射20mg/kg姜黄素0.5 ml,30 min时股静脉注射10 mg/kg LPS 0.5 ml;姜黄素组(Cur组)腹腔注射姜黄素20mg/kg,30 min时股静脉注射生理盐水0.5 ml.静脉注射LPS 6 h时处死大鼠取肺组织,行病理学评分,测定MDA含量和SOD活性;采用Western blot法测定HO-1和AP-1表达;采用荧光定量PCR法测定HO-1 mRNA表达.结果 与C组比较,ALl组和Cur +ALI组肺组织病理学评分和MDA含量升高,SOD活性降低,HO-1 mRNA、HO-1和AP-1表达上调(P＜0.05),Cur组上述各指标差异无统计学意义(P＞0.05);与ALl组比较,Cur+ ALl组肺组织病理学评分和MDA含量升高,SOD活性降低,HO-1mRNA、HO-1和AP-1表达下调(P＜0.05).结论 内毒素性肺损伤时HO-1上调的机制与转录因子AP-1活化有一定关系.