GLUT4、TNF-α- mRNA、iNOS在冠心病患者冬眠心肌中的表达和意义

目的探讨冬眠心肌细胞内GLUT4、TNF-α-mRNA、iNOS的变化和意义。方法10例行冠脉搭桥术的冠心病患者,术前一周内用多巴酚丁胺超声负荷试验结合多普勒组织成像确定冬眠心肌及正常心肌的位置,搭桥术中根据检测结果进行取材(分别取正常心肌和冬眠心肌),并经电镜证实。Western-blot法测GLUT4、iNOS蛋白表达情况,ISH法测TNF-α-mRNA的活性,Tunel法测心肌细胞凋亡情况。结果冬眠心肌细胞内GLUT4、TNF-α-mRNA、iNOS的表达及心肌细胞凋亡数较正常细胞高;冬眠心肌TNF-α-mRNA与心肌细胞凋亡数、iNOS的蛋白表达相关(P<0.05),r分别为0.857、0.784。结论慢性缺血缺氧时,心肌细胞内GLUT4、TNF-α-mRNA、iNOS表达增加。TNF-α可介导细胞凋亡,并促进iNOS表达,GLUT4、TNF-α、iNOS是冬眠心肌形成的重要分子生物学因子。     

p38MAPK表达在冠心病患者冬眠心肌细胞凋亡的意义

目的:探讨冠心病患者冬眠心肌细胞内活化的丝裂素活化蛋白激酶(p38MAPK)对心肌细胞凋亡的影响.方法:行冠状动脉搭桥术(CABG)的冠心病患者10例,术前1周内用多巴酚丁胺超声负荷试验结合多普勒组织成像确定冬眠心肌及正常心肌的存在部位,CABG术中根据检测结果进行取材(分别取正常心肌和冬眠心肌),并经电镜证实.取材心肌用Tunel法检测心肌细胞凋亡情况,免疫印迹法(Western-blot)检测磷酸化的p38的表达情况.结果:冬眠心肌细胞内磷酸化p38、心肌细胞凋亡数较正常心肌高;p38与心肌细胞凋亡数相关(P＜0.05,r=0.816).结论:心肌慢性缺血时,心肌细胞内p38MAPK信号活化,活化的p38MAPK介导冬眠心肌细胞凋亡.     

p38丝裂素活化蛋白激酶对冠心病患者冬眠心肌细胞凋亡的影响

目的探讨冬眠心肌细胞内活化的p38丝裂素活化蛋白激酶对冬眠心肌细胞凋亡的影响。方法行冠状动脉搭桥术的冠心病患者10例,术前一周内用多巴酚丁胺超声负荷试验结合多普勒组织成像确定冬眠心肌及正常心肌的存在部位,冠状动脉搭桥术中根据检测结果进行取材(分别取正常心肌和冬眠心肌),并经电镜证实。取材心肌用Tunel法检测心肌细胞凋亡情况,免疫印迹法检测磷酸化p38的表达情况。分析冬眠心肌与正常心肌磷酸化p38及心肌细胞凋亡数是否存在差异;分析p38与心肌细胞凋亡之间的相关性。结果冬眠心肌细胞内磷酸化p38、心肌细胞凋亡数较正常心肌高;p38与心肌细胞凋亡数相关(r=0.816,P<0.05)。结论心肌慢性缺血缺氧时,心肌细胞内p38丝裂素活化蛋白激酶信号活化,活化的p38丝裂素活化蛋白激酶介导冬眠心肌细胞凋亡。     

ERK1/2蛋白在胆管癌组织中的表达及意义

采用SP免疫组化方法检测70例胆管癌组织中p-ERK1/2蛋白的表达.结果 胆管癌组织中p-ERK1/2蛋白阳性表达率为48.6%,p-ERK1/2蛋白表达与病理分化程度显著相关(r=0.294,P<0.05);Kaplan-Meier法生存分析结果显示,p-ERK1/2蛋白的表达与术后无复发生存时间呈负相关(χ2 =11.06,P<0.01).认为胆管癌组织中p-ERK1/2的表达可能和肿瘤的发展有关,对判定预后有一定价值.     

Plk1与Survivin蛋白在肝细胞癌中的表达及意义

目的探讨Plk1和凋亡抑制蛋白Survivin在HCC中表达情况,为临床诊断和治疗提供理论依据。方法采用免疫组化法检测100例肝细胞癌标本中Plk1、Survivin的表达。结果⑴肝细胞癌中,Survivin蛋白的表达明显高于癌旁正常组织（P〈0.05）,Survivin在肝细胞癌组织中的表达与患者性别、年龄、肿瘤分化程度、AFP表达及是否侵犯肝被膜均无关联（P〉0.05）。与HBsAg表达有关联（P〈0.05）。⑵肝细胞癌组织中,Plk1阳性表达明显高于癌旁正常组织（P〈0.05）;Plk1在肝细胞癌组织中的表达与患者性别、年龄、AFP表达、HBsAg表达及是否侵犯肝被膜均无关（P〉0.05）,与肿瘤分化程度相关（P〈0.05）。⑶肝细胞癌组织中,Plk1与Survivin表达呈正相关（P〈0.05）。结论 Plk1与Survivin的联合检测为肝细胞癌的诊断和治疗提供新的依据。     

高血压脑出血患者外周血单个核细胞中P38MAPK的表达

目的探讨P38 MAPK在高血压脑出血外周血单个核细胞中的表达情况及其临床意义。方法选择宁夏医科大学总医院神经外科确诊的高血压脑出血患者50例作为实验组,在发病后1、3、7、14 d分别抽取静脉血,用PCR荧光检测法测定P38 MAPK mRNA的表达,同时ELISA检测外周血IL-6/8含量,与正常体检者(25例,对照组)对比,并分析表达意义。结果实验组患者外周血单个核细胞核内P38 MAPK mRNA表达和血清IL-6/8含量均高于正常体检者(P0.05),病情越重,P38 MAPK mRNA及IL-6/8含量越高,在脑出血后3 d时P38 MAPK mRNA及IL-6/8含量达到高峰,此后表现为下降趋势,P38MAPK mRNA及IL-6/8含量之间呈正相关(P0.05)。结论 P38 MAPK、IL-6、IL-8参与了高血压脑出血的炎症反应,与脑出血后脑组织损伤机制相关。     

磷酸化p38丝裂原活化蛋白激酶在急性肝衰竭小鼠模型及HBV相关慢加急性肝衰竭患者肝组织中的表达及意义

目的研究磷酸化后的p38丝裂原活化蛋白激酶(p-p38MAPK)在氨基半乳糖(D-Gal N)或脂多糖(LPS)诱导的急性肝衰竭小鼠模型以及HBV相关慢加急性肝衰竭(ACLF)患者肝脏中的表达及其意义。方法采用D-Gal N/LPS诱导C57BL/6小鼠构建急性肝衰竭模型,分别在给药0、0.5、1、2、4、6、8 h设立实验组,每组4只,处死小鼠取肝组织标本进行HE染色观察肝组织结构的病理变化,分别运用蛋白免疫印迹技术半定量检测及免疫组化染色定位检测肝组织中p-p38MAPK的表达;同时对照研究pp38MAPK在HBV-ACLF、乙型肝炎肝硬化、慢性乙型肝炎(CHB)患者肝组织中的表达情况。组间比较采用独立样本t检验。结果蛋白免疫印迹法检测p-p38MAPK在肝衰竭小鼠肝组织匀浆中的表达随时间变化持续增高,给药6 h组半定量分析表达量显著高于正常对照组,差异具有统计学意义(t=-2.727,P=0.034)。免疫组化染色结果显示随存活时间的延长,小鼠肝组织炎症程度逐渐加重,炎症早期主要为窦细胞表达p-p38MAPK,随着肝组织损害加重,肝细胞表达p-p38MAPK渐多,坏死肝组织附近分布大量p-p38MAPK阳性肝细胞;正常人肝组织中p-p38MAPK的表达量很低,CHB患者肝组织内可见浸润淋巴细胞及肝细胞表达pp38MAPK,并且随病程进展呈增多趋势,与临床观察病变程度逐渐加重相一致。结论 D-Gal N/LPS诱导的小鼠急性肝衰竭模型中,p-p38MAPK表达随肝组织损害加重,表明其在肝衰竭病变过程中占重要地位;在HBV-ACLF患者致病过程中,p-p38MAPK信号通路可能发挥重要作用。     

溶血磷脂酸、hs-CRP在高胆固醇血症合并冠心病患者中的表达及意义

目的探讨溶血磷脂酸（LPA）、高敏C反应蛋白（hs-CRP）在高胆固醇血症合并冠心病患者中的表达及意义。方法选择100例高胆固醇血症患者,根据ACC/AHA指南关于冠心病的诊断标准,将患者分为急性心肌梗死（AMI）组25例、不稳定型心绞痛（UAP）组25例、稳定型心绞痛（SAP）组25例,单纯高胆固醇血症（对照）组25例。分别采用循环酶法检测LPA、免疫比浊法检测hs-CRP。结果 AMI组、UAP组、SAP组LPA水平明显高于对照组（P均〈0.01）,AMI组、UAP组LPA水平明显高于SAP组（P均〈0.01）;AMI组、UAP组hs-CRP水平明显高于SAP组及对照组（P均〈0.01）;LPA的表达水平与hs-CRP、LDL-C呈正相关（r分别为0.689、0.802,P均〈0.01）。结论 LPA、hs-CRP在高胆固醇血症合并冠心病患者中表达升高,两者联合检测对高胆固醇血症患者预测冠心病的发生及判断预后具有重要的临床价值。     

血清超敏C-反应蛋白、视黄醇结合蛋白4及血浆结合珠蛋白水平在冠心病患者中的变化及其意义

目的探讨冠心病(CHD)患者血清超敏C-反应蛋白(hs-CRP)、视黄醇结合蛋白4(RBP4)及血浆结合珠蛋白(Hp)水平的变化及其临床意义。方法选取2014年2月至2017年2月广安市人民医院心血管内科经冠状动脉造影检查确诊的CHD患者140例作为CHD组,另选取同期在我院心血管内科住院的非CHD患者60例作为对照组。将CHD组患者根据临床表现分为3个亚组:急性心肌梗死(AMI)组(n=38)、不稳定型心绞痛(UAP)组(n=62)和稳定型心绞痛(SAP)组(n=40)。依据病变支数分为3个亚组:单支病变组(n=27)、双支病变组(n=54)和三支病变组(n=59)。检测各组患者的血清hs-CRP、RBP4、Gensini评分及血浆Hp水平,并分析各指标与病情严重程度的关系。采用SPSS 16.0软件进行数据处理。依据数据类型,组间比较分别采用t检验和x~2检验。相关性分析采用Pearson线性相关分析法。结果 CHD组患者的血清hs-CRP[(7.29±3.65)vs(2.84±1.16)mg/L]、RBP4[(49.64±8.32)vs(42.07±5.51)ng/ml]及血浆Hp[(95.26±19.14)vs(70.13±16.33)mg/L]水平均显著高于对照组(P0.05)。AMI亚组患者的血清hs-CRP、RBP4、Gensini评分及血浆Hp水平均显著高于SAP亚组和UAP亚组患者(P0.05);UAP亚组患者的血清hs-CRP、RBP4、Gensini评分及血浆Hp水平均显著高于SAP亚组患者(P0.05)。三支病变亚组患者的血清hs-CRP、RBP4、Gensini评分及血浆Hp水平均显著高于双支病变和单支比病变亚组患者(P0.05);双支病变亚组患者的血清hs-CRP、RBP4、Gensini评分及血浆Hp水平均显著高于单支病变亚组患者(P0.05)。CHD患者的Gensini评分与血清hs-CRP(r=0.619)、RBP4(r=0.554)及血浆Hp(r=0.592)水平均呈显著正相关(P0.05)。结论 CHD患者血清hs-CRP、RBP4及血浆Hp水平显著升高,且与冠状动脉病变严重程度密切相关。     

Prevalence of hibernating myocardium in patients with severely impaired ischaemic left ventricles

OBJECTIVE: Severe impairment of left ventricular (LV) contraction is associated with an adverse prognosis in patients with ischaemic heart disease. Revascularisation may improve the impaired LV contraction if hibernating myocardium is present. The proportion of patients likely to benefit from this intervention is unknown. Therefore, the prevalence of hibernating myocardium in patients with ischaemic heart disease and severe impairment of LV contraction was assessed. DESIGN: From a consecutive series of patients undergoing coronary angiography for the investigation of chest pain or LV impairment, all patients with ischaemic heart disease and an LV ejection fraction (LVEF) < or = 30% were identified. These patients underwent positron emission tomography (PET) to detect hibernating myocardium, identified by perfusion metabolism mismatch. SETTING: A teaching hospital directly serving 500,000 people. RESULTS: Of a total of 301 patients, 36 had ischaemic heart disease and an LVEF < or = 30%. Twenty-seven patients had PET images, while nine patients were not imaged because of emergency revascularisation (three), loss to follow up (one), inability to give consent (four), and age < 50 years (one, ethics committee guidelines). Imaged and non-imaged groups were similar in LV impairment, demographic characteristics, and risk factor profile. Fourteen patients (52% of the imaged or 39% of all patients with ischaemic heart disease and LVEF < or = 30%) had significant areas of hibernating myocardium on PET. CONCLUSION: It is possible that up to 50% of patients with ischaemic heart disease and severely impaired left ventricles have hibernating myocardium.     

Cardiac expression and subcellular localization of the p38 mitogen-activated protein kinase member, stress-activated protein kinase-3 (SAPK3)

Despite the interest in the roles that mitogen-activated protein kinases (MAPKs) play in the heart, the role of the different MAPK isoforms has been relatively poorly defined. A third isoform of p38 MAPK, known variously as stress-activated protein kinase-3 (SAPK3), p38- gamma or ERK6, has been previously shown to differ from p38- alpha/ beta both in its molecular weight and its lack of inhibition by the compound SB203580. We have generated monoclonal antibodies with specificity for SAPK3 demonstrated by immunoblot analysis, immunofluorescence studies, and cloning of SAPK3 from a rat heart cDNA expression library. By immunoblotting, we confirmed high expression of SAPK3 in fast, slow and mixed fibre types of murine skeletal muscle and observed significant expression restricted to heart, lung, thymus and testes. In addition to expression in normal heart (human, mouse, rat, dog and pig), we observed constant expression in diseased human heart, as well as control and hypertrophic cultured neonatal rat cardiac myocytes. Immunolocalization in cultured cardiac myocytes followed by confocal microscopy showed punctate, non-nuclear SAPK3 staining. In contrast, p38- alpha/ beta staining was non-punctate and distributed throughout the cytosol and nucleus. Whereas treatment with Leptomycin B to prevent nuclear export processes promoted higher levels of p38- alpha/ beta staining in cardiac myocyte nuclei, there was no apparent change in SAPK3 localization under these conditions. These differences between p38- alpha/ beta and SAPK3 probably reflect the specialized functions of SAPK3 and emphasize the need to evaluate SAPK3 upstream activators and downstream targets in the heart.     

睡眠剥夺对大鼠颞下颌关节髁突组织p38信号通路相关蛋白表达的影响

目的 探讨睡眠剥夺对大鼠颞下颌关节（TMJ）的破坏作用及对p38信号通路相关蛋白表达的影响.方法 将40只Wistar雄性大鼠随机分为SD组和对照组.SD组采用改良多平台法建立睡眠剥夺模型.于制模4d后处死两组大鼠,HE染色观察TMJ髁突组织病理学变化,免疫组化SP法检测TMJ髁突组织p38信号通路相关蛋白MKK6、p38表达.结果 SD组TMJ髁突表面可见部分胶原纤维水肿、松解,出现炎症反应;SD组MKK6、p38阳性细胞率均高于对照组;MKK6、p38蛋白表达阳性率均高于对照组（P＜0.05）.结论 大鼠睡眠剥夺时TMJ出现病理性改变,p38信号通路可能参与了TMJ关节破坏的病理过程.     

p38和CARP的表达与大鼠心肌梗死后心肌重塑的关系

目的探讨p38丝裂原激活的蛋白激酶（p38）和心锚重复蛋白（CARP）的表达与大鼠心肌梗死（MI）后心肌重塑的关系。方法结扎大鼠冠脉前降支致MI,术后2 h存活大鼠随机分为MI组、p38抑制剂SB203580组（SB组）和假手术组（Sham组）。测定各组第1、7和28天左心室质量指数（LVWI）、心肌细胞横切面面积（CSA）。RT-PCR法测定p38和CARP在心脏的表达。结果与Sham组相比,MI组LVWI和CSA在第7、28天时明显增加（P均〈0.05）,各时间点磷酸化p38（p-p38）表达均明显升高（P均〈0.05）,CARP在第1、7天时表达明显升高（P均〈0.05）。与MI组相比,SB组LVWI在第7天时明显降低（P〈0.01）;CSA在第1天时增大（P〈0.01）,第7、28天时明显缩小（P均〈0.01）;p-p38在第1天时表达降低,CARP在第7天表达降低（P〈0.01）。结论MI后早期p38和CARP即被激活,并促进随后心肌重塑的发展,抑制p38可以抑制CARP,改善心肌重塑。     

磷酸化p38丝裂原活化蛋白激酶在急性肝衰竭小鼠模型及HBV相关慢加急性肝衰竭患者肝组织中的表达及意义磷酸化p38丝裂原活化蛋白激酶在急性肝衰竭小鼠模型及HBV相关慢加急性肝衰竭患者肝组织中的表达及意义

目的：研究磷酸化后的p38丝裂原活化蛋白激酶（p－p38MAPK）在氨基半乳糖（D－GalN）或脂多糖（LPS）诱导的急性肝衰竭小鼠模型以及HBV相关慢加急性肝衰竭（ACLF）患者肝脏中的表达及其意义。方法采用D－GalN／LPS诱导C57BL／6小鼠构建急性肝衰竭模型,分别在给药0、0．5、1、2、4、6、8 h设立实验组,每组4只,处死小鼠取肝组织标本进行HE染色观察肝组织结构的病理变化,分别运用蛋白免疫印迹技术半定量检测及免疫组化染色定位检测肝组织中p－p38MAPK的表达;同时对照研究p－p38MAPK在HBV－ACLF、乙型肝炎肝硬化、慢性乙型肝炎（CHB）患者肝组织中的表达情况。组间比较采用独立样本t检验。结果蛋白免疫印迹法检测p－p38MAPK在肝衰竭小鼠肝组织匀浆中的表达随时间变化持续增高,给药6 h组半定量分析表达量显著高于正常对照组,差异具有统计学意义（t＝－2.727,P＝0．034）。免疫组化染色结果显示随存活时间的延长,小鼠肝组织炎症程度逐渐加重,炎症早期主要为窦细胞表达p－p38MAPK,随着肝组织损害加重,肝细胞表达p－p38MAPK渐多,坏死肝组织附近分布大量p－p38MAPK阳性肝细胞;正常人肝组织中p－p38MAPK的表达量很低,CHB患者肝组织内可见浸润淋巴细胞及肝细胞表达p－p38MAPK,并且随病程进展呈增多趋势,与临床观察病变程度逐渐加重相一致。结论 D－GalN／LPS诱导的小鼠急性肝衰竭模型中,p－p38MAPK表达随肝组织损害加重,表明其在肝衰竭病变过程中占重要地位;在HBV－ACLF患者致病过程中,p－p38MAPK信号通路可能发挥重要作用。     

Hibernating myocardium in patients with coronary artery disease: Identification and clinical importance

Summary The term hibernating myocardium describes a particular outcome of myocardial ischemia in which myocytes show a chronically depressed contractile ability but remain viable. Revascularization of hibernating tissue causes a recovery of mechanical function that correlates with long-term survival. Therefore it is important clinically to distinguish hibernating from infarcted myocardium, since asynergies due to hibernation will improve on reperfusion, whilst those due to infarct will not. One suggested technique to identify hibernating myocardium is to stimulate the myocytes acutely, but briefly, by administration of inotropic agents while monitoring contractile function by echocardiography. We report our experience on the use of low dosages of dobutamine. Myocardial viability was validated by measuring the recovery in contraction of the akinetic areas after coroanry artery bypass surgery by means of intraoperative epicardial echocardiography. The test has a sensitivity of 93% and a specificity of 78%. It is useful for identification of viable myocardium and also for quantification of intraoperative risk in individual patients. Limitations of this test are related to the presence of downregulation of beta receptors and to the impossibility of differentiating hibernating from stunned myocardium. Another useful technique of identifying hibernating myocardium is the use of radionuclear markers for viability. In our experience the two most important tests are (1) rest-redistribution imaging of thallium 201 (which has a high sensitivity of 93% but a low specificity of 44%) and (2) ^99mTe-Sestamibi imaging, which provides information on both perfusion and function with a single injection. This latter technique allows differentiation between stunning and hibernating on the basis of coronary flow, which is preserved in stunning and reduced in hibernation.     

p38丝裂原活化蛋白激酶信号途径在鼠破骨细胞生成和骨吸收中的作用

目的研究p38丝裂原活化蛋白激酶(p38MAPK)信号途径在甲状旁腺素相关肽(PTHrP)诱导的破骨细胞生成和骨吸收中的作用。方法取小鼠骨髓细胞,在PTHrP(45ng/ml)的刺激下,在不同试验组中分别入0.1、1.0及10μmol/L的p38MAPK抑制剂Fr167653,继续培养6d。抗酒石酸染色,进行破骨细胞计数。在小鼠颅骨部位注射PTHrP建立骨吸收和高钙血症动物模型。每日给予p38MAPK抑制剂Fr16765330mg/kg,每日2次,X线片观察骨吸收面积,组织学检查计算单位面积内破骨细胞数目,采集血样观察全血内游离钙水平。结果PTHrP刺激下,大量破骨细胞生成(118.9±28.3)个/孔;加入0.1μmol/LFr167653可以部分抑制破骨细胞的生成(79.6±28.0)个/孔,加入10μmol/LFr167653几乎全抑制了破骨细胞生成(7.4±0.4)个/孔,每日给予Fr16765330mg/kg,每日2次,可以明显抑制骨吸收,表现为X线片上骨吸收面积减少,单位面积内破骨细胞数目减少,但是并不能有效地抑制高钙血症。结论抑制p38MAPK信号途径可以抑制破骨细胞的分化和局部骨吸收。