高迁移率族蛋白B1在大鼠移植肺缺血再灌注损伤中的作用

目的:探讨高迁移率族蛋白B1(HMGB1)在大鼠移植肺缺血再灌注损伤中的作用。方法:BALB/c小鼠随机分为对照组、10μgHMGB1组和100μg HMGB1组,每组各5只,后两组每只分别腹腔注射重组HMGB1 10μg和100μg。三套管法建立SD大鼠肺移植模型并随机分成单纯移植组、移植加0.1mg抗HMGB1抗体组和移植加1.0mg抗HMGB1抗体组,每组各5只,后两组每只分别腹腔注射抗HMGB1抗体0.1和1.0mg。12h后观察各组肺组织的湿干重比、炎症介质(TNF-α、ICAM-1、IL-6)基因表达、病理表现及动脉血气变化。结果:注射重组HMGB1后尤其100μg HMGB1组小鼠PaO2降低,PaCO2、湿干重比升高,TNF-α、ICAM-1、IL-6基因表达增强,肺组织出现病理改变。抗HMGB1抗体处理后,除ICAM-1基因表达外,肺移植大鼠的其他指标均有明显改善,并与抗HMGB1抗体剂量有关。结论:HMGB1在肺移植缺血再灌注损伤中具有重要作用,是肺移植缺血再灌注损伤防治的潜在靶标。     

针灸预处理对大鼠心肌缺血/再灌注后高迁移率族蛋白B1的影响

目的：探讨针灸预处理对大鼠心肌缺血/再灌注（I/R）后的保护作用以及对高迁移率族蛋白B1（HMGB1）表达的影响。方法选择60只Wistar大鼠,按随机数字表法分为假手术组、心肌I/R模型组、针灸预处理组,每组20只。采用结扎冠状动脉左室支左心耳下缘约0.5 cm处阻断血流10 min后再灌注1 h制备I/R损伤模型;假手术组仅穿线不结扎;针灸预处理组于I/R前7 d给予每日1次电针内关穴20 min,连续治疗7 d。采用苏木素-伊红（HE）染色,光镜下观察心肌组织病理学变化,半定量积分法计算3组心肌组织病理学评分;采用酶联免疫吸附试验（ELISA）检测血浆HMGB1、肿瘤坏死因子-α（TNF-α）、心肌肌钙蛋白T（cTnT）的含量,逆转录-聚合酶链反应（RT-PCR）、蛋白质免疫印迹试验（Western Blot）检测心肌组织HMGB1、单核细胞趋化蛋白-1（MCP-1）、TNF-α的mRNA及蛋白表达水平。结果光镜下可见心肌I/R模型组心肌纤维部分断裂,心肌细胞大片状坏死,边界不清,细胞出现浓缩、破裂、溶解、甚至消失,间质水肿并伴大量炎性细胞浸润;针灸预处理组上述表现较心肌I/R模型组明显减轻。与假手术组比较,心肌I/R模型组HMGB1、TNF-α、cTnT含量和组织病理学评分均明显升高〔HMGB1（μg/L）：9.64±1.16比2.15±.031,TNF-α（μg/L）：91±22比19±5, cTnT（μg/L）：1.50±0.35比0.07±0.03,组织病理学评分（分）：2.5±0.3比0.0±0.0,均P＜0.01〕,HMGB1、MCP-1、TNF-αmRNA和蛋白表达均明显升高（HMGB1 mRNA：1.42±0.16比0.02±0.00,MCP-1 mRNA：0.46±0.06比0.01±0.00,TNF-αmRNA：0.75±0.04比0.03±0.00;HMGB1蛋白：1.08±0.01比0.02±0.01, MCP-1蛋白：0.92±0.03比0.40±0.01,TNF-α蛋白：1.10±0.02比0.35±0.01,P＜0.05或P＜0.01）;与心肌I/R模型组比较,针灸预处理组HMGB1（6.58±0.73）、TNF-α（63±19）、cTnT（1.15±0.31）含量均明显降低（均P＜0.01）,HMGB1、MCP-1、TNF-αmRNA和蛋白表达明显降低（mRNA表达分别为0.74±0.12、0.18±0.02、0.10±0.03,蛋白表达分别为0.40±0.01、0.36±0.02、0.50±0.02,均P＜0.05）,组织病理学评分（1.2±1.0）明显降低（P＜0.01）。结论针灸预处理可减轻大鼠心肌I/R损伤,其机制可能与减轻HMGB1介导的晚期炎症反应有关。     

高迁移率族蛋白B1在肾脏冷缺血再灌注损伤中的作用

背景：缺血再灌注损伤是临床器官移植不可避免的病理生理过程,冷缺血再灌注损伤具有研究肾移植更强的针对性。目的：观察高迁移率族蛋白B1在大鼠肾脏冷缺血再灌注损伤中的作用。方法：将SD大鼠随机分为假手术组、冷缺血再灌注组、丙酮酸乙酯（可显著抑制高迁移率族蛋白B1的合成与释放）治疗组。冷缺血再灌注组和丙酮酸乙酯治疗组制冷缺血再灌注模型前分别经阴茎背静脉注射林格液与丙酮酸乙酯,假手术组将腹腔打开后经阴茎背静脉注射林格液,45min后关闭腹腔。结果与结论：冷缺血再灌注组和丙酮酸乙酯治疗组各时间点肌酐、高迁移率族蛋白B1、肿瘤坏死因子α、核因子κB水平均显著高于假手术组（P〈0.01）,其中冷缺血再灌注组上述指标高于丙酮酸乙酯治疗组（P〈0.01）。表明高迁移率族蛋白参与了肾移植冷缺血再灌注的病理过程,丙酮酸乙酯能够减轻肾脏冷缺血再灌注损伤。     

高迁移率族蛋白B1在肾脏冷缺血再灌注损伤中的作用

背景:缺血再灌注损伤是临床器官移植不可避免的病理生理过程,冷缺血再灌注损伤具有研究肾移植更强的针对性.目的:观察高迁移率族蛋白B1在大鼠肾脏冷缺血再灌注损伤中的作用.方法:将SD大鼠随机分为假手术组、冷缺血再灌注组、丙酮酸乙酯(可显著抑制高迁移率族蛋白B1的合成与释放)治疗组.冷缺血再灌注组和丙酮酸乙酯治疗组制冷缺血再灌注模型前分别经阴茎背静脉注射林格液与丙酮酸乙酯,假手术组将腹腔打开后经阴茎背静脉注射林格液,45 min后关闭腹腔.结果与结论:冷缺血再灌注组和丙酮酸乙酯治疗组各时间点肌酐、高迁移率族蛋白B1、肿瘤坏死因子α、核因子κB水平均显著高于假手术组(P < 0.01),其中冷缺血再灌注组上述指标高于丙酮酸乙酯治疗组(P < 0.01).表明高迁移率族蛋白参与了肾移植冷缺血再灌注的病理过程,丙酮酸乙酯能够减轻肾脏冷缺血再灌注损伤.     

大黄对脓毒症大鼠血浆高迁移率族蛋白B1表达的影响

目的 观察大黄对脓毒症大鼠血清中高迁移率族蛋白B1(HMGB1)、肿瘤坏死因子-α(TNF-α)的影响,揭示大黄治疗脓毒症的机制.方法 采用盲肠结扎穿孔术(CLP)制备脓毒症模型,104只雄性SD大鼠随机分为正常对照组(8 只)、CLP组(48只)、大黄治疗组(48只).分别在术后0、3、8、24、48及72 h活杀大鼠留取血标本,采用酶联免疫吸附法测定TNF-α和HMGB-1的含量.结果 大黄治疗组血浆TNF-α水平在8 h,HMGB1水平在24、48及72 h明显低于CLP组, 差异有统计学意义.结论 大黄通过降低血清中TNF-α和HMGB-1的水平,对脓毒症大鼠具有治疗作用.     

高迁移率族蛋白B1靶向治疗的研究进展

高迁移率族蛋白B1 (HMGB1)为重要炎性因子,其为无菌性炎症与感染相关炎症反应的重要调节者,在感染、炎症及免疫反应中发挥重要作用.HMGB1表达水平可反映机体炎症及组织损伤的严重程度,且与相关疾病的预后密切相关.以HMGB1为治疗靶点,有望为感染、自身免疫性疾病、缺血再灌注损伤(IRI)、移植物抗宿主病(GVHD)等疾病的治疗提供潜在新策略.     

重组高迁移率族蛋白B1致小鼠肝损伤实验研究

目的观察高迁移率族蛋白B1(HMGB1)对小鼠肝脏的病理作用。方法 BALB/c小鼠分为正常对照组、重组HMGB1组和重组HMGB1加抗HMGB1抗体组,每组5只,腹腔注射重组HMGB1和抗HMGB1抗体的剂量分别为100μg/只和500μg/只,注射后24 h,观察血清丙氨酸氨基转移酶、天门冬氨酸氨基转移酶活力、肿瘤坏死因子-α水平以及肝组织形态病理学的变化。另外,采用腹腔注射高剂量的重组HMGB1(500μg),观察小鼠存活情况。结果注射重组HMGB1后,血清丙氨酸氨基转移酶、天门冬氨酸氨基转移酶活力和肿瘤坏死因子-α水平均明显升高(P0.01),肝组织显示凝固性局部坏死、血管充血并伴大量炎性细胞浸润;同时使用抗HMGB1抗体处理后,血清上述指标明显下降(P0.05),组织坏死和炎性细胞浸润明显减轻。5只小鼠注射高剂量重组HMGB1后24、48 h各死亡1只。结论 HMGB1对小鼠肝脏组织细胞具有损伤作用。     

大承气汤对脓毒症大鼠高迁移率族蛋白B1表达的影响

目的 通过观察大承气汤对脓毒症大鼠HMGB1表达的影响,进一步揭示大承气汤治疗脓毒症的分子机制.方法 将96只SD大鼠随机分为假手术组、脓毒症组、低剂量大承气汤组(LDT组)、高剂量大承气汤组(DT组),盲肠结扎穿刺法复制大鼠脓毒症模型.LDT组和DT组分别以不同浓度的大承气汤灌胃,假手术组和脓毒症组以生理盐水对照灌胃.各组大鼠分别于术后2、8、24、48 h四个时间点随机处死6只大鼠,经腹主动脉取血,离心去上清,用ELISA方法检测血浆TNF-α、IL-6、HMGB1的表达.术后24 h,取大鼠末端回肠,用RT-PCR法检测回肠组织HMGB1 mRNA表达,用免疫组织化学方法观察肠组织HMGB1蛋白表达,在光镜下观察大鼠肠组织的病理变化.结果 与假手术组比较,脓毒症组血浆HMGB1在术后8、24、48 h显著增高,脓毒症组回肠组织HMGB1 mRNA及蛋白表达在术后24 h显著增高(P<0.05).与脓毒症组比较,DT组、LDT组血浆HMGB1在术后8、24、48 h显著降低,DT组、LDT组回肠组织HMGB1 mRNA及蛋白表达在术后24 h显著降低(P<0.05).上述指标在LDT与DT组之间比较差异无统计学意义(P>0.05).结论 大承气汤治疗脓毒症的机制可能包括:①抑制肠组织HMGB1的表达,进而保护肠黏膜屏障;②降低血浆HMGB1水平,进而抑制HMGB1的某些受体(如TLR4)介导的炎症介质释放.     

原发性肝癌高迁移率族蛋白B1表达改变及其机制

目的研究原发性肝癌（PHC）组织高迁移率族蛋白B1（HMGB1）表达改变和诱导机制,及PHC患者血清HMGB1水平变化和意义。方法对38例慢性乙肝、32例乙肝后肝硬化、23例继发性肝癌、39例PHC患者和48例健康对照者的血清HMGB1水平进行测定和分析。采用RT-PCR方法,检测11例PHC组织及其癌旁组织中的HMGB1基因表达水平。观察缺氧培养对肝癌细胞株SMMC-7721HMGB1基因表达和胞外释放的影响。结果PHC患者血清HMGB1水平（13．5±6．3μg／L）明显高于健康对照者（3．9±1．4μg／L）,并与AFP水平、TNM分期有关。PHC组织中HMGB1mRNA表达增强。SMMC-7721细胞经缺氧培养3h后,培养上清液中HMGB1含量即见升高,6h后,HMGB1mRNA表达明显增强（P〈O．01）。结论PHCHMGB1表达增强,缺氧为诱导因素。     

高迁移率族蛋白B1在大鼠肝脏热缺血／再灌注损伤中的作用

目的 探讨高迁移率族蛋白B1(HMGB1)在大鼠肝脏热缺血/再灌注(I/R)损伤中的作用.方法 90只Wistar大鼠被随机分为正常对照组、林格液组、丙酮酸乙酯(EP)治疗组.采用夹闭左、右肝蒂使95%肝脏缺血90 min再恢复血流并切除未缺血的尾叶肝脏制模.林格液组和EP治疗组制模前分别经阴茎背静脉注射林格液4 ml或EP 3.26 g/L(溶于林格液4 ml中),于术后2、8、24、48 h取下腔静脉血检测血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)水平,处死动物后检测肝组织HMGB1、肿瘤坏死因子-α(TNF-α)、白细胞介素-10(IL-10)含量.结果 林格液组和EP治疗组血清ALT、AST水平均显著高于正常对照组,EP治疗组各时间点ALT及术后2 h、8 h AST水平均显著低于林格液组(P<0.05或P<0.01).林格液组8、24、48 h肝组织HMGB1水平显著高于正常对照组,EP治疗组仅48 h显著升高;EP治疗组在8 h、24 h时HMGB1水平显著低于林格液组;林格液组和EP治疗组在8 h时TNF-α水平均高于正常对照组,而EP治疗组显著低于林格液组(P<0.05或P<0.01).各组IL-10水平无明显差异.结论 HMGB1参与了肝I/R损伤的病理过程.     

Surfactant replacement in reperfusion injury after clinical lung transplantation

Background: Reperfusion injury remains a significant risk factor in the immediate postoperative course after lung transplantation. We report on our initial clinical experience of surfactant replacement in reperfusion injury after clinical lung transplantation. Methods and results: In 31 consecutive patients, lung (8 single lung, 16 bilateral lung) or heart-lung (7) transplantation was performed. In 6 patients, severe reperfusion injury developed and was treated with continuously nebulized surfactant. Compliance of the allograft increased 40 ± 25 % within 3 h following treatment with surfactant. Alveolar arterial oxygen gradient decreased by 23 ± 11 % after 3 h and by 35 ± 20 % after 6 h. Normal graft function was reestablished within 1–3 days after transplantation. All treated recipients were extubated until the 6th postoperative day. The 30-day mortality for the 31 recipients was 3.3 %, the 1-year survival 84 %. Conclusions: Surfactant replacement may become a clinical method for treatment of reperfusion injury after lung transplantation. Final revision received: 14 April 1999 Accepted: 23 April 1999     

肺移植术后血清HMGB1水平的变化及其临床意义

目的探讨血清高迁移率族蛋白B1（HMGB1）水平在肺移植术后的变化和临床意义。方法肺移植21例,其中术后稳定者10例、术后发生急性排斥反应5例、肺部感染6例。采用酶联免疫吸附试验（ELISA）对19名健康者和肺移植患者术前、术后第1、3、7天的血清HMGB1水平进行检测。结果肺移植术后患者血清HMGB1水平明显升高,急性排斥反应组术后第7天血清HMGB1水平明显高于稳定组（P〈0.01）,肺部感染组术后第3天和术后第7天的血清HMGB1水平均明显高于稳定组和急性排斥反应组。健康对照组血清HMGB1水平明显低于肺移植患者术前水平（P〈0.01）。结论肺移植术后血清HMGB1水平升高,血清HMGB1检测对肺移植术后并发症诊断有较好价值。     

异丙酚对大鼠肾缺血-再灌注后肺损伤的影响

目的:探讨异丙酚对肾缺血-再灌注后肺损伤是否具有保护作用,以及在不同时间点给予异丙酚,其保护作用是否有差异。方法:Wistar大鼠48只,随机分为假手术对照组(C组)、肾缺血-再灌注组(R组),以及异丙酚用药组[分别在缺血前1h(P1组)、缺血即刻(P2组)、再灌注即刻(P3组)和再灌注后1h(P4组)给予异丙酚],共6组,异丙酚输注速度为20mg/(kg·h),共3h。制作肾缺血-再灌注模型。实验结束即刻取动物血样及肺组织,测定超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量以及肺组织形态学变化。结果:与C组相比,R组红细胞及肺组织的SOD活性明显降低(均P<0.01),血清及肺组织的MDA含量明显升高(均P<0.01),光镜下,R组肺组织结构明显受损;与R组相比,各用药组除P4组外,均有所改善;各用药组间变化也有差异。结论:肾缺血-再灌注会引起肺损伤;异丙酚对肾缺血-再灌注所造成的肺损伤有良好的保护作用;给药时间点不同,对异丙酚的保护作用有影响。     

缺氧对巨噬细胞HMGB1释放的影响及其机制

目的观察缺氧对巨噬细胞高迁移率族蛋白B1(HMGB1)释放的诱导作用,并初步探讨其可能的机制。方法采用巨噬细胞株RAW264.7,观察缺氧培养不同时间对培养上清液中HMGB1含量、细胞HMGB1mRNA表达和HMGB1胞内分布的影响,及不同浓度N-乙酰半胱氨酸对HMGB1释放的抑制作用。HMGB1含量和HMGB1mRNA表达水平分别采用酶联免疫吸附试验、实时荧光定量PCR法检测。结果缺氧培养6h后HMGB1mRNA表达水平已显示增强(P0.05),缺氧培养12h后培养上清液中HMGB1含量明显升高(P0.01),且显示HMGB1核浆转移;N-乙酰半胱氨酸对缺氧培养HMGB1释放有剂量依赖性抑制作用。结论缺氧能诱导巨噬细胞释放HMGB1,其机制可能与胞内氧自由基产生有关。     

脓毒症大鼠肺高迁移率族蛋白B1的表达及意义

目的 观察创伤弧菌脓毒症大鼠肺组织HMGB1表达和肺损伤的动态变化,并探讨HMGB1在创伤弧菌脓毒症肺损伤中作用.方法 温州医学院生命科学院实验室,清洁级SD大鼠60只,随机分为正常组(A组,n=10)和创伤弧菌脓毒症组(B组,n=50),采用大鼠左下肢皮下注射创伤弧菌悬液(浓度为6×108cfu/mL,剂量为0.1 mL/100 g)制作大鼠创伤弧菌脓毒症模型),B组于染菌后1、6、12、24、48 h后活杀(各时间点n=10),采用逆转录聚合酶链式反应(RT-PCR)和蛋白免疫印迹(Western blot)分别检测大鼠肺组织HMGB1基因与蛋白的表达,检测肺含水分数和光镜观察肺组织病理变化,数据采用单因素方差分析,并用LSD法进行组间两两比较,P＜0.05为差异有统计学意义.结果 B组染菌后12 h(1.161±0.358,P=0.013)、24 h(1.679±0.235,P=0.000)及48 h(1.258±0.274,P=0.004)大鼠肺组织HMGB1 mRNA表达量较A组(0.652±0.177)明显增高(P＜0.05),并于24 h达到高峰;与A组(0.594±0.190)比较,B组HMGB1蛋白表达量于感染后6 h(1.408±0.567,P=0.026)(P＜0.05)逐渐增加,24 h达到高峰(2.415±1.064,P=0.000);与A组(0.699±0.054)比较,B组大鼠肺含水分数于感染后6 h(0.759±0.030,P=0.001)、12 h(0.767±0.023,P=0.000)、24 h(0.771±0.043,P=0.000)和48 h(0.789±0.137,P=0.000)明显增大(P＜0.05),呈逐渐递增趋势;感染后12 h,大鼠肺内血管明显充血,间质水肿并伴炎性浸润,且逐渐加重,到48 h肺泡腔塌陷明显,肺泡间隔分界不清.结论 大鼠创伤弧菌脓毒症可导致肺脏损伤,HMGB1的表达增加可能是创伤弧菌脓毒症大鼠肺组织损伤的机制之一.

Abstract：

Objective To observe the dynamic changes of high mobility group protein B1 ( HMGB1 )expression in the lung of rats with Vibrio vulnificus sepsis so as to unravel the role of HMGB1 in lung injury.Methods Sixty rats of clean grade were randomly divided into normal control group ( A group, n = 10) and Vibrio vulnificus sepsis group (B group, n =50). Sepsis model was made in rats with subcutaneous injection of Vibrio vulnificus with concentration of 6 × 108 cfu/ml in dose of 0. 1 ml/100 g into left lower limb.The rats of group B were sacrificed 1 h, 6 h, 12 h, 24 h and 48 h after infection for taking lung tissues to detect the water content of lung and to observe the histopathological changes in lung under light microscope.The expression of HMGB1 mRNA and the level of HMGB1 protein in the lungs were detected by RT-PCR and Western blot, respectively. Data were analysed with ANOVA and LSD method for comparison between groups, and P ＜0.05 was considered statistically significant. Results Compared with the group A (0.652±0. 177), the expressions of HMGB1 mRNA in lung of rats of group B were significantly higher in 12 hours (1. 161 ±0.358, P=0.013), 24 hours (1.679 ±0.235, P =0.000) and 48 hours (1.258 ±0.274, P=0.004) and reached the peak in 24 h. Compared with group A (0.594 ±0. 190), the level of HMGB1 protein in rats of group B 6 h after infection ( 1. 408 ± 0. 567, P = 0. 026) was significantly increased (P＜0.05), and it reached peak in 24 h (2.415 ± 1.064, P =0.000) after infection. Compared with group A (0.699 ± 0.054), the lung water contents in rats of group B were significantly increased in 6 h (0.759±0.030, P=0.001), in 12 h (0.767 ±0.023, P =0.000), in 24 h (0.771 ±0.043, P=0.000) and in 48 h (0.789 ±0.137, P=0.000) after infection. Compared with group A, the pathological changes in the lung of rats in group B showed clearly marked pulmonary vascular congestion, interstitial edema and inflammatory cell infiltration, and those changes became more and more serious until alveolar sacs entirely collapsed and the boundaries of the alveolar septa could not be clearly identified in 48 h. Conclusions Vibrio vulnificus sepsis leads to the lung injury of infected rats, and the increase in the expression of HMGB1 mRNA in lung might be one of the mechanisms of lung injury in rats with Vibrio vulnificus sepsis.     

Inhaled nitric oxide (NO) for the treatment of early allograft failure after lung transplantation

Objective: Inhalation of high concentrations of nitric oxide (NO) has been shown to improve gas exchange and to reduce pulmonary vascular resistance in individuals with ischemia-reperfusion injury following orthotopic lung transplantation. We assessed the cardiopulmonary effects of low doses of NO in early allograft dysfunction following lung transplantion. Design: Prospective clinical dose- response study. Setting: Anesthesiological intensive care unit of a university hospital. Patients and participants: 8 patients following a single or double lung transplantation who had a mean pulmonary arterial pressure (PAP) in excess of 4.7 kPa (35 mmHg) or an arterial oxygen tension/fractional inspired oxygen ratio (PaO₂/FIO₂) of less than 13.3 kPa (100 mmHg). Interventions: Gaseous NO was inhaled in increasing concentrations (1, 4 and 8 parts per million, each for 15 min) via a Siemens Servo 300 ventilator. Measurements and results: Cardiorespiratory parameters were assessed at baseline, after each concentration of NO, and 15 min after withdrawal of the agent [statistics: median (25th/75th percentiles: Q1/Q3), rANOVA, Dunnett's test, p < 0.05]. Inhaled NO resulted in a significant, reversible, dose-dependent, selective reduction in PAP from 5.5(5.2/6.0) kPa at control to 5.1(4.7/5.6) kPa at 1 ppm, 4.9(4.3/5.3) kPa at 4 ppm, and to 4.7(4.1/5.1) kPa at 8 ppm. PaO₂ increased from 12.7(10.4/17.1) to 19.2(12.4/26.0) kPa at 1 ppm NO, to 23.9(4.67/26.7) kPa at 4 ppm NO and to 24.5(11.9/28.7) kPa at 8 ppm NO. All patients responded to NO inhalation (either with PAP or PaO₂), all were subject to long-term inhalation (1–19 days). All were successfully weaned from NO and were discharged from the intensive care unit. Conclusion: The present study demonstrates that low-dose inhaled NO may be an effective drug for symptomatic treatment of hypoxemia and/or pulmonary hypertension due to allograft dysfunction subsequent to lung transplantation. Received: 23 February 1998 Accepted: 20 August 1998     

七氟醚对兔肺缺血-再灌注损伤中血红素氧合酶-1的影响

目的探讨七氟醚对兔肺缺血-再灌注损伤的血红素氧合酶-1(heme oxygenase-1,HO-1)的影响。方法健康雄性日本大耳白兔24只,随机分成四组(n=6)。假手术组(S组):开胸游离左肺门后,未行缺血-再灌注处理;缺血-再灌注组(IR组):阻断左肺门45min后松开血管夹再灌注120min;七氟醚-缺血再灌注组(Sev-IR组):先吸入1MAC七氟醚30min后行缺血-再灌注;七氟醚组(Sev-S组):持续吸入1MAC七氟醚30min,未予缺血-再灌注处理。缺血45min,再灌注120min时处死兔,观察各组肺组织HO-1活性,肺组织湿干重比(W/D)、肺泡损伤数(IRA)及电子显微镜观察肺组织超微结构的改变。数据采用单因素方差分析。结果再灌注120min后IR组和Sev-IR组肺组织W/D、IRA值均高于S组(P<0.01),而Sev-IR组的这两个指标较IR组均有明显降低(P<0.01);再灌注120min后IR组和Sev-IR组肺组织HO-1活性分别为(489.86±72.18)和(758.67±111.15)较S组(135.58±36.47)均明显升高(P<0.01);与IR组相比,Sev-IR组肺组织HO-1活性明显升高(P<0.01)。肺组织的电镜结果显示七氟醚使IR诱导的超微结构损伤减轻。上述指标于S和Sev-S组间差异无统计学意义。结论七氟醚可增强肺组织HO-1活性,这是其对肺IR损伤发挥的保护作用的机制之一。     

Lipoxin A4 attenuates the lung ischaemia reperfusion injury in rats after lung transplantation

BackgroundLung ischaemia reperfusion injury (LIRI) is the major cause of primary lung dysfunction after lung transplantation. Lipoxin A4 inhibits the oxidative stress and inflammation. This study aimed to evaluate the potential protective effect of lipoxin A4 on LIRI in rats.MethodsSD (Sprague-Dawley) rats were randomised into the sham, LIRI and LA4 groups. Rats in the sham group received anaesthesia, thoracotomy and intravenous injection of saline, while those in the LIRI or LA4 group received left lung transplantation and intravenous injection of saline or lipoxin A4, respectively. After 24 h of reperfusion, the PaO₂/FiO₂ (Partial pressure of O2 to fraction inspiratory O2), wet/dry weight ratios and protein levels in lungs were measured to assess the alveolar capillary permeability. The oxidative stress response and inflammation were examined. The histological and apoptosis analyses of lung tissues were performed via HE staining (Haematoxylin-eosin staining) and TUNEL assay, respectively. The effects of lipoxin A4 on the endothelial viability and tube formation of hypoxaemia and reoxygenation-challenged rat pulmonary microvascular endothelium cells were determined.ResultsLipoxin A4 significantly ameliorated the alveolar capillary permeability, reduced the oxidative stress and inflammation in transplanted lungs. The histological injury and apoptosis of lung tissues were also alleviated by lipoxin A4. In vitro lipoxin A4 treatment promoted the endothelial tube formation and improved the endothelial viability.ConclusionLipoxin A4 protects LIRI after lung transplantation in rats, and its therapeutic effect is associated with the properties of anti-inflammation, anti-oxidation, and endothelium protection.

Key messages:

1. Lung transplantation is a treatment approach for the patients with lung disease.
2. LIRI is the major cause of postoperative primary lung dysfunction.
3. Lipoxins A4 exhibits strong anti-inflammatory properties.