p38抑制剂对缺血再灌注大鼠心肌细胞丝裂原活化蛋白激酶通路的影响

目的探讨p38抑制剂对心肌缺血再灌注大鼠心肌细胞丝裂原活化蛋白激酶(MAPK)通路的影响。方法将44只大鼠随机分为4组:对照组(9只)、缺血组(15只)、缺血再灌注组(10只)和干预组(10只)。对照组大鼠只暴露心脏,缺血组大鼠制备心肌缺血模型,缺血再灌注组大鼠制备心肌缺血再灌注模型,干预组大鼠在左冠状动脉前降支结扎前30 min注射p38抑制剂SB20358(100μg·kg-1),其余操作均与缺血再灌注组一致。观察各组大鼠心肌组织形态学变化及心肌组织中p38、c-Jun氨基端激酶(JNK)和细胞外调节蛋白激酶(ERK1/2)水平的变化。结果对照组大鼠心肌组织中未检测到p38、ERK1/2和JNK蛋白。缺血组缺血60、90 min时大鼠心肌组织中p38蛋白水平显著高于缺血30 min时,差异均有统计学意义(P<0.05);各个时间点,干预组大鼠心肌组织中p38蛋白水平显著低于缺血组和缺血再灌注组,差异均有统计学意义(P<0.05)。随着再灌注时间的延长,缺血再灌注组和干预组大鼠心肌组织中p38蛋白水平逐渐降低,但差异均无统计学意义(P>0.05)。随着缺血和再灌注时间延长,缺血组大鼠心肌组织中ERK1/2和JNK水平略有升高,缺血再灌注组和干预组大鼠心肌组织中ERK1/2和JNK水平略有降低,但差异均无统计学意义(P>0.05)。结论 MAPK中p38参与心肌缺血再灌注损伤,但JNK和ERK1/2的作用不显著。     

Role of p38 mitogen-activated protein kinases in cardioprotection of morphine preconditioning

Background p38 mitogen-activated protein kinases (MAPK) in ischemic preconditioning (IPC) may be essential to cardioprotection. We assessed whether protective effect of morphine-induced preconditioning (MPC) on myocardial ischemia and reperfusion injury in rat hearts involved p38 MAPK activation. Methods Male Spargue-Dawley rats (weighing 300－350 g) were randomly assigned to 1 of the following 8 groups: control (CON, saline vehicle, n=9), SB 203580 (SB, a p38 MAPK inhibitor, n=6), MPC (n=6), IPC (n=9), SB+MPC, SB+IPC, MPC+SB, and IPC+SB (n=6). Infarct sizes (IS), a percentage of the area at risk (AAR), were determined by triphenyltetrazolium (TTC) staining. Tissue samples were processed from the entire AAR of left ventricle for the determination of p38 MAPK protein expression (5 hearts/group). The bands representing the proteins were visualized using an enhanced chemiluminescence detection system. Results The IS/AAR was significantly reduced by IPC (12.9±1.6)% or MPC (25.3±2.9)% compared to the control (52.7±5.5)%. SB 203580 administered prior to preconditioning abolished the effect of IPC (SB+IPC: (43.8±2.6)%, P&gt;0.05 vs CON, P&lt;0.01 vs IPC), but not MPC (SB+MPC: (30.7±0.9)%, P&lt;0.01 vs CON, P&gt;0.05 vs MPC). Treatment with SB 203580 prior to sustained ischemia diminished the protective effect of both MPC (MPC+SB: (42.4±2.9)%, P&gt;0.05 vs CON) and IPC (IPC+SB: (52.0±2.5)%, P&gt;0.05 vs CON) on IS/AAR. In the IPC group, phospho-p38 MAPK protein increased significantly within 5 minutes into ischemia and remained elevated at 30 minutes into reperfusion, while phospho-p38 MAPK protein in the MPC group only increased significantly at 30 minutes into reperfusion. Conclusion The activation of p38 MAPK just acts as a mediator of MPC,whereas it acts as both a trigger and a mediator in IPC.     

通络方剂对糖尿病大鼠肾脏p38 MAPK-FN通路的影响

目的研究通络方剂(TLR)对链脲佐菌素(STZ)诱导的糖尿病大鼠肾脏p38 MAPK-纤维连接蛋白(FN)信号转导通路的影响,探讨TLR对肾脏保护作用的可能机制。方法将SD大鼠随机分成3组(n=7):正常对照组(CON组)、糖尿病未干预组(DM组)和糖尿病TLR干预组(DM+TLR组),DM组和DM+TLR组予以STZ(65 mg/kg)腹腔注射诱导糖尿病模型。制模成功后,DM+TLR组大鼠予以1 g/(kg.d)TLR灌胃,CON组和DM组用等剂量的双蒸水灌胃,持续12周。用自动化生化分析仪检测生化指标,用考马斯亮蓝法定量测定24 h尿蛋白,肾皮质进行PAS染色,用蛋白印迹分析法检测t-p38MAPK、p-p38 MAPK、t-CREB、p-CREB、FN的表达。结果与CON组相比,DM组大鼠的左肾质量/体质量、血糖、尿素氮、血肌酐、24 h尿蛋白、尿蛋白/肌酐增加(P<0.05),体质量减少(P<0.05),p-p38 MAPK、p-CREB、FN表达水平分别增加2.37倍、2.21倍、2.03倍(P<0.05);与DM组相比,DM+TLR组左肾质量/体质量、血肌酐、24 h尿蛋白、尿蛋白/肌酐降低(P<0.05),p-p38 MAPK、p-CREB、FN表达水平降低(P<0.05)。结论 TLR可能通过抑制p38 MAPK-FN通路对糖尿病肾病大鼠的肾脏产生保护作用。     

Immunomodulatory effect of oxymatrine on induced CCl4-hepatic fibrosis in rats

Liver fibrosis is a serious but reversible consequence of liver injuries. The treatment should include clearance of pathogeny, anti-fibrosis and heteropathy. As a potent ingredient extracted from traditional Chinese medicial herbs Sophora flavecientis and S. subprostratae ,oxymatrine contains a variety of alkaloids. Studies have shown that oxymatrine could relieve the inflammatory activity in liver tissue and has the effect of anti-fibrosis as well, whereas the therapeutic mechanism is not clear yet.The present study was designed to investigate the immunological mechanism of oxymatrine therapy for hepatic fibrosis.     

Involvement of the p38 mitogen-activated protein kinase signal transduction pathway in burns-induced lung injury

Acute lung injury (ALl) is a leading complicationin extensively burned patients, especially those with inhalation injury. It can cause hypoxia resulting in injury of remote organs and dysfunction.     

p38蛋白激酶(p38 MAPK)在癫疒间大鼠脑内的表达

 目的  观察p38蛋白激酶(p38 mitogen-activated protein kinase,p38 MAPK)在癫疒间大鼠脑内的表达情况。方法  健康雄性SD大鼠随机分成正常对照组(n=8)和癫疒间组(n=8)。采用戊四氮腹腔注射建立癫疒间模型,大鼠点燃后的惊厥行为按照Racine的标准进行观察评分,采用Western blot和免疫荧光法比较两组大鼠脑内p38 MAPK的表达情况。结果  癫疒间组大鼠脑内p38 MAPK在皮层和海马的表达均显著高于正常对照组(P<0.01)。结论  p38 MAPK在癫疒间大鼠脑内表达上调。     

阿魏酸钠对神经病理痛大鼠痛觉的改善及对p38MAPK信号通路的影响

目的 研究阿魏酸钠对神经病理痛大鼠痛觉的改善作用及对p38MAPK信号通路的影响。方法 建立SD大鼠CCI模型,将40只SPF级健康SD雄性大鼠分为假手术组、模型组、阿魏酸钠组、p38MAPK抑制组,每组10只。观察各组大鼠疼痛行为学、脊髓背角5-HT、GABA浓度及p38MAPK、p-p38MAPK、CREB、p-CREB蛋白变化。结果 阿魏酸钠组及p38MAPK抑制组建模后MWT及脊髓背角5-HT、GABA浓度均低于假手术组而高于模型组,TWL短于假手术组而长于模型组,p-p38MAPK/p38MAPK、p-CREB/CREB均高于假手术组而低于模型组,差异均有统计学意义（P＜0.05）,阿魏酸钠组及p38MAPK抑制组上述指标比较,差异无统计学意义（P＞0.05）。结论 阿魏酸钠能抑制大鼠神经病理痛,其机制可能与抑制p38MAPK信号通路的激活有关。     

p38丝裂原活化蛋白激酶在雄性小鼠生殖细胞株中的表达

目的 检测p38 丝裂原活化蛋白激酶(p38 mitogen-activated protein kinase,p38 MAPK)在雄性小鼠生殖细胞株TM4、GC-1spg、GC-2spd(ts)中的表达,为深入研究p38 MAPK基因在雄性小鼠精子发生中的作用机制奠定理论基础.方法 应用逆转录-多聚合酶链反应(RT-PCR)检测雄性小鼠生殖细胞株TM4、GC-1spg、GC-2spd(ts)中p38 MAPK基因的表达.结果 p38 MAPK基因在TM4、GC-1spg、GC-2spd(ts)细胞株中均为阳性表达.结论 p38 MAPK基因的表达在雄性小鼠精子发生中可能发挥重要作用.     

黄芩素对糖尿病肾病大鼠肾组织p38MAPK表达的影响

目的 探讨黄芩素对糖尿病肾病大鼠肾组织中p38MAPK、核因子-κB(NF-κB) p65表达的干预作用.方法 将30只SD大鼠随机分为模型组、黄芩素干预组及正常对照组,每组10只,黄芩素干预组大鼠予黄芩素300 mg/(kg·d)灌胃,模型组予等体积0.5％羧甲基纤维素钠灌胃.用ELISA法测定24 h尿白蛋白量,免疫组化法检测p38MAPK、p-p38MAPK及NF-κB p65蛋白定位及表达.结果 模型组大鼠24h尿白蛋白量、p38MAPK、p-p38MAPK及NF-κB p65蛋白表达均明显高于正常对照组;与模型组相比,黄芩素干预组的上述指标均降低(均P＜0.05).结论 黄芩素可能通过影响p38MAPK/NF-κB信号通路对糖尿病大鼠肾脏起保护作用.     

银杏苦内酯B对重症急性胰腺炎大鼠p38丝裂原活化蛋白激酶活性的影响

目的观察银杏苦内酯B(BN52021)对重症急性胰腺炎(SAP)大鼠胰腺组织p38丝裂原活化蛋白激酶(p38MAPK)活性的影响。方法Wistar雄性大鼠随机分为阴性对照组(NC组)、SAP模型组(SAP组)、BN52021治疗组(BN组),每组按术后不同时相点(1、2、3、6、12、24h)分为6小组,用Western blotting法分别检测胰腺组织p38MAPK表达与激活情况,同时对胰腺组织进行病理学观察和测定血清淀粉酶活性的变化。结果血清淀粉酶和病理学结果显示SAP造模成功,BN52021能降低SAP大鼠的血清淀粉酶活性,病理学评分在3、6、12h较SAP组显著降低(P<0.05);NC组在各时相点均只检测到少量磷酸化的p38MAPK,SAP组和BN组在各时相点均较NC组显著升高(P<0.05),BN组在6、12、24h较SAP组显著降低(P<0.05)。结论BN52021可部分抑制SAP大鼠胰腺组织p38MAPK的活化,从而发挥其治疗作用。     

Effect of oxymatrine on hepatic gene expression profile in experimental liver fibrosis of rats

Objective:To investigate the effects of oxymatrine on hepatic gene expression profile in a rat model of liver fibrosis.Methods:Forty healthy male SD rats were randomly divided into three groups,a normal group（n=8）,a model group（n=16）,and an oxymatrine treatment group（n=16）.Experimental hepatic fibrosis was induced by subcutaneous injection of carbon tetrachloride（CCI₄）.The rats in the treatment group received oxymatrine via celiac injection at a dosage of 40 mg/kg once a day at the same time.The rats in the model and normal groups received saline at the same dosage via celiac injection.Serum levels of aspartate aminotransferase （AST）,alanine transarninase（ALT）,alkaline phosphatase（AKP）,hyaluronic acid（HA）,and laminin（LN）were assayed.The deposition of collagen was observed with HE and Masson staining.Effect of oxymatrine on hepatic gene expression profile was detected by oligonucleotide microarray analysis with Affymetrix gene chip rat U230A. Quantitative real-time polymerase chain reaction（QRT-PCR）was carried out to confirm the expression changes of six genes.Results:Oxymatrine significantly improved liver function,lowered serum levels of HA and LN,and decreased the degree of liver fibrosis,compared with the model group（P<0.05）.A total of 754 differentially expressed genes were identified by gene chip between the model group and the normal group,among which 438 genes increased and 316 genes decreased over two folds.Compared with the model group,86 genes were downregulated markedly in the oxymatrine group（P<0.05）,including collagen I and other genes related to extracellular material（ECM）,integrin signal transduction genes,early growth response factor genes,and proinflammatory genes;28 genes were upregulated significantly（P<0.05）,including cytochrome P450（CYP450） superfamily genes,glycolipids metabolism and biological transformation related genes.Six genes were confirmed with QRT-PCR,consistent with the result from microarray.Conclusion:Oxymatrine could affect the expression of many functional genes and may be useful in the prevention and treatment of liver fibrosis.     

西尼罗病毒对人神经母细胞瘤细胞p38丝裂原活化蛋白激酶途径的调控

目的 探讨西尼罗病毒（WNV）感染人神经母细胞瘤细胞SH-SY5Y后对p38丝裂原活化蛋白激酶（MAPK）途径的影响,以及该途径在WNV复制及应激应答、炎性应答相关分子表达调控中的作用。方法 WNV分别短时孵育（5、15、30、60 min）和感染（12、24、48、60 h）SH-SY5Y细胞,用蛋白质印迹法检测p38 MAPK和磷酸化p38 MAPK表达,用qRT-PCR检测WNV感染细胞内C/EBP同源蛋白（CHOP）、白细胞介素6（IL-6）、活化转录因子6α（ATF6α）和干扰素刺激基因15（ISG15）mRNA表达水平的动态变化。用WNV感染p38 MAPK siRNA转染的SH-SY5Y细胞,用qRT-PCR检测WNV RNA水平及CHOP、IL-6、ATF6α和ISG15 mRNA水平。结果 与未孵育的细胞相比,WNV短时孵育细胞内p38 MAPK的磷酸化水平升高。WNV感染SH-SY5Y细胞12 h、24 h时激活了p38 MAPK途径,而感染48 h、60 h时明显抑制了p38 MAPK途径。WNV感染促进了CHOP、IL-6和ISG15 mRNA表达而抑制了ATF6α mRNA表达。与对照siRNA转染的细胞相比,p38 MAPK siRNA转染的细胞内WNV RNA（P<0.05）水平和ATF6α mRNA（P<0.01）水平升高,而CHOP mRNA水平降低（P<0.05）。结论 WNV感染早期激活了p38 MAPK途径,该途径的激活可能负反馈调控WNV的复制。WNV经p38 MAPK途径调控与应激应答相关的CHOP、ATF6α表达及与炎性应答相关的IL-6表达。     

p38信号蛋白在乳腺浸润性导管癌中的表达及其与淋巴结转移的关系

目的：探讨p38信号蛋白在乳腺浸润性导管癌中的表达及其与临床病理特征之间的关系。方法：应用免疫组织化学S-P法检测60例乳腺浸润性导管癌，Western blot方法检测30例乳腺浸润性导管癌及5例癌旁正常组织中p38、磷酸化p38（p-p38）的表达，分析p38、p-p38与淋巴结转移等临床病理特征之间的关系。结果：免疫组化结果显示，p38蛋白主要分布于乳腺癌上皮细胞与正常乳腺上皮细胞的胞浆内，癌细胞胞浆浓染，正常乳腺细胞胞浆淡染；p-p38蛋白阳性信号定位于乳腺癌细胞核。有淋巴结转移与无淋巴结转移的乳腺癌组织中p38的阳性率分别为76.3％和40．9％，差异十分显著（P〈0．01）；p-p38的阳性率分别为68.4％和36.4％。差异显著（P〈0．05）。p38、p-p38表达与临床分期有关，与患者年龄、肿瘤大小无明显相关性。Western blot也证实。p38、p-p38在有淋巴结转移患者中表达高于无淋巴结转移者（P〈0．05）。结论：p38、p-p38表达与乳腺癌淋巴结转移有关，p38MAPK信号传导途径在乳腺癌侵袭与转移中起重要作用。     

高氧对幼鼠肺组织细胞凋亡及p38 MAPK丝裂原活化蛋白激酶的影响

目的 探讨高氧对幼鼠肺组织细胞凋亡及p38 MAPK丝裂原活化蛋白激酶表达的影响.方法 幼年Wistar大鼠90%氧气暴露建立高氧肺损伤模型,行肺组织病理学检查,应用TUNEL法检测肺组织的细胞凋亡,Western blot法检测p38 MAPK表达.结果 高氧暴露3 d可见肺组织水肿、出血、炎症细胞浸润等急性肺损伤改变.高氧3 d组的肺凋亡指数较空气对照组明显增加,凋亡发生的主要部位为肺泡、支气管上皮及血管内皮细胞.Westem blot结果显示高氧暴露1 d,肺组织的p38MAPK活性迅速升高,于第2天达到高峰,第3天活性有所下降,但仍高于空气对照组.结论 高氧暴露可以诱导肺组织发生细胞凋亡;高氧可以激活p38MAPK通路,参与高氧性肺损伤的调控;活化的p38MAPK可能参与了高氧诱导的肺组织细胞凋亡的调控.     

鞘内注射氯胺酮对切口痛大鼠脊髓磷酸化p38丝裂原活化蛋白激酶和cAMP反应元件结合蛋白表达的影响

目的观察鞘内注射氯胺酮对切口痛大鼠脊髓化p38丝裂原活化蛋白激酶(p38MAPK)和cAMP反应元件结合蛋白(pCREB)含量的影响。方法成年雄性Sprague-Dawley(SD)大鼠36只,体重250~300 g,随机分为3组:假手术组(Ⅰ组),0.9%氯化钠溶液组(Ⅱ组,鞘内注射0.9%氯化钠溶液25μL),氯胺酮组(Ⅲ组,鞘内注射100μg氯胺酮)。Ⅱ组、Ⅲ组均在切口痛术后立即给药,Ⅲ组鞘内给药用0.9%氯化钠溶液稀释至20μL,注射药物后再用0.9%氯化钠溶液5μL冲洗导管。采用von Frey丝测定术前1 h(T0)及术后2(T1)、4(T2)、8(T3)、16(T4)、24 h(T5)时大鼠后爪机械缩足阈值(PWT值),随后在T5时间点取大鼠脊髓腰膨大部,采用Western印迹法测定磷酸化p38MAPK(p-p38MAPK)和CREB(pCREB)的表达。结果与Ⅱ组比较,Ⅰ组、Ⅲ组在术后各时间点(T1~T5时间点)的PWT值均显著升高(P值均<0.05)。在T5时间点,与Ⅰ组比较,Ⅱ组大鼠脊髓p-p38MAPK和pCREB表达量均显著升高(P值均<0.05);与Ⅱ组比较,Ⅲ组大鼠脊髓p-p38M...     

Ammonia induces upregulation of aquaporin-4 in neocortical astrocytes of rats through the p38 mitogen-activated protein kinase pathway

Background Astrocyte swelling is an important consequence of hepatic encephalopathy, and aquaporin-4 has been reported to play a vital role in this swelling. Ammonia causes astrocyte swelling and is also known to modulate aquaporin-4 expression in the astrocyte foot processes. The purpose of this study was to explore the mechanism of ammonia-induced aquaporin-4 expression, which has been suggested to involve the p38 mitogen-activated protein kinase pathway. Methods We exposed cultured astrocytes to ammonium chloride, an in vitro model of hepatic encephalopathy. The purity of cultured astrocytes was evaluated by fluorescent glial fibrillary acidic protein labeling; cell morphology was assessed by light microscopy; the expression of aquaporin-4, phospho-p38, and p38 were detected by Western blotting analysis. Statistical analysis was performed by one-way factorial analysis of variance, and the relationship between variables was calculated by linear regression using SPSS version 13.0 program for Windows （SPSS, Chicago, IL, USA）. Results The purity of cultured astrocytes was （96.6±1.4）%. Astrocytes swelled significantly when exposed to 5 mmol/L ammonium chloride for 24 hours as compared to non-exposed astrocytes. Co-treatment with 10 μmol/L SB203580 （an inhibitor of p38） attenuated the degree of ammonium chloride induced astrocyte swelling. Western blotting analysis revealed that the expression levels of phospho-p38 and aquaporin-4 in ammonium chloride treated cells were significantly increased relative to the control group （P 〈0.001）; SB203580 co-treatment inhibited the increased expression of phospho-p38 and aquaporin-4 relative to the ammonium chloride treated group （P=0.002 and P=0.015 respectively）. The phosphorylation of p38 and upregulation of aquaporin-4 were highly correlated （r=0.909）. There were no significant differences in total p38 expression among the groups （P=0.341）. Conclusions Ammonium chloride induced upregulation of aquaporin-4 in astrocytes is regulated by the p38 mitogen-activated protein kinase pathway. Inhibiting p38 activation prevented ammonium chloride induced aquaporin-4 protein upregulation.     

氧化苦参碱对大鼠肝纤维化的影响

目的：探讨氧化苦参碱对实验性肝损伤肝胶原合成度及纤维化的影响。方法：采用CCl4造成大鼠慢性肝损伤肝纤维化模型,同时用氧化苦参碱防治,并动态观察血清丙氨酸氨基转移酶（ALT）、Ⅳ型胶原（Ⅳ-C）、透明质酸（HA）、肿瘤坏死因子-α（TNFα）的水平及肝组织病理变化,通过计算机图像分析系统分析肝内纤维组织增生情况。结果：氧化苦参碱治疗组血清ALT,Ⅳ-C,HA,TNFα水平及肝组织内炎症活动度、纤维组织增生程度均低于模型组,且大剂量治疗组又低于小剂量治疗组。结论：氧化苦参碱有减轻肝脏炎症活动度、抑制肝内胶原合成度及抗肝纤维化作用。     

p38丝裂原活化蛋白激酶与骨代谢

丝裂原活化蛋白激酶(mitogen-activated protein kinases,MAPK)是机体广泛表达的丝氨酸/酪氨酸激酶,在哺乳动物细胞多种信号转导通路中起重要作用。p38MAPK信号通路是MAPK通路的一个重要分支,在细胞增殖、分化、凋亡和细胞周期调控等多种生理和病理过程中发挥重要作用。近年来,有关p38MAPK信号通路在与骨代谢相关的破骨细胞、成骨细胞、软骨细胞生长、代谢及功能方面的研究倍受关注。本文就 p38MAPK与骨代谢相关研究进展进行综述,旨在探讨p38MAPK在骨代谢相关疾病中的作用机制。