线粒体抗氧化肽对脓毒性脑病小鼠认知功能和线粒体功能的影响

目的观察线粒体抗氧化肽SS-31对脓毒性脑病小鼠认知功能和线粒体功能的影响。方法成年雄性C57BL/6小鼠60只随机分为四组:假手术+生理盐水组(SN组,n=10)、假手术+SS-31组(SS组,n=10)、盲肠结扎穿孔(CLP)+生理盐水组(CN组,n=20)和CLP+SS-31组(CS组,n=20)。术毕每天腹腔注射5mg/kg SS-31(SS、CS组)或等容生理盐水(SN、CN组),连续6d。术后第7、8天,行条件性恐惧实验;行为学测试后处死小鼠,取海马组织检测活性氧自由基(ROS)及三磷酸腺苷(ATP)水平,提取线粒体检测线粒体电子传递链复合物Ⅰ、Ⅱ、Ⅲ、Ⅳ酶活性及线粒体膜电位(MMP)水平。结果与CN组比较,SN、SS和CS组24h场景实验僵直时间明显增加(P0.05),ROS水平明显降低,ATP、MMP及复合物Ⅰ和Ⅲ酶活性明显升高(P0.05)。与CS组比较,SN和SS组ROS水平明显降低,ATP、MMP及复合物Ⅰ和Ⅲ酶活性明显升高(P0.05)。结论线粒体抗氧化肽SS-31可改善脓毒性脑病小鼠认知功能,其机制可能与减轻线粒体损伤有关。     

瑞芬太尼对腹腔感染脓毒症小鼠肝肺组织诱导型一氧化氮合酶的影响

目的 观察瑞芬太尼对腹腔感染脓毒症小鼠肺组织和肝组织诱导型一氧化氮合酶(iNOS)的影响.方法 采用小鼠盲肠结扎穿孔术制备脓毒症模型,将40只雄性昆明小鼠随机分为假手术组(Sham组)、脓毒症组(CLP组)、瑞芬太尼治疗组(R1组)、瑞芬太尼对照组(R2组).Western blot方法检测肺组织和肝组织iNOS蛋白表达,双抗体夹心酶联免疫吸附法测定肝肺组织白细胞介素(IL)-10和IL-6水平,观察PaO_2、肺组织髓过氧化物酶(MPO)活性、血清ALT和AST活性及电镜下组织超微结构改变.结果 与Sham组比较,CLP组PaO_2显著降低,肺组织和肝组织中iNOS蛋白表达、肺组织MPO活性、ALT和AST活性显著增强(P<0.01),肺组织IL-6和IL-10水平显著增加为649.74±90.47和501.06±57.67(P<0.01),肝组织IL-6和IL-10水平显著增加为341.05±28.73和267.50±41.82,R2组以上指标均无明显变化;与CLP组比较,R1组PaO_2明显增加,iNOS蛋白表达,IL-6和IL-10水平、MPO活性、ALT和AST活性显著降低(P<0.01),电镜显示肺组织和肝组织损伤程度较CLP组略减轻.结论 瑞芬太尼对脓毒症感染致急性肺损伤和肝损伤有保护作用,其机制可能通过抑制iNOS蛋白的表达,降低炎性因子水平有关.     

氢气对脓毒症小鼠急性肺损伤的影响

目的 评价氢气对脓毒症小鼠急性肺损伤的影响.方法 雄性C57BL/6小鼠112只,体重20～25 g,5周龄,采用随机数字表法,将其随机分为4组(n=28):假手术组(A组)、假手术+氢气组(B组)、脓毒症组(C组)和脓毒症+氢气组(D组).采用盲肠结扎穿孔法(CLP)制备脓毒症模型.B组和D组于CLP或假手术后1、6h时分别吸入2％氢气.每组取20只小鼠,记录CLP后7d内生存情况.每组取8只小鼠,于CLP后24 h时处死,采集静脉血样,取肺组织,测定血清及肺组织超氧化物歧化酶(SOD)、过氧化氢酶(CAT)及8-异前列腺素F2α(8-iso-PGF2α)水平,测定支气管肺泡灌洗液(BALF)蛋白浓度和肺组织髓过氧化物酶(MPO)活性,进行肺损伤评分,计算肺湿干重比(W/D).结果 与A组比较,C组生存率、血清和肺组织SOD和CAT活性降低,肺损伤评分、W/D、BALF蛋白浓度、肺组织MPO活性、血清和肺组织8-iso-PGF2α水平升高(P＜0.05);与C组比较,D组生存率、血清和肺组织SOD和CAT活性升高,肺损伤评分、W/D、BALF蛋白浓度、肺组织MPO活性、血清和肺组织8-iso-PGF2α水平降低(P＜0.05).结论 氢气可减轻脓毒症小鼠急性肺损伤,其机制与抑制氧化应激反应有关.     

戊乙奎醚预先给药对大鼠急性肺损伤时NF-κB的影响

目的 探讨戊乙奎醚预先给药对大鼠急性肺损伤(ALI)时NF-κB的影响.方法 雄性SD大鼠35只,体重210～280 g,随机分为5组(n=7):对照组(C组)、ALI组和低、中、高剂量戊乙奎醚组(P1-3组).采用经尾静脉注射内毒素5 mg/kg的方法建立大鼠ALI模型.C组和ALI组经腹腔注射生理盐水0.5 ml,P1～3组分别经腹腔注射戊乙奎醚0.03、0.1和3 mg/kg,30 min后ALI组、P1～3组制备AU模型,C组不制备模型.于静脉注射LPS后4 h时,行血气分析,计算氧合指数;称量肺组织湿重(W)、干重(D),计算W/D;采用比色法测定肺组织髓过氧化物酶(MPO)活性;采用RT-PCR法测定肺组织肿瘤坏死因子-α(TNF-α)mRNA、白细胞介素-1β(IL-1β)mRNA的表达水平;采用ELISA法测定肺组织TNF-α和IL-1β的含量;采用非放射性EMSA法测定肺组织NF-κB活性;采用免疫组织化学法测定肺组织NF-κB的表达水平.结果 与C组比较,其余各组氧合指数降低,肺组织W/D和MPO活性、TNF-α,IL-1β含量及其相应mRNA表达水平、NF-κB活性和表达水平均升高(P<0.05或0.01);与ALI组比较,P1～3组氧合指数升高,肺组织W/D和MPO活性降低,TNF-α、IL-1β含量及其相应mRNA表达水平降低,NF-κB活性和表达水平降低(P<0.05);P1～3组上述指标比较差异无统计学意义(P>0.05).结论 戊乙奎醚预先给药减轻大鼠急性肺损伤的机制可能与抑制NF-κB的活化,下调NF-κB的表达,降低肺组织炎性反应有关.     

核因子-κB和诱导性一氧化氮合成酶在大鼠肝脏小移植物缺血预处理中的作用

目的 探讨核因子-κB(NF-κB)和诱导性一氧化氮合成酶(iNOS)在大鼠肝脏小移植物缺血预处理中的作用。方法雄性SD大鼠随机分成5组：假手术组(A组)；小移植物组(B组)；应用二硫代氨基甲酸吡咯烷(PDTC)的小移植物组(C组)；应用缺血预处理(IPC)的小移植物组(D组)；应用PDTC IPC的小移植物组(E组)。电泳迁移率改变法(EMSA)检测小移植物植入后5个时间点肝脏组织中NF-κB活性，逆转录一聚合酶链反应(RT-PCR)检测iNOS在肝组织中的表达，检测肝脏脂质过氧化产物丙二醛(MDA)和超氧化物歧化酶(SOD)的浓度。结果 A组肝组织中NF-KB的活性很弱。B、C、D、E4组NF-κB变化趋势相仿，一般在恢复灌注后即活化，峰值出现在3h(IPC)；活化峰值强度B组＞C组＞D组＞E组。再灌注2h后B、C、D、E组肝组织中iNOS表达水平开始升高，6h达到峰值，其值B组＞D组＞c组＞E组(P＜0．05)，随后逐渐下降；至12h仍高于对照组的水平。MDA的结果变化和iNOS表达水平变化类似，SOD呈相反变化。结论 NF-κB在大鼠肝脏缺血预处理小移植物中起重要作用。抑制供肝组织中NF-κB的活性可显著降低再灌注早期供肝组织中iNOS表达和氧自由基水平．并有效改善供肝的再灌注损伤。针对NF-κB的靶向治疗可能是供肝保护的一条新途径。     

盐酸戊乙奎醚对内毒素血症大鼠全身炎性反应的影响

目的 探讨盐酸戊乙奎醚对内毒素血症大鼠血清TNF-α、诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS),肺组织NF-κB亚基p65亲和肽(NF-κB p65)含量的影响. 方法 健康SD大鼠60只,采用随机数字表法分为5组(每组12只):对照组(C组)、内毒素组(LPS组)、LPS+低剂量盐酸戊乙奎醚组(P1组)、LPS+中剂量盐酸戊乙奎醚组(P2组)、LPS+高剂量盐酸戊乙奎醚组(P3组).LPS组腹腔注射内毒素8 mg/kg制备内毒素血症模型,C组给予等量生理盐水,P1组～P3组给予相应剂量的盐酸戊乙奎醚5 min后注射LPS,给药6h后处死动物取标本.采用ELISA检测血清TNF-α含量,采用比色法测血清iNOS活力,Western blot检测肺组织NF-κB p65含量,并观察肺组织病理学改变. 结果 与C组比较,LPS组、P1组、P2组、P3组血清TNF-α含量显著升高(P＜0.05),iNOS活力水平明显升高(P＜0.05),肺组织NF-κB p65表达明显增高(P＜0.05);与LPS组比较,P2组、P3组血清TNF-α、iNOS活力水平和肺组织NF-κB p65含量均下降(P＜0.05),而P1组TNF-α、iNOS活力和NF-κB p65表达水平,差异无统计学意义(P＞0.05);P2组、P3组间各指标表达水平比较,差异无统计学意义(P＞0.05);P2组、P3组肺组织病理学损伤程度明显轻于LPS组(P＜0.05). 结论 盐酸戊乙奎醚可能通过下调TNF-α、NF-κB p65的表达,减少iNOS活化来抑制内毒素休克大鼠全身炎症反应.     

不同剂量瘦素对大鼠机械通气肺损伤的影响

目的评价不同剂量瘦素对大鼠机械通气肺损伤的影响。方法健康清洁级SD雄性大鼠48只,6～8周龄,采用随机数字表法分为四组:气管切开保留自主呼吸的假手术组(A组)、机械通气模型组(B组)、瘦素10μg/kg组(C组)和瘦素50μg/kg组(D组),每组12只。采用10%水合氯醛3.5 ml/kg麻醉大鼠,疼痛反射消失后C组腹腔注射瘦素10μg/kg,D组腹腔注射瘦素50μg/kg,A、B组腹腔注射等容量生理盐水,注射后即刻进行气管切开,插管机械通气。A组气管插管后保留自主呼吸,B、C、D组机械通气建立VILI模型,参数设置:V_T 20 ml/kg,RR 80次/分,I∶E 1∶1,FiO_2 21%,PEEP 0 mmHg,通气时间4 h。分别于基础状态、通气结束时抽取股动脉血进行血气分析。通气结束后放血处死大鼠,在4℃下取肺组织并收集支气管肺泡灌洗液(bronchoalveolar lavage fluid, BALF),光镜下进行中性粒细胞计数,采用ELISA法测定TNF-α、IL-6、IL-1β浓度;取肺组织称重,计算肺湿干重比(W/D);观察肺组织病理改变并进行病理评分;采用Western blot检测肺组织研磨液中NF-κB p65含量。结果与A组比较,B、C、D组W/D、肺损伤评分、BALF中性粒细胞计数、TNF-α、IL-6、IL-1β浓度及肺组织NF-κB p65含量明显升高(P0.01)。与B组比较,C、D组BALF中性粒细胞计数、TNF-α、IL-6、IL-1β浓度及肺组织肺损伤评分、NF-κB p65含量明显降低(P0.05)。与C组比较,D组BALF中性粒细胞计数、TNF-α、IL-6、IL-1β浓度及肺组织肺损伤评分、NF-κB p65含量明显降低(P0.05)。结论瘦素可降低大鼠机械通气肺损伤中炎性因子的表达水平,减轻肺损伤,50μg/kg较10μg/kg作用明显。     

核因子-κB在体外循环后肺损伤发病机制中的作用

目的探讨核因子-κB(NF-κB)在体外循环后肺损伤发病机制中的作用和甲基强的松龙预处理对肺损伤的防治作用。方法健康雄性家犬12条,随机均分为两组:甲基强的松龙组(M组)和对照组(C组)。持续检测ECG、MAP和平均肺动脉压(MPAP),体外循环前(T0)、主动脉开放30 min(T2)9、0 min(T3)分别测定CVP、肺动脉楔压(PCWP)、心排出量(CO)、肺顺应性和动脉血气并计算肺血管阻力(PVR)、心脏指数(CI)和肺泡-动脉氧分压差(PA-aDO2)。T0、主动脉开放前(T1)、T2、T3分别:(1)采取动脉血测定肿瘤坏死引子α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8);(2)取适量右上肺组织测定肺组织湿干重比(W/D)、髓过氧化物酶活性(MPO)变化、细胞核内NF-κB p65亚单位的活性变化及肺病理学改变。结果(1)两组T1、T2、T3和T0相比,NF-κB活性、TNF-α、IL-1β、IL-6、IL-8、MPO和W/D均显著增加(P<0.01),T1、T2、T3M组和C组相比NF-κB活性、TNF-αI、L-1βI、L-6I、L-8、MPO和W/D均显著降低(P<0.01);(2)病理学改变:T1、T2、T3和T0相比两组均出现肺间质中性粒细胞浸润,肺间隔增宽,肺泡腔有渗出并且C组较M组改变明显;(3)血液动力学改变:MAP、CI在组内及两组之间各时点均无显著变化,T2、T3C组PVR较T0明显升高(P<0.05),而M组无显著变化;(4)肺功能:两组T2、T3较T0PA-aDO2显著增加(P<0.01),M组T2、T3较C组PA-aDO2明显降低(P<0.05);C组T2、T3肺顺应性较T0明显降低(P<0.05),而M组无明显变化,PaCO2无明显变化。结论体外循环可引起肺组织NF-κB的活化和细胞因子的显著增加,发生肺损伤;甲基强的松龙可抑制NF-κB的活性,减轻肺损伤。     

洛沙坦对糖尿病小鼠机械通气相关肺损伤的影响

目的 评价洛沙坦对糖尿病小鼠机械通气相关肺损伤的影响.方法 SPF级雌性C57/BL6小鼠48只,10 ～ 12周龄,体重20 ～ 25 g,采用随机数字表法,将小鼠随机分为3组(n=16)∶对照组(C组)、糖尿病机械通气组(DM组)和洛沙坦组(L组).DM组和L组采用腹腔注射链脲佐菌素150mg/kg的方法制备糖尿病模型.取糖尿病模型制备成功的小鼠进行机械通气4h,吸入氧浓度50％,通气频率70次/min,潮气量15 ml/kg,PEEP 2 cm H2O.L组于机械通气前30 min腹腔注射洛沙坦30mg/kg.机械通气4h时采集颈动脉血样,测定PaO2,随后处死,取肺组织,测定湿/干重比(W/D比)、髓过氧化物酶(MPO)活性和肺微血管通透性;采用实时荧光定量RT-PCR法测定血管紧张素Ⅱ(AngⅡ)受体AT1 mRNA的表达水平;ELISA法测定AngⅡ的含量;Western blot法测定细胞核内NF-κB p65的表达水平.结果 与C组比较,DM组和L组PaO2降低,肺组织W/D比、MPO活性、肺微血管通透性和AngⅡ含量升高,AT1 mRNA和NF-κB p65表达上调(P＜0.05);与DM组比较,L组PaO2升高,肺组织W/D比、MPO活性、肺微血管通透性和Ang Ⅱ含量降低,AT1 mRNA和NF-κB p65表达下调(P＜0.05).结论 洛沙坦通过抑制AT1受体和Ang Ⅱ水平,从而改善肺微血管通透性和抑制NF-κB活化,减轻糖尿病小鼠机械通气相关肺损伤.     

重组人红细胞生成素对脓毒症大鼠肾脏的保护作用

目的 探讨重组人红细胞生成素(rHuEPO)对盲肠结扎穿孔术(CLP)所致大鼠脓毒症急性肾损伤(AKI)的保护作用.方法 260只雄性SD大鼠随机分为正常对照组、假CLP组(假手术组)、CLP组(脓毒症组)和rHuEPO大、中、小剂量组.采用CLP复制脓毒症模型,rHuEPO大、中、小剂量组造模后即刻分别静脉推注rHuEPO 5000 U/kg、1000 U/kg、500 U/kg.假CLP组、CLP组、rHuEPO大、中、小剂量组按注药后2h、6h、12 h、24 h、36 h时点分成5个亚组,分别在各时点按随机原则处死大鼠.采用ELISA法测定血清中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、肾损伤分子1(KIM-1)、诱导型一氧化氮合酶(iNOS)的水平;免疫组化法检测大鼠肾组织NF-κB蛋白表达;光镜和透射电镜观察肾组织形态学变化.结果 与CLP组相比,rHuEPO大剂量组大鼠各时间点血清Scr、BUN、TNF-α、IL-6、KIM-1、iNOS的含量及肾组织NF-κB蛋白表达水平均显著降低,差异有统计学意义(P＜0.05);肾脏病理改变好转.rHuEPO中、小剂量组肾脏病理改变未见好转.结论 rHuEPO能够降低脓毒症大鼠血清TNF-α、IL-6、iNOS、KIM-1水平进而调节炎性反应及肾功能,对脓毒症所致AKI具有一定的保护作用.     

胆碱酯酶抑制剂他克林对兔内毒素性脑损伤的影响

目的 评价胆碱酯酶抑制剂他克林对兔内毒素性脑损伤的影响.方法 健康成年雄性新西兰大白兔21只,体重1.7～2.3 ks,采用随机数字表法,将其随机分为3组(n＝7):假手术组(S组)脑池内注射生理盐水；内毒素组(LPS组)脑池内注射内毒素200 μg/kg;胆碱酯酶抑制剂组(THA组)脑池内注射内毒素200 μg/kg和盐酸他克林150 μg/kg.注射后4h时处死动物,取脑组织,提取细胞核蛋白,采用Western blot法测定NF-κBp65表达,ELISA法测定血浆、脑脊液和脑组织TNF-α水平,采用分光光度法测定脑组织髓过氧化物酶(MPO)活性,称重后计算湿干重比.结果 与S组比较,LPS组和THA组NF-κB p65表达上调,血浆、脑脊液和脑组织TNF-α水平、MPO活性和湿干重比升高(P<0.05)；与LPS组比较,THA组NF-κB p65表达下调,血浆、脑脊液和脑组织TNF-α水平、MPO活性和湿干重比降低(P<0.05).结论 胆碱酯酶抑制剂他克林可通过抑制局部炎性反应减轻兔内毒素性脑损伤.     

盐酸戊乙奎醚预先给药对内毒素性急性肺损伤大鼠肺组织NF-κB mRNA表达及SOD活性的影响

目的 探讨盐酸戊乙奎醚预先给药对内毒素性急性肺损伤大鼠肺组织NF-κB mRNA表达及SOD活性的影响.方法 健康雄性SD大鼠32只,月龄2月,体重230～280 g,随机分为4组(n=8),对照组(C组)腹腔和尾静脉均注射生理盐水1 ml/kg;急性肺损伤组(ALI组):腹腔注射生理盐水1 ml/kg,30 min后经尾静脉注射LPS 5 mg/kg;盐酸戊乙奎醚低剂量组(LP组)、高剂量组(HP组)分别腹腔注射盐酸戊乙奎醚0.3和1 mg/kg,30 min后经尾静脉注射LPS 5 mg/kg.静脉注射生理盐水或LPS后6 h时,取肺组织,检测NF-κB mRNA的表达、TNF-α和MDA的含量和SOD活性,计算肺组织湿/干重比(W/D)及含水量,观察肺组织病理学结果.结果 与C组比较,ALI组、LP组和HP组肺组织NF-κB mRNA表达上调,TNF-α及MDA含量升高,SOD活性降低,W/D和肺组织含水量升高(P＜0.05);与ALI组比较,LP组和HP组肺组织NF-κB mRNA表达下调,TNF-α及MDA含量降低,SOD活性升高,W/D和肺组织含水量降低(P＜0.05);与LP组比较,HP组肺组织NF-κB mRNA表达下调,TNF-α及MDA含量降低,SOD活性升高,W/D和肺组织含水量降低(P＜0.05).LP组和HP组肺组织病理学损伤较ALI组减轻.结论 盐酸戊乙奎醚预先给药减轻大鼠内毒素性急性肺损伤的机制可能与下调肺组织NF-κB mRNA表达,降低肺局部炎性反应,增强机体抗氧化能力有关.     

Effect of TLR-4 and HO-1 on acute lung injury induced by hemorrhagic shock in mice

Objective： To examine whether TLR-4 has an ettect on hemorrhage induced changes in lung, and to investigate the change of heme oxygenase-1 （HO-1） on acute lung injury （ALl） induced by hemorrhagic shock in mice.
Methods： Forty-eight male mice, including C3H/HeN mice and C3H/HeJ mice, were randomly divided into sham group （n=12）, hemorrhagic shock group with twelve mice in each phase. Blood pressure （BP） was monitored continuously by attaching carotid artery catheter to a strain gauge pressure transducer/ polygraph. Arterial blood samples were taken for blood gas analysis. A mouse model of non-lethal hemorrhagic shock and resuscitation was used to observe pulmonary myeloperoxidase （MPO） activity and wet/dry weight ratio （W/D）. The expression of HO- 1 was observed by means of RT-PCR and immunohistochemistry. IL-6 and IL-10 in lung tissue homogenate were assayed by enzyme-linked immunosorbent assay （ELISA）. The pulmo- nary pathologic changes were observed under electron microscope and light microscope.
Results： Compared with sham group, the expression of HO- 1 in lung tissue was significantly higher in Hem 24 h and Hem 48 h of C3H/HeN mice （P〈0.01）. The expression of HO-1 mRNA and the levels of IL-6, IL-10 and MPO in lung tissue were markedly increased in Hem 24 h （P〈0.01 or P〈0.05）; Compared with C3H/HeN mice, the expression of HO- 1 rnRNA and the levels of IL-6 and IL-10 in C3H/HeJ mice significantly decreased in Hem 24 h and Hem 48 h （P〈0.01 or P〈O.05）, and the W/D, MPO in C3H/HeJ mice were obvi- ously lower in Hem 24 h （P〈0.05）. The injuries of lung tissues after hemorrhagic shock have been demonstrated by histological examination with electron microscope and light microscope.
Conclusions： TLR-4 and HO-1 might modulate the bal- ance of pro- and anti-inflammatory processes in inflamma- tory reaction of hemorrhagic shock-induced ALl, and the activation of Toll-like receptor might induce the transcrip- tion activity of HO- 1, which may play a k     

曲马多对致痛小鼠血清核因子-κB及白细胞介素-2的影响

目的 研究曲马多对致痛小鼠血清核因子-κB(NF-κB)和白细胞介素-2(IL-2)的影响.方法 112只BALB/c雄性小鼠.体重25～30 g,随机均分为网组:曲马多组(T组,50 mg/kg曲马多)、吗啡组(M组,5 mg/kg吗啡)和假给药组(S组,生理盐水)、空白对照组(C组).各组在给药后1 h(T_1)、3 h(T_2)、12 h(T_3)、24 h(T_4)时眶静脉采血,用酶联免疫吸附法(ELISA)测定血清NF-κB及IL-2的含量,热板法测痛阈.结果 与S组比较,T组和M组的各时点痛阈增高(P<0.05).NF-κB:与C组比较,T_4时T组升高(P<0.05),T_3、T_4时M组和S组降低(P<0.05);与S组比较,T_2～T_4时T组升高;与M组比较,T_2～T_4时T组升高(P<0.05).T_1～T_3时T组IL-2含量高于M组(P<0.05).结论 吗啡对福尔马林致痛小鼠血清NF-κB和IL-2有抑制作用.曲马多对其具有增强作用,提示曲马多可能对疼痛所致的免疫抑制有改善作用.     

Curcumin protects against sepsis-induced acute lung injury in rats

The present study aimed to investigate the effect of curcumin on sepsis-induced acute lung injury (ALI) in rats, and explore its possible mechanisms. Male Sprague-Dawley rats were randomly divided into the following five experimental groups (n = 20 per group): animals undergoing a sham cecal ligature puncture (CLP) (sham group); animals undergoing CLP (control group); or animals undergoing CLP and treated with vehicle (vehicle group), curcumin at 50 mg/kg (low-dose curcumin [L-Cur] group), or curcumin at 200 mg/kg (high-dose curcumin [H-Cur] group).At 6, 12, 24 h after CLP, blood, bronchoalveolar lavage fluid (BALF) and lung tissue were collected. The lung wet/dry weight (W/D) ratio, protein level, and the number of inflammatory cells in the BALF were determined. Optical microscopy was performed to examine the pathologic changes in lungs. Myeloperoxidase (MPO) activity, malondialdehyde (MDA) content, as well as superoxidase dismutase (SOD) activity were measured in lung tissues. The expression of inflammatory cytokines, tumor necrosis factor-alpha (TNF-α), interluekin-8 (IL-8), and macrophage migration inhibitory factor (MIF) were determined in the BALF. Survival rates were recorded at 72 h in the five groups in another experiment. Treatment with curcumin significantly attenuated the CLP-induced pulmonary edema and inflammation, as it significantly decreased lung W/D ratio, protein concentration, and the accumulation of the inflammatory cells in the BALF, as well as pulmonary MPO activity. This was supported by the histopathologic examination, which revealed marked attenuation of CLP-induced ALI in curcumin treated rats. In addition, curcumin significantly increased SOD activity with significant decrease in MDA content in the lung. Also, curcumin caused down-regulation of the inflammatory cytokines TNF-α, IL-8, and MIF levels in the lung. Importantly, curcumin improved the survival rate of rats by 40%-50% with CLP-induced ALI. Taken together, these results demonstrate the protective effects of curcumin against the CLP-induced ALI. This effect can be attributed to curcumin ability to counteract the inflammatory cells infiltration and, hence, ROS generation and regulate cytokine effects.     

盐酸戊乙奎醚通过NF-κB信号通路对胎鼠脑缺血-再灌注损伤的影响

目的研究盐酸戊乙奎醚对宫内窘迫致胎鼠脑缺血-再灌注(IR)损伤的保护作用。方法足月胎鼠80只,体重4.52~4.81 g,采用随机数字表法分为四组:假手术组(S组)、盐酸戊乙奎醚对照组(S+P组)、IR组和盐酸戊乙奎醚治疗组(IR+P组),每组20只。采用钳夹孕鼠两侧子宫角血管的方法建立宫内窘迫模型,IR+P组孕鼠建模前30 min肌注盐酸戊乙奎醚2 mg/kg,S组在假手术前予孕鼠肌注等量生理盐水,S+P组在假手术前予孕鼠肌注等量盐酸戊乙奎醚。再灌注12 h后,剖宫取出胎鼠并断头处死,行外周血血气分析;采用TTC染色法观察胎鼠脑梗死体积并计算脑梗死体积百分比;采用HE染色观察胎鼠脑组织病理改变;采用ELISA法检测脑组织中TNF-α及IL-6的浓度;采用RT-PCR法检测脑组织中NF-κB mRNA表达;采用Western blot法检测胎鼠脑组织中NF-κB p65蛋白表达。结果 IR组和IR+P组p H和Pa O2明显低于S组和S+P组,IR+P组p H和Pa O2明显高于IR组(P0.05)。IR组和IR+P组Pa CO2、Lac、脑梗死体积及梗死体积百分比、TNF-α及IL-6浓度、NF-κB mRNA表达水平、NF-κB p65蛋白水平均明显高于S组和S+P组(P0.05),IR+P组上述各指标均明显低于IR组(P0.05)。IR+P组脑组织病理学损伤明显轻于IR组。结论盐酸戊乙奎醚预处理可减轻宫内窘迫致胎鼠脑缺血-再灌注损伤,其作用机制可能与抑制脑组织NF-κB信号通路活性有关。     

盐酸戊乙奎醚对心肺转流下心脏瓣膜置换术患者急性肺损伤的影响

目的探讨盐酸戊乙奎醚(PHC)对心肺转流(CPB)下心脏瓣膜置换术患者术后急性肺损伤(ALI)的影响。方法选择择期行CPB下心脏瓣膜置换术的风湿性心脏病患者30例,男18例,女12例,心功能NYHA分级Ⅱ或Ⅲ级,按照随机数字法分为PHC处理组(P组)和对照组(C组),每组15例。麻醉诱导前P组患者静脉注射PHC 0.03mg/kg,C组患者注射等量生理盐水。分别于麻醉诱导前(T0)、手术结束时(T_1)、术后6h(T_2)、12h(T_3)、24h(T_4)抽取血标本测定血浆IL-6、TNF-α和NF-κB等细胞因子水平;分别于T_0、T_1和T_4时监测动脉血气并计算氧合指数(OI);记录麻醉后即刻和手术结束即刻患者的肺动态顺应性参数。记录患者气管导管留置时间、ICU住院时间和术后住院时间等。结果 P组术后气管导管留置时间、ICU住院时间和术后住院时间均短于C组,但差异无统计学意义。与T_0时比较,T_1~T_4时两组血浆IL-6、TNF-α和NF-κB浓度均明显升高(P0.05);T_1~T_4时P组血浆IL-6、TNF-α和NF-κB浓度均明显低于C组(P0.05)。与T_0和T_1时比较,T4时两组OI明显降低(P0.05);T_4时P组OI高于C组,但差异无统计学意义。与麻醉后即刻比较,手术结束即刻C组肺动态顺应性明显下降(P0.05);手术结束即刻P组肺动态顺应性明显高于C组(P0.05)。结论 PHC能有效抑制CPB下心脏瓣膜手术患者的炎症反应、改善肺顺应性、改善组织氧供,促进患者的术后康复,其机制可能与PHC调控NF-κB信号通路的活性、减少IL-6和TNF-α等促炎细胞因子的释放有关。     

The effects of polymicrobial sepsis with diabetes mellitus on kidney tissues in ovariectomized rats

Objectives: Sepsis model was used to understand the role of sustained hyperglycemia and ovariectomy, either separately or concomitantly, on the response of the activity of the nuclear factor kappa B (NF-κB) and the oxidative response in kidney. Subjects: Polymicrobial sepsis was induced by cecal ligation and puncture (CLP). Diabetes was induced in female rats using administration of alloxan. The rats were divided into five groups: sham control (group 1), ovariectomy (group 2), ovariectomy + sepsis (group 3), ovariectomy + diabetes (group 4), and ovariectomy + diabetic + sepsis (group 5). Results: In kidney tissues, the levels of lipid peroxidation (LPO) and glutathione (GSH) and the activity of catalase (CAT) were higher for groups 3, 4, 5 than the control groups. Superoxide dismutase (SOD) activity was lower for groups 3, 4, 5 than the control groups. We determined that CLP produced injury evident in the kidneys of rats when compared to the control group, whereas the severity of the injury was higher in the diabetes + ovariectomy + CLP group when compared to the CLP group. In immunohistochemical staining, we determined that CLP operation increased NF-κB activation. In the ovariectomized, septic, and diabetic group, NF-κB activation was significantly higher than other groups. Conclusions: Hyperglycemia and ovariectomy severely increased NF-κB activation and oxidant levels with the stages of our sepsis model. Ovariectomy resulted in general changes in metabolism, which are seen in the kidney with diabetes under sepsis conditions.     

Effect and mechanism of soluble epoxide hydrolase inhibitor in renal fibrosis mice model

Objective To investigate the effect and mechanism of soluble epoxide hydrolase inhibitor (sEHI) for NF-κB pathway and cell circle arrest of tubular epithelial cell in unilateral ureteral obstruction (UUO) mice model. Methods Thirty-two healthy C57BL/6 male mice performed UUO surgery to induce renal interstitial fibrosis. Animals were randomly divided into 4 groups: sham group (n=8), sEHI (1 mg?kg-1?d-1) group (n=8), UUO group (n=8) and UUO+sEHI (1 mg?kg-1?d-1) group (n=8). Daily sEHI [1-(1-methylsulfonyl-piperidin-4-yl)-3-(4-trifluoromethoxy-phenyl)-urea, TUPS] or 2% DMSO was applied to mice by oral gavage from day 1 to day 14 after surgery. All mice were sacrificed at day 14 and kidneys were harvested for further analysis. The changes of renal tissue morphology and pathology were observed by Hematoxylin and eosin (HE) and sirius red staining. The expressions of sEH, nuclear factor κB p65 (NF-κB p65) and IκB were measured by Western blotting. The expressions of TNF-α, IL-1β, MCP-1, IL-6, TGF-β, CTGF, collagen-IV and α-SMA were analyzed by real-time PCR. Immunofluorescence staining of phospho-histone H3 (p-HH3) and Ki67 was performed to determine the stage of cell cycle G2/M arrest. Results The expression and activity of sEH increased in UUO group (P﹤0.05). Administration of sEHI inhibited activity of sEH and infiltration of inflammatory cell in tubular interstitial, as well as attenuated tubular damage and tubular interstitial fibrosis. Western blotting analysis revealed administration of sEHI inhibited up-regulated NF-κB p65 and down-regulated IκB in UUO group (P﹤0.05). Real-time PCR demonstrated that administration of sEHI obviously decreased the mRNA expression of cytokines and fibrosis markers, including of TNF-α, IL-1β, MCP-1, IL-6, TGF-β, CTGF, Collagen-IV, α-SMA (P﹤0.05). Immunofluorescence staining showed that there were much more p-HH3 and Ki67 double positive nuclear tubular epithelial cells and interstitial cells in UUO group, compared with Sham group (P﹤0.05). Administration of sEHI reduced the number of double positive nuclear cell only in tubular epithelial cells (P﹤0.05), but not in interstitial cells. Conclusions In UUO tubular interstitial fibrosis model, sEHI inhibits the activation of NF-κB pathway by down-regulating p65 and up-regulating IκB and ameliorates the infiltration of inflammatory cells. In addition, sEHI plays anti-fibrosis effect by moderating cell cycle G2/M arrest and reducing the excrete of pro-fibrosis factors of tubular epithelial cells.