Histamine differentially interacts with tumor necrosis factor-α and thrombin in endothelial tissue factor induction: the role of c-Jun NH2-terminal kinase

BACKGROUND: Histamine plays an important role in vascular disease. Tissue factor (TF) expression is induced in vascular inflammation and acute coronary syndromes. OBJECTIVES: This study examined the effect of histamine on tumor necrosis factor-alpha- (TNF-alpha-) vs. thrombin-induced endothelial TF expression. METHODS AND RESULTS: Histamine (10(-8)-10(-5) mol L-1), TNF-alpha (5 ng mL-1), and thrombin (1 U mL-1) induced TF expression in human endothelial cells. Although TF expression by TNF-alpha and thrombin was identical, histamine augmented TNF-alpha-induced expression 7.0-fold, but thrombin-induced expression only 2.6-fold. Similar responses occurred with TF activity. The H1-receptor antagonist mepyramine abrogated these effects. Differential augmentation by histamine was also observed at the mRNA level. Histamine-induced p38 activation preceded a weak second activation to both TNF-alpha and thrombin. Histamine-induced c-Jun NH2-terminal kinase (JNK) activation was followed by a strong second activation to TNF-alpha, and less to thrombin. Selective inhibition of this second JNK activation by SP600125 reduced TF induction to histamine plus TNF-alpha by 67%, but to histamine plus thrombin by only 32%. Histamine augmented TNF-alpha- and thrombin-induced vascular cell adhesion molecule 1 (VCAM-1) expression to a similar extent. Consistent with this observation, VCAM-1 induction to TNF-alpha and thrombin was mediated by p38, but not by JNK. CONCLUSIONS: Histamine differentially augments TNF-alpha- vs. thrombin-induced TF expression and activity, which is mediated by the H1-receptor, occurs at the mRNA level, and is related to differential JNK activation.     

Total and free tissue factor pathway inhibitor in pregnancy hypertension.

OBJECTIVE: To clarify the role played by tissue factor pathway inhibitor (TFPI) in pregnancy hypertension. METHODS: Using enzyme-linked immunosorbent assays, hemostatic measurements were obtained for women with pre-eclampsia (n=51), nonproteinuric hypertension of pregnancy (n=62), postpartum pre-eclampsia 24 h after childbirth (n=31), and no hypertension (healthy pregnant controls, n=100). RESULTS: There was a significant increase in circulating free TFPI levels in women with pre-eclampsia (9.7+/-6.2 ng/mL) or nonproteinuric hypertension of pregnancy (8.3+/-5.3 ng/mL) compared with healthy controls (5.3+/-2.1 ng/mL). In women with pre-eclampsia the levels remained elevated after placental delivery (10.6+/-4.0 ng/mL). Free protein S levels were significantly higher in women with pre-eclampsia (40.0%+/-10.7%), nonproteinuric hypertension of pregnancy (37.1%+/-12.5%), or postpartum pre-eclampsia (39.3%+/-9.1%) than in healthy pregnant controls (32.2%+/-8.5%). CONCLUSION: Increased levels of the physiologically active free forms of TFPI and free protein S, 2 coagulation inhibitors, may protect women with pregnancy-induced hypertension from the risks of hemostatic activation.     

PGE1 abolishes the mitochondrial-independent cell death pathway induced by D-galactosamine in primary culture of rat hepatocytes

Background and Aim:  PGE₁ reduces in vivo and in vitro D-galactosamine (D-GalN)-induced cell death in hepatocytes. The present study was undertaken to elucidate the intracellular pathway by which D-GalN induces cell death in cultured hepatocytes. In addition, we evaluated if PGE₁ was able to modulate different parameters related to D-GalN-induced apoptosis in cultured rat hepatocytes.
Methods:  Hepatocytes were isolated from male Wistar rats (225–275 g) by the classical collagenase procedure. PGE₁ (1 µM) was administered 2 h before D-GalN (5 mM) in primary culture of rat hepatocytes. Apoptosis was determined by DNA fragmentation and caspase-3, -6, -8 and -9 activation in hepatocytes. Caspase activation was evaluated by the detection of the related cleaved product and its associated activity. Cell necrosis was determined by the measurement of lactate dehydrogenase (LDH) activity in culture medium. To elucidate the role of mitochondria, we measured neutral (nSMase) and acid (aSMase) sphingomyelinase, as well as the expression of cytochrome c in mitochondria and cytoplasm fractions from D-GalN treated hepatocytes.
Results:  D-GalN induced caspase-3 activation and DNA fragmentation in hepatocytes. This apoptotic response was not associated with the activation of caspase-6, -8 or -9. The use of specific inhibitors confirmed that only caspase-3 was involved in D-GalN-induced apoptosis. D-GalN did not modify nSMase and aSMase activities, nor mitochondrial cytochrome c release in hepatocytes.
Conclusions:  D-GalN induced apoptosis through caspase-3 activation but without modification of the activity of caspase-6, -8, -9, SMases or cytochrome c release. PGE₁ appears to prevent D-GalN-induced apoptosis by a mitochondria-independent mechanism.     

不同浓度的富马毒素B1对肝癌细胞凋亡的影响

目的探讨不同浓度富马毒素对肝癌细胞凋亡的影响,为进一步研究富马毒素诱导凋亡的机制打下基础。方法利用PI标记的流式细胞术测定不同浓度下富马毒素对体外培养肝癌细胞凋亡率的影响。结果在1.25nmol/ml的富马毒素作用下凋亡率与空白没有差异,在2.5～40nmol/mlFB1的浓度范围内出现了明显的凋亡,随着富马毒素浓度的增加,体外培养的肝细胞凋亡率增加,且凋亡率在15nmol/ml达到高峰。结论富马毒素可导致肝癌细胞凋亡,随着浓度的递增,其凋亡率也随之递增。     

白细胞介素6、信号传导和转录活化因子3和血管内皮生长因子在人脑胶质瘤中的表达及相关性研究

目的研究人脑不同级别胶质瘤中白细胞介素（IL）-6，信号传导和转录活化因子3（STAT3）和血管内皮生长因子（VEGF）的表达，探讨IL-6、STAT3和VEGF与肿瘤病理级别和侵袭性的关系。方法采用免疫组织化学法，检测70例人脑胶质瘤，10例脑膜瘤和5例正常脑组织中IL-6、STAT3和VEGF的表达。结果胶质瘤中IL-6、STAT3和VEGF的表达水平在高级别组（Ⅲ、Ⅳ级）明显高于低级别组（Ⅰ、Ⅱ级），两组间差异有统计学意义（P〈0．01），STAT3的表达与IL-6和VEGF的表达均呈正相关（P〈0．01）。结论IL-6、STAT3和VEGF的表达与胶质瘤的恶性程度有密切关系；且三者协同在胶质瘤发生、发展过程中起重要作用。三者的相关性证实VEGF基因由STAT3蛋白调节，而STAT3又由IL-6刺激活化。     

Is there a ‘Basque’ profile regarding autosomal recessive deficiencies of coagulation factors?*

When excluding haemophilia and von Willebrand disease, coagulation factors deficiencies constitute rare autosomal recessive disorders (<1 in 500,000) of less precisely defined epidemiology. We have reported herein the distribution of these entities in the French Basque Country, a genetic isolate of very old individualization with peculiar biological specificities. The prevalence of these disorders was markedly high, especially, as already shown, factor XI deficiency. This unusual profile needs to be discussed in the view of population genetics.     

急性胰腺炎患者vW因子的变化及其临床意义

目的　探讨急性胰腺炎患者血浆vW因子的变化及其临床意义。方法　采用双抗体夹心固相酶免疫法测定 3 5例急性胰腺炎患者和 3 0名健康献血者的血浆vW因子水平 ,对其进行相关性分析。结果　急性胰腺炎患者入院后 2 4h血浆vW因子水平为 2 15 .0 %± 2 3 .4% ,显著高于正常对照组 10 7.5 %± 2 9.6% (P <0 .0 0 1) ;重症急性胰腺炎患者血浆vW因子显著高于轻型急性胰腺炎患者 (P <0 .0 1) ;血浆vW因子预测重症AP的敏感性、特异性和准确性分别是 91.7%、87.0 %和 88.7%。结论　急性胰腺炎患者存在血管内皮损伤 ;测定急性胰腺炎患者血浆vW因子水平可早期预测病情的严重程度     

非诺贝特对培养的兔脂肪细胞表达凝血和纤溶因子的影响

目的探讨非诺贝特对培养的兔脂肪细胞表达组织因子(TF)和纤溶酶原激活物抑制剂1(PAI-1)的影响.方法取正常兔脂肪组织分离培养脂肪细胞,以不同浓度(分别为0,1,10和100μmol/L)非诺贝特孵育兔脂肪细胞24 h后收集细胞.RT-PCR测定脂肪细胞TF和PAI-1 mRNA表达.用ELISA方法测定TF和PAI-1浓度.结果不同浓度非诺贝特均可抑制兔脂肪组织细胞TF和PAI-1的表达和蛋白产生,其抑制作用呈浓度依赖性增强.在非诺贝特10μmol/L培养时兔脂肪细胞TF和PAI-1 mRNA分别为(0.504±0.016)和(1.500±0.096),明显低于对照[分别为(0.579±0.018)和(1.607±0.063),均P＜0.01].在非诺贝特100μmol/L培养时兔脂肪细胞TF和PAI-1 mRNA分别为(0.451±0.023)和(1.269±0.084),与对照比显著降低(均P＜0.001).结论非诺贝特能抑制兔脂肪细胞TF和PAI-1 mRNA和蛋白的表达,提示非诺贝特可能具有独立于降脂作用外的抗血栓作用.     

常温与低温体外循环心脏直视手术对细胞因子及补体的影响

目的探讨常温及低温体外循环心脏直视手术对细胞因子及补体的影响。方法选择先天性和风湿性心脏病患者40例,随机分为常温组及低温组各20例,分别于术晨、体外循环结束时及术后1、4、7、14 d抽取患者静脉血标本,测定血浆TNF、IL-2、C3、C4值。结果两组术前各项检查指标无显著差异。(1)两组术后1~4 d的IL-2水平较术前显著下降,至术后7 d恢复正常。体外循环结束至术后4 d,低温组IL-2显著低于常温组。(2)体外循环结束时以及术后1、4、7 d,常温组TNF水平显著低于低温组。两组体外循环结束时及术后1、4 d均高于术前,常温组至术后7 d、低温组术后14 d恢复正常。(3)体外循环结束时及术后1、7 d,常温组C3水平高于低温组,术后4 d两组无差别;常温组及低温组于体外循环结束时、术后1、4 d均低于术前,至术后7 d常温组恢复正常,低温组至术后14 d恢复至术前水平。(4)两组体外循环结束时及术后1、4 d C4水平均低于术前。体外循环结束时、术后1 d,常温组C4水平高于低温组。结论常温体外循环心脏直视手术对细胞因子及补体的影响显著轻于低温组,因而对术后机体的恢复优于低温方法。