p38 MAPK抑制剂对脓毒症大鼠肝损伤的保护作用

目的探讨脓毒症肝损害的原因及p38 丝裂原活化蛋白激酶(MAPK)抑制剂的保护作用.方法采用盲肠结扎并穿刺 (CLP)来制作脓毒症模型.在不同时相点观察大鼠肝功能[谷丙转氨酶(ALT)]、血清肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)浓度.结果 CLP术后血清TNF-α、IL-1β进行性升高,ALT也显著升高.血清ALT、TNF-α、IL-1β呈显著正相关.应用p38MAPK抑制剂SB203580后,血清TNF-α、IL-1浓度显著降低,同时血ALT减低.结论 TNF-α、IL-1的大量释放是脓毒症肝损害的原因之一,通过调控p38MAPK信号通路可对脓毒症鼠肝损害起保护作用.     

p38丝裂原活化蛋白激酶抑制剂对脓毒症大鼠多器官损伤的保护作用研究

目的探讨脓毒症致多器官损伤的原因，以及p38丝裂原活化蛋白激酶(MAPK)抑制剂的保护作用和机制。方法采用盲肠结扎穿刺术(CLP)制备脓毒症大鼠模型，治疗组采用术前给予p38MAPK抑制剂SB203580灌胃。在不同时间点观察大鼠血清肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)以及生化指标如丙氨酸转氨酶(ALT)、尿素氮(BUN)、肌酐(Cr)、肌酸磷酸激酶-同工酶(CPK—MB)浓度的变化。结果CLP术后大鼠血清TNF—α、IL-1β显著升高，ALT、BUN、Cr、CPK—MB也进行性升高；血清ALT、BUN、Cr、CPK—MB变化与TNF—α、IL-1β呈显著正相关。应用SB203580后，血清TNF—α、IL-1浓度显著降低，同时ALT、BUN、Cr、CPK—MB也降低。结论TNF—α、IL-1β的大量释放是脓毒症致多器官损伤的原因之一，通过调控p38MAPK信号转导通路可对脓毒症所致多器官损伤起保护作用。     

升降散对脓毒症大鼠心肌p38MAPK蛋白磷酸化水平的影响

目的:探讨升降散保护脓毒症心肌损伤的分子机制。方法:将雄性成年SD大鼠随机分为正常组、模型组、升降散组和p38抑制组。采用盲肠结扎穿孔术(CLP)制备脓毒症大鼠模型,造模后4、8、12、24h进行观察。升降散组和p38抑制组大鼠分别于造模前2h给予升降散灌胃和SB203580皮下注射。留取腹主动脉血标本和心脏组织作相关检测。观察和检测各时间点大鼠死亡率、血清心肌肌钙蛋白I(cTnI)、B型利钠肽(BNP)、白介素-6(IL-6)水平以及心肌p-p38~(MAPK)蛋白、p-p38~(MAPK) mRNA、IL-6mRNA的表达。结果:模型组大鼠24h死亡率25%,升降散组、p38抑制组大鼠死亡率均为15%,组间差异无统计学意义。模型组大鼠血清cTnI、BNP升高;升降散组cTnI造模后8、12、24h较模型组改善(P0.001),BNP造模后4、12、24h较模型组改善(P0.05)。升降散组和p38抑制组cTnI、BNP造模后各时间点差异无统计学意义。模型组大鼠血清IL-6升高,升降散组各时间点IL-6改善(P0.001)。模型组大鼠心肌各时间点p-p38~(MAPK)蛋白水平升高,升降散组各时间点p-p38~(MAPK)蛋白水平均改善(P0.05)。模型组大鼠心肌p-p38~(MAPK) mRNA表达升高,升降散组p-p38~(MAPK) mRNA在8、12、24h改善(P0.05)。模型组大鼠心肌IL-6mRNA升高,升降散组和p38抑制组造模后4、8、12h均改善(P0.05),升降散组IL-6mRNA造模后12h下降优于p38抑制组(P0.05)。结论:升降散可有效降低脓毒症早期大鼠血清cTnI和BNP。升降散改善脓毒症心肌损伤可能与其下调脓毒症早期大鼠心肌p-p38~(MAPK)蛋白水平,降低p-p38~(MAPK) mRNA、IL-6mRNA表达,从而抑制过度炎症反应有关。     

脓毒症小鼠心肌P-选择素基因的表达及意义

目的:探讨脓毒症时小鼠心肌P-选择素表达的改变及其意义.方法:雌性昆明小鼠72只,随机分为对照组(12只)、假手术组(12只)、盲肠结扎穿孔(CLP)组(48只),CLP法制备脓毒症模型,CLP后2、4、8和12 h处死动物分别留取血和心脏组织,采用双抗夹心酶联免疫法(ELISA)检测血清肌钙蛋白Ⅰ(cTnI),测心肌组织髓过氧化物酶(MPO)活性,采用实时荧光定量逆转录多聚酶链反应(RT-PCR)法检测心肌组织P-选择素mRNA表达.结果:CLP组2、4、8和12 h心肌组织P-选择素mRNA表达进行性升高,与对照组和假手术组相比,差异有统计学意义(P＜0.05).而且CLP后心肌组织p-选择素mRNA表达水平与心肌组织MPO活性及血清肌钙蛋白Ⅰ浓度均呈显著正相关.结论:脓毒症时小鼠心肌损伤时,心肌组织P-选择素表达显著升高,可能与心肌组织中性粒细胞浸润及心肌损伤密切相关.     

β1受体阻滞剂对脓毒症大鼠心肌损伤的影响

目的：探讨β1受体阻滞剂对脓毒症大鼠心肌损伤的影响和机制。方法将健康雄性大鼠72只随机分为对照组、脓毒症组、治疗组,每组24只。脓毒症组采用盲肠结扎穿孔法（ CLP）建立脓毒症模型,对照组仅剖腹而不进行CLP。脓毒症组和对照组在关腹后皮下注射生理盐水3 mL／100 g;治疗组除皮下补液外,经尾静脉注射艾司洛尔15 mg／（ kg? h）持续静脉泵入,分别于3、6、12、24 h采集标本,在每个时间点每组大鼠均为6只,观察三组血清肿瘤坏死因子－α（TNF－α）、心肌组织核转录因子－κB p65（NF－κB p65）、心肌肌钙蛋白I（cTnI）的变化。结果与对照组比较,脓毒症组在各时间点血清TNF－α浓度、cTnI浓度及心肌组织NF－κB p65表达均显著升高（P＜0.05）;与脓毒症组比较,治疗组在各时间点血清TNF－α浓度、cTnI浓度及心肌组织NF－κB p65表达均降低（ P＜0.05）。结论β1受体阻滞剂艾司洛尔可减轻脓毒症大鼠心肌损伤,其机制可能与抑制心肌NF－κB p65的表达,降低血清促炎介质的产生有关。β1受体阻滞剂能改善脓毒症失控的炎症反应以及保护心肌功能。     

栀子苷通过调控单核细胞表型改善小鼠盲肠结扎穿孔技术脓毒症模型预后

目的:探讨不同剂量栀子苷治疗脓毒症的疗效和主要生物学机制。方法:雄性BALB/c小鼠通过盲肠结扎穿孔技术(cecal ligation and puncture, CLP)复制脓毒症模型。在生存实验中，动物被随机分为以下各组，每组20只：于CLP术后0 h、24 h经小鼠尾静脉分别注射栀子苷20 mg/kg、40 mg/kg或生理盐水(对照组);于CLP术后24 h经小鼠尾静脉注射栀子苷40 mg/kg或生理盐水(对照组)。观察不同组别的生存预后；并流式检测单核细胞CD16、MHC-Ⅱ、TLR2、TLR4表达水平；ELISA检测血清TNF-α、IL-1β、IL-6、IL-10浓度；Western Blot测定PPARγ浓度。结果:40 mg/kg栀子苷CLP后0 h和24 h静脉给药能显著改善脓毒症小鼠模型生存预后，小剂量(20 mg/kg)栀子苷和延迟给药(24 h)无显著获益。与对照组相比，有效剂量栀子苷能不同程度地全面抑制脓毒症小鼠血清细胞因子TNF-α、IL-1β、IL-6、IL-10浓度，差异有统计学意义(P<0.05);能降低脓毒症小鼠24 h单核细胞CD16表达，差...     

抑制TRAF6调节炎症和自噬改善脓毒症小鼠的心肌损伤和心功能

目的 探索抑制TRAF6后对脓毒症小鼠心肌细胞自噬、炎症反应和心功能的改善作用。方法 （1）昆明雄性小鼠24只,随机分为4组,分别为假手术（sham）组、假手术+C25-140(sham+C)组、盲肠结扎穿刺术（CLP）组、盲肠结扎穿刺术+C25-140(CLP+C)组。sham+C组、CLP+C组术后腹腔注射C25-140。术后24 h超声评估LVEF、LVFS;ELISA测血清TNF-α、IL1-β;HE染色评估心肌炎症反应;透射电镜观察心肌细胞自噬小体、线粒体微结构;qPCR检测心肌组织TRAF6 mRNA;WB检测TRAF6、P62、Beclin-1、LC3B蛋白的表达;（2）予3-MA抑制自噬观察C25-140对脓毒症小鼠心肌损伤的影响。结果 （1）与sham组比,CLP组心肌组织TRAF6mRNA及TRAF6蛋白显著升高（P <0.05）;血清TNF-α、IL1-β浓度显著升高（P <0.05）;心肌组织HE染色见炎症细胞浸润;LVEF、LVFS显著降低（P <0.05）;(2)与CLP组比,CLP+C组TRAF6 mRNA及TRAF6蛋白表达减少（P &...     

不同程度慢性阻塞性肺病患者外周血单个核细胞p38MAPK、NOD2及ROCK1表达及与肺动脉高压、炎症进展的相关性

目的探究不同程度慢性阻塞性肺病(COPD)患者外周血单个核细胞p38丝裂原活化蛋白激酶(p38MAPK)、核苷酸结合寡聚化结构域2(NOD2)及Rho激酶(ROCK1)的表达及其与肺动脉高压和炎症进展的相关性。方法收集60例急性发作期COPD患者(AECOPD)作为AECOPD组,另收集60例COPD稳定期患者,并分为低危组(肺功能分级为Ⅰ～Ⅱ级,n=36)和高危组(肺功能分级为Ⅲ～Ⅳ级,n=24),选择同期体检健康者60例作为健康人对照组。ELISA法测定各组IL-1β、IL-6、IL-8、IL-18、TNF-α、CRP的表达水平;实时荧光定量PCR(RT-PCR)检测各组p38MAPK、NOD2 mRNA的表达水平;流式细胞术检测单个核细胞中ROCK1的表达水平。结果与低危组、高危组和健康人对照组相比,AECOPD组肺动脉平均压、IL-1β、IL-6、IL-8、IL-18、TNF-α及CRP均显著升高,而FEV1、FEV1%及FVC均显著降低(P0.05)。与健康人对照组比较,低危组、高危组及AECOPD组p38MAPK mRNA、NOD2 mRNA及ROCK1的表达水平均显著升高(P0.05)。急性期COPD患者p38MAPK mRNA、NOD2 mRNA及ROCK1表达与FEV1、FEV1%、FVC呈负相关,而与肺动脉平均压、IL-1β、IL-6、IL-8、IL-18、TNF-α及CRP呈正相关(P0.05)。结论 p38MAPK、NOD2及ROCK1参与了COPD患者炎症进展,并与患者肺功能高压、肺功能等病情严重程度存在密切关系。     

p38MAPK和JNK在大鼠肾缺血-再灌注损伤模型中的表达及意义

目的 观察p38MAPK、JNK在肾缺血-再灌注损伤大鼠肾组织中的表达和活化,从而探讨p38MAPK、JNK信号转导通路与肾缺血-再灌注损伤的关系.方法 用缺血1 h再灌注1 h 制备缺血-再灌注模型,20只健康雄性SD大鼠[体质量(200±20)g]随机分为假手术组(n=10)、缺血-再灌注损伤组(n=10).检测肾组织丙二醛(MDA)浓度、超氧化物歧化酶(SOD)活性及血尿素氮(BUN)、肌酐(Cr)浓度.观察肾组织光镜、电镜下的形态学变化,用RT-PCR技术检测两组肾组织p38MAPK、JNK mRNA的表达情况,Western印迹法观察p38MAPK、JNK的活化情况.结果 缺血-再灌注损伤后,大鼠肾功能和肾小管上皮细胞明显受损,BUN、Cr、MDA浓度均高于假手术组(P<0.01),SOD活性低于假手术组(P<0.01),p38MAPK、JNK磷酸化蛋白在假手术组呈少量散在表达或不表达,缺血-再灌注损伤后磷酸化p38MAPK、JNK呈阳性表达(P<0.01).p38MAPK、JNK mRNA在缺血-再灌注损伤组的表达较假手术组显著增高(P<0.01).结论 肾缺血-再灌注可增加p38MAPK、JNK的磷酸化水平及p38MAPK、JNK mRNA的转录水平,p38MAPK、JNK可能在肾缺血-再灌注损伤中起到至关重要的作用.     

p75TNFR激活型单克隆抗体对脑创伤小鼠炎症反应的保护作用

目的 通过单克隆抗体激活p75TNFR信号通路,观察对小鼠脑创伤后关键炎症信号转导通路p38MAPK活化水平和炎症因子表达水平的影响,并探讨其作用机制.方法 参照Feeney等的自由落体法制作小鼠创伤性颅脑损伤模型,运用Western-blot方法检测炎症信号通路中关键分子p38MAPK变化,采用ELISA方法检测注射了D8F2的小鼠血清中炎症因子IL-6、TNF-α和IL-1β水平的变化.结果 创伤后TNF-α、IL-1β、IL-6水平均明显升高,治疗组则低于创伤组,且更快恢复至正常水平.创伤后体内p38MAPK迅速激活至较高水平,治疗组则低于创伤组,且下降更快.结论 在小鼠脑创伤模型中D8F2能通过抑制p38MAPK活化,抑制炎症因子水平的升高,对脑创伤小鼠炎症损害起保护作用.     

P38 MAPK mediates myocardial proinflammatory cytokine production and endotoxin-induced contractile suppression

Cardiac myocytes are capable of synthesizing tumor necrosis factor alpha (TNF-alpha), interleukin-1, and interleukin-6 (IL-1 and IL-6). p38 mitogen-activated protein kinase (MAPK) has been implicated in oxidant-stress-induced myocardial TNF-alpha production; however, the extent to which this kinase contributes to endotoxin-induced contractile dysfunction, as well as TNF-alpha, IL-1alpha, IL-1beta, and IL-6 production, in a bloodless model of endotoxin-induced myocardial dysfunction is unknown. Isolated rat hearts were perfused (Langendorff), and myocardial contractile function continuously recorded, during direct antegrade endotoxin infusion, with and without prior p38 MAPK inhibition. Ventricular p38 MAPK activation (phospho-p38 MAPK Western), cytokine mRNA (RT-PCR), and protein (ELISA) were determined. Endotoxin resulted in progressive decline in left ventricular developed pressure and coronary flow that was attenuated with prior p38 MAPK inhibition (SB 203580). p38 MAPK inhibition significantly decreased endotoxin-induced cardiac TNF-alpha, IL-1alpha, IL-1beta, and IL-6 mRNA levels. To determine the relative effect of TNF-alpha in inducing IL-1alpha, IL-1beta, and IL-6 production, TNF-alpha was sequestered during endotoxin infusion, and TNF-alpha, IL-1beta, and IL-6 protein levels were measured. Interestingly, TNF-alpha sequestration alone significantly decreased myocardial IL-1beta and IL-6 production. We conclude that p38 MAPK is involved in endotoxin-induced myocardial contractile dysfunction and myocardial TNF-alpha production; however, p38 MAPK's involvement in IL-1 and IL-6 production may be indirectly mediated by TNF-alpha.     

Differential effect of caspase inhibition on proinflammatory cytokine release in septic patients

The interleukins (IL)-1beta and IL-18 represent potent players in the proinflammatory cytokine cascade. Their activation is regulated predominantly through the IL-1-converting enzyme (ICE)/caspase-1. The role of caspases in the secretion of IL-1beta and IL-18, as well as in the release of the secondary-induced cytokines IL-12 and interferon (IFN)-gamma in whole blood from septic patients compared to healthy controls, was studied. Inhibition of caspase activity by Z-VAD significantly reduced lipopolysaccharide (LPS) and Staphylococcus aureus (SAC) induced release of mature IL-1beta in septic patients and controls. In contrast, in whole blood from septic patients significantly elevated basal level of IL-18 were found, which could neither be further increased by LPS or SAC, nor be inhibited by Z-VAD. Release of IL-12 p40 was significantly lower in septic patients compared to controls and was not affected by Z-VAD. Despite high levels of IL-18, IFN-gamma was not detected in whole blood from septic patients even after stimulation with SAC or LPS. Thus, during sepsis, caspases participate in the processing of IL-1beta, whereas maturation of IL-18 during sepsis appears to be independent of caspases. The lack of IFN-gamma release seen in septic patients could be attributed to low IL-12 release rather than to diminished IL-18 release.     

Alterations in the cardiac inflammatory response to burn trauma in mice lacking a functional Toll-like receptor 4 gene

Our group and others have previously shown that Toll-like receptor 4 (TLR-4) inactivation prevents burn-induced myocardial contractile dysfunction; however, the molecular mechanisms that are involved in this cardioprotection are not well defined. This present study examines the involvement of TLR-4 in the cardiac inflammatory response to thermal insult. C3H/HeJ (TLR-4 mutant mice) and C3H/HeN wild-type (WT) mice were subjected to either a sham burn or 40% full-thickness burn injury and were fluid resuscitated with lactated Ringer using the Parkland formula. Mice (n = 7-9 per group) were killed at 2, 4, or 24 h postsham or burn, and heart tissue was harvested. Immunoblotting was performed to evaluate phosphorylated p38 mitogen-activated protein kinase (MAPK), nuclear p50, and cytoplasmic p50. Nuclear factor-kappaB was also characterized via electrophoretic mobility shift assay. Systemic and cardiac myocyte secretion of TNF-alpha, IL-1 beta, IL-6, and IL-10 were measured by enzyme-linked immunosorbent assay. Burn injury in WT mice promoted myocardial inflammatory signaling that included increased expression of phosphorylated p38 MAPK, nuclear p50, and increased cardiac myocyte secretion of cytokines. Systemic cytokines were also increased in WT animals, although not to the extent of the myocardial cytokine expression. Toll-like receptor 4 inactivation resulted in an attenuation of several burn-induced responses, including phosphorylation of p38 MAPK, nuclear translocation of nuclear factor-kappaB, and cytokine secretion. These data suggest that burn injury initiates an inflammatory response via Toll/IL-1 signaling in the heart, which contributes to cardiac injury and contractile dysfunction.     

p38MAPK信号通路的变化在心力衰竭病人心肌重塑中的意义

目的：了解不同程度充血性心力衰竭（DHF）病人心肌细胞信号传导通路p38MAPK的变化与心肌重塑和心功能的关系。方法：通过手术取材，采用Western blotting技术测定31例瓣膜病所致CHF病人不同心功能组与5例正常人心肌细胞p38MAPK蛋白基础表达和激活情况。结果：瓣膜病所致心力衰竭病人心肌组织呈心肌重塑的病理改变。心衰组p38MAPK基础表达受抑，激活的磷酸化p38MAPK在轻度心衰者降低，重度心衰病人增高，结论：心衰病人心肌细胞存在MAPK信号传导通路的变化。中，重度心衰病人通过激活p38MAPK诱导心肌细胞凋亡，坏死效应，在心功能恶化中发挥重要病理作用。     

烧伤早期心肌组织几种炎症相关基因表达变化的实验研究

目的 :观察烧伤后心肌组织几种炎症相关基因表达变化 ,探讨其与心肌损害的关系。方法 :采用大鼠4 0 %体表面积 度烫伤模型 ,于伤后 0 h(正常组 )、1h、3h、6 h、12 h、2 4 h用逆转录聚合酶链反应 (RT PCR)方法检测心肌组织肿瘤坏死因子 α(TNFα)、白介素 1β(IL 1β)、诱导型一氧化氮合酶 (i NOS)及胞浆型磷脂酶 A2 (c PL A2 ) m RNA水平 ,四道生理记录仪监测左室收缩压 (L VSP)、左室舒张末压 (L VEDP)和左室压力最大上升 /下降速率 (± dp/dtmax)变化。结果 :烫伤后 1h TNFα和 c PL A2 m RNA表达显著上调 (P均 <0 .0 1) ,此后一直维持高表达状态 ;IL 1β m RNA于伤后 3h表达明显升高 (P<0 .0 1) ,伤后 12 h降至正常水平 ;i NOS m RNA水平除伤后 1h稍上调外 ,其余时间点反而下降。左室收缩功能 (L VSP、+dp/dtmax)和舒张功能 (L VEDP、 dp/dtmax)于伤后 3h显著下降 (P均 <0 .0 1) ,12 h达谷底。左心功能变化与 TNFα、c PL A2表达呈负相关 (P均 <0 .0 5 )。结论 :炎症相关基因 TNFα、c PL A2 及 IL 1β参与了烧伤后心肌局部失控性炎症反应 ,其上调表达可能是烧伤后心肌损害的重要原因之一。     

乌司他丁经p38MAPK通路对脓毒症大鼠急性肾损伤影响的研究

目的研究乌司他丁（UTI）经p38MAPK通路对脓毒症大鼠急性肾损伤（AKI）的影响。 方法采用脂多糖（LPS）诱导脓毒症AKI大鼠模型,将32只SD大鼠随机分为4组,空白对照组、脂多糖组（LPS组）、乌司他丁处理组（LPS+UTI组）、p38MAPK阻断剂干预组（LPS+SB组）,采用酶联免疫吸附试验（ELISA）法检测肾组织中肿瘤坏死因子α（TNF-α）和转化生长因子（TGF-β1）的表达,采用蛋白质免疫印迹试验（Western Blotting）检测肾组织中磷酸化p38MAPK蛋白的表达,并在光镜和电镜下观察肾小球及肾小管的变化。 结果与空白对照组相比,LPS组光镜下肾小球充血肿胀,肾球囊扩张,近曲肾小管上皮细胞肿胀明显,细胞核淡染,部分出现核浓缩、破碎甚至溶解现象,肾间质充血、水肿、大量炎性细胞浸润;电镜下肾小球毛细血管内皮细胞窗孔消失或闭塞,基底膜部分断裂消失,足细胞足突广泛融合消失,近曲肾小管上皮细胞质内线粒体排列紊乱,结构模糊,高度肿胀,有空泡样现象;均提示脓毒症肾损伤严重,且肾组织TNF-α、TGF-β1和p38MAPK蛋白的表达明显升高。与LPS组相比,LPS+UTI组肾组织TNF-α、TGF-β1和p38MAPK蛋白的表达均较LPS组下降［TNF-α（pg/ml）：4915.00±267.06 vs 8836.00±739.51;TGF-β1（pg/ml）：257.71±23.88 vs 354.39±29.44;p-p38MAPK/β-actin：0.158±0.022 vs 0.300±0.044,均P＜0.05］,LPS+SB组肾组织中TNF-α、TGF-β1和p38MAPK蛋白的表达较LPS组均下降［TNF-α（pg/ml）：4856.75±167.23 vs 8836.00±739.51;TGF-β1（pg/ml）：249.56±23.42 vs 354.39±29.44;p-p38MAPK/β-actin：0.136±0.017 vs 0.300±0.044,均P＜0.05］,但LPS+UTI组与LPS+SB组差异无统计学意义（P＞0.05）。 结论乌司他丁对脓毒症急性肾损伤有保护作用,且可能是通过抑制TGF-β1/p38MAPK信号转导通路来实现的。     

Clinical review: Myocardial depression in sepsis and septic shock

Myocardial dysfunction frequently accompanies severe sepsis and septic shock. Whereas myocardial depression was previously considered a preterminal event, it is now clear that cardiac dysfunction as evidenced by biventricular dilatation and reduced ejection fraction is present in most patients with severe sepsis and septic shock. Myocardial depression exists despite a fluid resuscitation-dependent hyperdynamic state that typically persists in septic shock patients until death or recovery. Cardiac function usually recovers within 7-10 days in survivors. Myocardial dysfunction does not appear to be due to myocardial hypoperfusion but due to circulating depressant factors, including the cytokines tumor necrosis factor alpha and IL-1beta. At a cellular level, reduced myocardial contractility seems to be induced by both nitric oxide-dependent and nitric oxide-independent mechanisms. The present paper reviews both the clinical manifestations and the molecular/cellular mechanisms of sepsis-induced myocardial depression.