呼吸机所致肺损伤家兔HSP70和NF-κB表达的意义

目的研究呼吸机所致肺损伤（VILI）炎症反应中热休克蛋白70（HSP70）和核因子-kB（NF-kB）表达的意义，以及加用呼气末正压（PEEP）的影响。方法健康成年新西兰白兔30只，实施麻醉和气管切开术后分别接受不同潮气量（VT）的通气（通气时间均为4h）。随机分为以下三组：PEEP组（VT=40ml/kg，PEEP=3cmH2O,n=12）；ZEEP组（VT=40ml／kg，PEEP=0cmH2O,n=12）；对照组（VT=10ml／kg，n=6），始终以正常条件通气。机械通气开始后4h经颈动脉放血处死动物。在光镜下观察肺组织的病理学改变，Western blot检测家兔肺组织HSP70及NF-kB的表达。结果家兔在致伤机械通气4h后，光镜下可见肺组织有不同程度的损伤；各组肺组织HSP70、NF-kB表达均显著增加，且ZEEP组二者呈负相关。与ZEEP组比较，PEEP组肺组织的病理损伤明显减轻，NF-kB表达明显减少。结论HSP70可通过抑制NF-kB的活性，保护肺组织避免VILI；在机械通气过程中加用PEEP也可通过减少NF-kB的表达减轻肺组织损伤。     

赤芍对大鼠内毒素急性肺损伤丝裂原活化蛋白激酶p38/NF-κB/iNOS的影响

目的 观察赤芍对大鼠内毒素性急性肺损伤(acute lung injury,ALI)丝裂原活化蛋白激酶p38(p38MAPK)/NF-κB/诱导型-氧化氮合酶(iNOS)信号通路的影响,探讨其防治作用的分子机制. 方法采用大鼠内毒素ALI模型,40只Wistar大鼠完全随机分为假手术对照组(A组)、内毒素组(B组)、赤芍治疗组(C组)、赤芍预防组(D组)和SB203580组(E组),每组8只大鼠.观察赤芍对肺组织中p38MAPK、NF-κB和iNOS表达的影响,肺泡灌洗液中性粒细胞比和蛋白含量、肺组织丙二醛(MDA)含量及血清NO含量的变化,于LPS滴注后6 h行血气分析,并进行病理学观察. 结果与A组比较,B、C、D和E组p38MAPK、NF-κB和iNOS表达显著增强,肺泡灌洗液中性粒细胞比和蛋白含量、肺组织MDA含量和血清NO含量显著增加,PaO2和HCO3-明显降低(P<0.05或0.01);与B组比较,C、D组和E组p38MAPK、NF-κB和iNOS表达明显降低(P<0.05);肺泡灌洗液中性粒细胞比和蛋白含量、肺组织MDA显著减少,PaO2和HCO3-明显升高(P<0.05或<0.01).病理学显示C、D组和E组肺组织损伤程度较B组明显减轻;c、D、E组之间各项指标差异无统计学意义.结论 赤芍对内毒素性ALI的防治作用可能与抑制p38MAPK/NF-κB/iNOS信号通路密切相关.     

当归对放射性肺损伤中NF-κB的影响

目的 研究放射性肺损伤中NF-κB活性的变化趋势,并探究当归对其干预及防治肺损伤的作用。方法 实验动物随机分为生理盐水对照组、当归对照组、单纯照射组和当归干预组4组,于1、24、72 h、1、2、4、8、16和24周9个时间点分别取肺组织检测。用HE和Masson染色评价组织病理变化和纤维化;用免疫组化判断NF-κB P65蛋白的定位和相对含量;用TransAM^TM ELISA方法测定NF-κB活性。结果 动物模型的组织病理符合经典改变,4周起出现较明显间质性肺炎,8周最严重, 16周开始肺间质内胶原纤维沉积并渐加重。NF-κB活性在照射后24 h和8周呈现出双相的高峰。预防应用当归后,各时间点当归干预组肺组织的病理改变较同时间点单纯照射组明显要轻,"潜伏期"延长。结论 在放射性肺损伤发展进程中,NF-κB活性呈现双相的升高,预防应用当归能下调这种升高并表现出明显的改善作用。     

当归对小鼠放射性肺损伤中NF-κB的影响

目的 研究小鼠放射性肺损伤中NF-κB活性的变化趋势,并探究当归对其干预及防治肺损伤的作用。方法 实验小鼠分为生理盐水对照组、当归对照组、单纯照射组和当归干预组4组,于1、24、72 h、1、2、4、8、16和24周9个时间点分别取肺组织检测。用HE和Masson染色评价组织病理变化和纤维化;用免疫组化判断NF-κB P65蛋白的定位和相对含量;用TransAM^TM ELISA方法测定NF-κB活性。结果 小鼠模型的组织病理符合经典改变,4周起出现较明显间质性肺炎,8周最严重, 16周开始肺间质内胶原纤维沉积并渐加重。NF-κB活性在照射后24 h和8周呈现出双相的高峰。预防应用当归后,各时间点当归干预组肺组织的病理改变较同时间点单纯照射组明显要轻,"潜伏期"延长。结论 在小鼠放射性肺损伤发展进程中,NF-κB活性呈现双相的升高,预防应用当归能下调这种升高并表现出明显的改善作用。     

乌司他丁对急性肺损伤大鼠肺组织NF-κB表达的影响

目的 研究乌司他丁在脂多糖(LPS)诱导大鼠急性肺损伤(ALD时的肺保护作用机制.方法 36只健康雄性SD大鼠,随机分为3组:正常对照组(NC组),肺损伤组(LPS组)和乌司他丁治疗组(UTI组),每组12只.复制大鼠ALI模型,LPS组经股静脉缓慢注射LPS 5mg/kg;UTI组在注射LPS后,继续由股静脉注入UTI 50 000U/kg;NC组注入2ml 0.9%氯化钠注射液.各组分别于给药完毕后2、4h测定动脉血氧分压(PaO2),然后经颈动脉放血处死6只动物,收集肺标本测定肺组织湿/干(W/D)质量比值,肺组织匀浆中髓过氧化物酶(MPO)活性、丙二醛(MDA)含量,光镜下检查肺组织病理形态学改变,检测肺组织TNF-α蛋白含量和核转录因子-κB(NF-κB)的表达.结果 与NC组比较,LPS组2、4h时的PaO2值均明显下降(P<0.01);UTI组各时间点PaO2均高于LPS组(P<0.05),低于NC组(P<0.05).UTI组和LPS组之间PaCO2未见明显差异(P>0.05).UTI组和LPS组肺组织的W/D比值较NC组均有明显增高(P<0.05),但前两者之间并无明显差异(P>0.05).肺组织病理学检查可见小叶结构紊乱,大量炎性细胞浸润,肺泡腔可见明显出血、渗出,UTI组也存在上述损伤性改变,但程度较LPS组轻.LPS组MPO活性、MDA含量明显高于NC组和UTI组(P<0.05).LPS组TNF-α蛋白含量、NF-κB表达量均明显高于UTI组(P<0.05).结论 在内毒素诱导ALI时,早期使用UTI能够抑制中性粒细胞和NF-κB活化,减少TNF-α的生成,具有较好的肺保护作用.     

抑制NF-κB激活的策略

NF-κB是一个广泛存在于哺乳动物细胞中的转录因子。它可以参与机体的炎症反应、免疫反应、细胞分化与凋亡及其他应激反应。NF-κB启动基因的表达涉及到IKK的激活、IκB磷酸化和泛素化、IκB裂解、NF-κB向核内的迁移、NF-κB与DNA的结合等过程。NF-κB的过度激活会引起一系列的疾病，如哮喘、类风湿性关节炎、肠炎等，利用药物或通过转基因方法来抑制以NF-κB为核心的信号转导途径可能会成为防治某些疾病的途径。     

猪腹部火器伤肠管穿透后NF-κB在心脏中的表达及其意义

目的探讨NF-κB在腹部火器伤肠管穿透后心脏中的变化及意义。方法健康长白仔猪42头随机等分为对照组和伤后1、2、4、8、12 h和24 h组,实验组建立腹部火器伤肠管穿透模型后,用免疫组化图像分析法测定各组心肌组织NF-κB活性,同时测定血清中LDH、CK-MB水平,在光镜下观察各组心脏组织学变化,电镜下观察心脏超微结构改变。结果伤后各组心脏NF-κB表达明显高于对照组。伤后各组血清LDH、CK-MB水平均高于对照组。伤后8、12、24 h组光镜下出现逐渐加重的心肌细胞水肿、变性;电镜下4、8、12、24 h组出现逐渐的线粒体肿胀、溶解;对照组光、电镜下未见明显的损伤性变化。相关分析表明,伤后心肌组织NF-κB表达与血清LDH、CK-MB水平均呈正相关(r值分别为0.968、0.957,P<0.05)。结论腹部火器伤肠管穿透后心肌组织内NF-κB活性增强;NF-κB在腹部火器伤肠管穿透后继发性心脏损伤中具有重要意义。     

海水浸泡对大鼠小肠组织NF-κB,IκBα表达的影响

目的 观察海水浸泡并创伤大鼠小肠NF-κB,IκBα达的变化规律.方法 Wistar大鼠91只,随机分为3组:空白对照组7只,创伤组42只,海水浸泡并创伤组42只.创伤组采用腹部开放伤模型,海水浸泡并创伤组在腹部开放伤的基础上海水浸泡1 h.采用Western blotting方法测定3组小肠组织NF-κB,IκBα达量并进行统计分析和组间比较.结果 与创伤组相比,海水浸泡并创伤组伤后3 h小肠组织NF-κB的表达即出现明显增高,并一直持续到72 h(P<0.05),创伤组似较空白组略有降低,但差异无统计学意义(P>0.05);IκBα变化规律与NF-κB相反.结论 IκBα速而且持续地参与了海水浸泡并创伤大鼠的炎症反应过程,海水浸泡并创伤组大鼠伤情严重,损伤因素持续存在.在相当长一段时间内NF-κB负反馈机制并未得到建立.     

模拟失重对大鼠肺组织NF-κB表达的影响

目的研究模拟失重条件下健康大鼠肺组织中核因子-κB（NF-κB）的表达变化及其意义。方法采用尾悬吊模拟失重大鼠模型,分为悬吊7天、21天组及相应对照组,每组10只健康雄性Wistar大鼠。免疫组织化学法检测肺组织中NF-κB的表达水平。结果悬吊7天及21天组大鼠肺组织NF-κB表达水平与相应对照组比较均显著升高（P〈0．01,P〈0．05）。悬吊21天组大鼠肺组织中NF-κB表达水平较悬吊7天组减弱（P〈0．05）。结论尾悬吊模拟失重大鼠肺组织NF-κB表达水平明显上调,考虑与肺组织损伤有关。     

LPS的直接诱导对肺微血管内皮细胞内NF-κB的影响

目的：探讨LPS的直接诱导作用对肺微血管内皮细胞(PMVEC)核因子κB(NF—κB)的影响。方法：100ng/ml LPS刺激PMVEC 0h、0．5h、1h、2h、4h、6h、8h或10ng/ml、50ng/ml、100ng/ml LPS刺激1h，凝胶电泳迁移率试验检测NF—κB的活化，免疫细胞化学观察其亚基p50、p65的核转位。并通过加入活化阻断剂TPCK观察对诱导的NF—κB活化的最响。结果：LPS的刺激迅速诱导p50、p65亚基核转位，活化NF—κB，1h达到简蜂，且呈剂量依赖关系，后逐渐下降。PDIC能显著抑制其活化，P＜0．01。结论：LPS的直接刺激诱导NF—κB的活化，这可能是LPS诱导炎症反应的一个重要环节。     

NF-κB在放射性损伤中的表达改变及作用机制的研究进展

随着肿瘤发病率的升高,需要接受放射治疗的人数也越来越多;而且由于放射治疗技术的不断发展,患者的长期生存期也在不断延长。因此,在长期生存的患者中防止放射性损伤的发生就尤为重要。NF-κB在各种基因转录的过程中承担着重要作用,并与放射性损伤的发生有着密切关系。笔者就NF-κB的结构、激活、功能作一介绍,并对放射性脑损伤及放射性肺损伤中NF-κB的表达变化、调节机制,以及NF-κB抑制剂的研究进展作一综述。     

CPNE5对NF-κB转录调控活性的影响

目的:研究CPNE5对TNF-α诱导的NF-κB转录激活活性的影响,并探讨其作用机制.方法:用10 ng/mlTNF-α处理转染了pcDNA3.1或pcDNA3.1-CPNE5的HEK293细胞,用双荧光素酶报告基因实验检测不同转染组NF-κB的转录激活活性;Western印迹和细胞免疫荧光技术检测CPNE5表达上调对NF-κB入核的影响;凝胶阻滞实验研究CPNE5表达上调对NF-κB DNA结合能力的影响.结果:CPNE5可以显著抑制TNF-α对NF-κB的转录激活活性(P<0.0001);CPNE5不影响TNF-α诱导的NF-κB(P65)入核;CPEN5过表达降低了TNF-α诱导的NF-κB与DNA结合的能力.结论:CPNE5通过抑制NF-κB的DNA结合活性抑制TNF-α诱导的NF-κB转录激活活性.     

HSP70-2基因多态性与糖尿病肾病易感性的关系

目的 探讨HSP70-2(+1267A/G)基因多态性与2型糖尿病肾病的关系。方法 收集河南省平顶山市152医院肾内科住院及行定期健康体检的人群资料,按照纳入与排除标准纳入研究对象共677例,其中2型糖尿病患者447例(糖尿病肾病组226例,糖尿病非肾病组221例),正常对照组230例。对研究对象进行一般情况问卷调查、体格检查及实验室生化指标检测,并采用PCR-RFLP技术分析HSP70-2基因(+1267A/G)多态性。结果 糖尿病肾病组的HSP70-2基因G/G基因型频率高于糖尿病非肾病组和正常对照组,差异有统计学意义(χ2=8.123,P<0.01;χ2=11.651,P<0.01);糖尿病肾病组的G等位基因频率高于糖尿病非肾病组和正常对照组,差异有统计学意义(χ2=9.392,P<0.01,OR=1.782;χ2=11.971,P<0.01,OR=2.153)。G/G基因型携带者MALB/Cr和BMI水平高于A/A基因携带者,差异有统计学意义(P<0.05)。结论 HSP70-2基因1267位点G/G基因型可能与糖尿病肾病及全身性肥胖易感性相关;G等位基因增加了携带者患糖尿病肾病的风险。     

创伤性急性肺损伤后高迁移率族蛋白B1的临床相关性研究

目的 研究创伤性急性肺损伤(acute lung injury,ALI)后血清高迁移率蛋白-1(high mobility group box 1 protein,HMGB -1)的水平变化,探讨其与MODS、急性生理学与慢性健康状况Ⅱ(acute physiology and chronic health evaluation,APACHEⅡ)评分的相关性,以及其对MODS发生率和死亡率的预警作用. 方法 测定10例正常人与40例ALI患者伤后第1,4,7天的血清HMGB -1水平.根据MODS的诊断标准,将损伤组分为两组:MODS组13例,非MODS组27例.同时评定其MODS、APACHEⅡ分值. 结果 组间比较:非MODS组第1,4,7天HMGB -1水平的表达均高于MODS组(P＜0.05);非MODS组HMGB -1水平的表达高于正常对照组,但低于MODS组.组内比较:非MODS组第4天HMGB -1水平的表达明显高于第1天(P＜0.01),第7天较第4天明显降低(P＜0.01),甚至低于第1天.MODS组血清HMGB -1水平的表达显著升高,并延续数天,第7天较第4天有所降低,但仍高于第1天.伤后第1,4,7天MODS评分随HMGB -1水平的动态变化差异明显.伤后第1,4,7天APACHEⅡ评分比较,差异明显.相关性分析显示,HMGB -1水平的表达与MODS评分、APACHEⅡ评分显著相关. 结论 (1)HMGB -1在ALI患者中呈高表达,其作为晚期炎症介质参与炎性反应往往升高较晚,且持续时间较长.(2)HMGB -1水平变化与并发MODS密切相关.(3)常规检测血清HMGB -1水平并联合评定MODS、APACHEⅡ评分有助于对创伤后脏器功能不全的预测.     

树突状细胞表面HSP70受体介导的内化作用

目的 研究树突状细胞（DCs）表面HsP70受体介导的内化作用。方法 实验分为3组：HsI）70受体封闭组、非受体封闭组和对照组,采用流式细胞仪检测C57BL／6小鼠骨髓来源DCs内化FITC标记的HSP70阳性细胞率。结果 受体封闭组50越HSP70可使DCs表面受体饱和;受体饱和后随HSP70-FITC用量的升高,HSP70-FITC阳性细胞数呈正比性增高;非受体封闭组应用50心HSP70-FITC与100μg HsP70-FITC时HSP70-FITC阳性细胞率均达到90％以上且组间差别不大。结论：DCs表面HSP70受体介导的内化作用是DCs摄取抗原的主要途径,且具有饱和性和优先性的特点。当受体饱和后,非受体介导的内化作用可能在DCs内化HsP70过程中起作用并且呈剂量依赖性。     

NF-κB不同二聚体的活化与辐射诱导凋亡的关系

转录因子κ基因结合核因子(NF-κB)一直被看作是细胞的存活因子,对辐射诱导的细胞凋亡起强烈的抵抗作用。然而,近来的研究逐渐表明,NF-κB不同亚基的组合可能有着不同、甚至完全相反的作用。对这方面课题的深入研究,无疑对理解中、高剂量辐射免疫抑制效应的机制可提供有意义的理论依据,并对临床肿瘤的放疗将提供新的有价值的资料。     

Effects of seawater immersion combined with open abdominal injury on the expression of NF-κB,and IκBαof small intestine in rats

Objective To observe effects of seawater immersion combined with open abdominal injury on the expression of NF-κB,and IκBαas well as the change pattern in rats. Methods Ninety-one Wistar rats were randomly divided into 3 groups: the control group (n =7), the open abdominal injury group(n =42) and open abdominal injury combined with 1-hour seawater immersion group ( n =42). The expression of NF-κB,andIκBαin small intestine tissues was measured by Western blot and statistical analyses were also made in the study. Results The expression of NF-κB,in the seawater immersion combined with open abdominal injury group increased significantly 3 hours after injury, when compared with that of the open abdominal injury group(P<0. 05), whereas the expression of NF-κB, of the pure injury group was slightly lower than that of the control group, but no statistical differences could be seen between them(P>0.05). The change pattern in the expression of IκBαwas quite the opposite to that of NF-κB. Conclusions NF-κB seemed to be rapidly and persistently involved in the whole inflammatory response to trauma induced by opened abdominal injury and seawater immersion, when a comparison was made with the pure open abdominal injury group. Injuries for the rats in the open abdominal injury combined with seawater immersion group were serious, and the feedback mechanism for NF-κB was not established for quite a long time.     

Effects of seawater immersion combined with open abdominal injury on the expression of NF-κB,and IκBαof small intestine in rats

Objective To observe effects of seawater immersion combined with open abdominal injury on the expression of NF-κB,and IκBαas well as the change pattern in rats. Methods Ninety-one Wistar rats were randomly divided into 3 groups: the control group (n =7), the open abdominal injury group(n =42) and open abdominal injury combined with 1-hour seawater immersion group ( n =42). The expression of NF-κB,andIκBαin small intestine tissues was measured by Western blot and statistical analyses were also made in the study. Results The expression of NF-κB,in the seawater immersion combined with open abdominal injury group increased significantly 3 hours after injury, when compared with that of the open abdominal injury group(P<0. 05), whereas the expression of NF-κB, of the pure injury group was slightly lower than that of the control group, but no statistical differences could be seen between them(P>0.05). The change pattern in the expression of IκBαwas quite the opposite to that of NF-κB. Conclusions NF-κB seemed to be rapidly and persistently involved in the whole inflammatory response to trauma induced by opened abdominal injury and seawater immersion, when a comparison was made with the pure open abdominal injury group. Injuries for the rats in the open abdominal injury combined with seawater immersion group were serious, and the feedback mechanism for NF-κB was not established for quite a long time.     

Effects of seawater immersion combined with open abdominal injury on the expression of NF-κB,and IκBαof small intestine in rats

Objective To observe effects of seawater immersion combined with open abdominal injury on the expression of NF-κB,and IκBαas well as the change pattern in rats. Methods Ninety-one Wistar rats were randomly divided into 3 groups: the control group (n =7), the open abdominal injury group(n =42) and open abdominal injury combined with 1-hour seawater immersion group ( n =42). The expression of NF-κB,andIκBαin small intestine tissues was measured by Western blot and statistical analyses were also made in the study. Results The expression of NF-κB,in the seawater immersion combined with open abdominal injury group increased significantly 3 hours after injury, when compared with that of the open abdominal injury group(P<0. 05), whereas the expression of NF-κB, of the pure injury group was slightly lower than that of the control group, but no statistical differences could be seen between them(P>0.05). The change pattern in the expression of IκBαwas quite the opposite to that of NF-κB. Conclusions NF-κB seemed to be rapidly and persistently involved in the whole inflammatory response to trauma induced by opened abdominal injury and seawater immersion, when a comparison was made with the pure open abdominal injury group. Injuries for the rats in the open abdominal injury combined with seawater immersion group were serious, and the feedback mechanism for NF-κB was not established for quite a long time.     

Effects of seawater immersion combined with open abdominal injury on the expression of NF-κB,and IκBαof small intestine in rats

Objective To observe effects of seawater immersion combined with open abdominal injury on the expression of NF-κB,and IκBαas well as the change pattern in rats. Methods Ninety-one Wistar rats were randomly divided into 3 groups: the control group (n =7), the open abdominal injury group(n =42) and open abdominal injury combined with 1-hour seawater immersion group ( n =42). The expression of NF-κB,andIκBαin small intestine tissues was measured by Western blot and statistical analyses were also made in the study. Results The expression of NF-κB,in the seawater immersion combined with open abdominal injury group increased significantly 3 hours after injury, when compared with that of the open abdominal injury group(P<0. 05), whereas the expression of NF-κB, of the pure injury group was slightly lower than that of the control group, but no statistical differences could be seen between them(P>0.05). The change pattern in the expression of IκBαwas quite the opposite to that of NF-κB. Conclusions NF-κB seemed to be rapidly and persistently involved in the whole inflammatory response to trauma induced by opened abdominal injury and seawater immersion, when a comparison was made with the pure open abdominal injury group. Injuries for the rats in the open abdominal injury combined with seawater immersion group were serious, and the feedback mechanism for NF-κB was not established for quite a long time.