内皮损伤相关因子在小鼠肝小静脉闭塞病模型中的表达

随着中草药的广泛应用,近年我国因误服含吡咯烷类生物碱（PAs）的中草药致肝小静脉闭塞病（HVOD）的报道逐渐增多.目的：探讨内皮细胞损伤及其相关因子在HVOD发病中的作用.方法：分别以土三七灌胃30 d和野百合碱灌胃7 d诱导小鼠HVOD模型.动物处死后行血清肝功能指标榆测和全血细胞计数;肝组织切片HE染色、Masson三色染色,行组织学评分;RT-PCR检测肝组织内皮损伤相关因子内皮素-1（ET-1）、肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）以及纤溶酶原激活物抑制剂-1（PAI-1）的表达.结果：土三七组和野百合碱组成模率分别为80.0%和92.0%.两模型组小鼠体质量、肝指数、血清肝功能指标和全血细胞计数的变化以及肝组织学改变符合人类HVOD的临床和病理特征,肝组织ET-1、TNF-α、IL-1β和PAI-1 mRNA表达较正常对照组显著增高（P〈0.05）,其中土三七组PAI-1 mRNA表达显著高于野百合碱组（P〈0.05）.结论：土三七和野百合碱均可成功诱导小鼠HVOD模型,模型小鼠肝组织内皮损伤相关因子表达显著增高,提示内皮细胞损伤后内皮损伤相关因子的释放参与了HVOD的发生过程.     

土三七水煎剂所致肝小静脉闭塞病大鼠模型的建立

目的通过观察不同浓度土三七水煎剂所致的大鼠肝脏病理学变化，建立一种模拟临床患者服药所致肝小静脉闭塞病（HVOD）的动物模型。方法土三七2100g水煎制成浓度为1．5g/ml土三七生药煎剂，SD大鼠76只随机分为A、B、C、D四组，A、B、C组每组20只，分别以土三七水煎剂高（15g·kg^-1·d^-1）、中（7．5g·kg^-1·d^-1）、低（3．75g·kg^-1·d^-1）浓度灌胃，D组16只以温开水灌胃作为对照，各组大鼠雌、雄各半。每日测量大鼠体重并记录异常表现，灌胃1周和2周后各组大鼠分别处死各半，肝脏标本行病理检查，HE及Masson染色，光镜下参照Deleve评分标准判定病变及严重程度。结果土三七高、中、低浓度组分别有83．3％（15／18）、75．0％（15／20）、40．0％（5／20）出现了肝小静脉闭塞病病理表现；相同剂量下雌性较雄性更易发病，雌、雄鼠造模成功率分别为79．3％（23／29）和41．3％（12／29）（P=0．003）。结论土三七水煎剂灌胃可以成功建立HVOD大鼠模型，剂量和大鼠性别与造模成功率直接相关。     

Gynura root induces hepatic veno-occlusive disease： A case report and review of the literature

Gynura root has been used extensively in Chinese folk medicine and plays a role in promoting microcirculation and relieving pain.However,its hepatic toxicity should not be neglected.Recently,we admitted a 62-year old female who developed hepatic veno-occlusive disease(HVOD)after ingestion of Gynura root.Only a few articles on HVOD induced by Gynura root have been reported in the literature.It is suspected that pyrrolizidine alkaloids in Gynura root might be responsible for HVOD.In this paper,we report a case of HVOD and review the literature.     

川芎嗪对脑缺血——再灌注大鼠神经功能及神经电生理的影响

目的观察腹腔注射川芎嗪对脑缺血—再灌注大鼠神经功能及神经电生理的影响。方法将20只大鼠随机分为A、B组,制作大鼠大脑中动脉栓塞模型后,分别腹腔注射生理盐水及川芎嗪80mg／kg,2次／d,连用7d。采用双盲法行大鼠神经功能评分,采用TTC染色法测量大鼠脑梗死灶体积,并行运动诱发电位检测。结果A组神经功能评分为（3．31±0．23）分,脑梗死灶体积为（45．63±12．75）mm^3,运动阈值为52．67％±8．75％,B组分别为（2．30±0．15）分、（26．59±7．64）mm^3、34．29％±5．18％,P均〈0．05。损伤后的运动阈值与脑梗死灶体积呈正相关（r=0．68,P〈0．05）。结论川芎嗪可减小缺血—再灌注损伤脑组织梗死灶体积,提高皮质兴奋性。     

Melatonin ameliorates vascular endothelial dysfunction,inflammation, and atherosclerosis by suppressing the TLR4/NF‐κB system in high‐fat‐fed rabbits

Vascular endothelial dysfunction (VED) and inflammation contribute to the initiation and progression of atherosclerosis. Melatonin (MLT) normalizes lipid profile, improves endothelial function, and possesses anti‐inflammatory properties. However, the precise mechanisms are still unclear. This study investigated whether MLT could ameliorate VED, inflammation, and atherosclerosis by suppressing the Toll‐like receptor 4 (TLR4)/nuclear factor kappa B (NF‐κB) system in high‐fat‐fed rabbits. Rabbits were randomly divided into three groups that received a standard diet (control group), high‐cholesterol diet (atherosclerosis group), or high‐cholesterol diet plus 10 mg/kg/day MLT (MLT group) for 12 wk. After treatment, high‐fat diet significantly increased serum lipid and inflammatory markers in rabbits in atherosclerosis group compared with that in control group. In addition, high‐fat diet also induced VED and typical atherosclerotic plaque formation and increased intima/media thickness ratio, which were significantly improved by MLT therapy as demonstrated in MLT group. Histological and immunoblot analysis further showed that high‐fat diet enhanced the expressions of TLR4, myeloid differentiation primary response protein (MyD88), and NF‐κB p65, but decreased inhibitor of NF‐κB (IκB) expression. By contrast, MLT therapy decreased the expressions of TLR4, MyD88, and NF‐κB p65 and increased IκB expression. This study has demonstrated that MLT ameliorates lipid metabolism, VED, and inflammation and inhibits the progression of atherosclerosis in high‐fat‐fed rabbits. Moreover, our study indicates for the first time that suppression of the TLR4/NF‐κB system in local vasculature with atherosclerotic damage is important for the protective effects of MLT.     

土三七所致肝小静脉闭塞病102例临床分析

目的总结分析土三七所致肝小静脉闭塞病（HVOD）的临床特点,提高对本病的认识。方法对1980年1月至2009年12月国内报道的土三七所致HVOD病例102例进行综合分析,并对国内外相关文献进行复习。结果患者土三七使用量150～5000 g（中位数500 g）,服药时间4 d～1年（中位时间30 d）,临床主要表现为腹胀（99.0%）、腹痛（69.6%）、肝大（93.5%）、腹水（97.7%）、黄疸（63.5%）,有不同程度的肝功能受损。B超检查见肝静脉显示不清,肝段下腔静脉外形变细,但无明显栓塞物,彩色多普勒提示门脉血流减慢或逆向,增强CT示肝脏呈＂地图状＂强化不均匀。病理学表现为肝窦淤血、扩张,流出道受阻,肝细胞不同程度变性、坏死,小叶中央静脉内皮水肿或纤维增生。经内科治疗好转69.7%（62/89）,未愈或死亡30.3%（27/89）,另13例转归不详。使用抗凝或活血化瘀者有效率为81.0%,较未用者（47.8%）,疗效差异有统计学意义（P=0.002）。结论土三七是导致HVOD的一个重要原因,影像和病理检查具有特征性,早期诊断及使用抗凝、活血治疗对改善预后有一定作用。     

JSH-23 targets nuclear factor-kappa B and reverses various deficits in experimental diabetic neuropathy: effect on neuroinflammation and antioxidant defence

Aim: Nuclear factor‐kappa B (NF‐κB) being reported to play an important role in the pathogenesis of diabetic neuropathy is believed to be a central mechanism involved in the genesis and promulgation of inflammatory insult. Here we have targeted the nuclear translocation of NF‐κB using JSH‐23 to elucidate its role in diabetic neuropathy. Methods: JSH‐23 (1 and 3 mg/kg) was administered for 2 weeks in diabetic rats, after 6 weeks of diabetes induction using streptozotocin (55 mg/kg) as diabetogenic agent. Functional (motor nerve conduction velocity and blood flow), behavioural (mechanical hyperalgesia), biochemical [malondialdehyde, glutathione, tumour necrosis factor‐α (TNF‐α) and interleukin‐6 (IL‐6) levels] and NF‐κB translocation studies (western blot technique) were then undertaken. Results: JSH‐23 treatment significantly reversed the nerve conduction and nerve blood flow deficits seen in diabetic animals. Reduction in mechanical pain threshold was also partially corrected by the treatment. Protein expression studies showed that nuclear translocation of p65/p50 subunit was inhibited by JSH‐23 treatment in the sciatic nerve. The treatment also lowered the elevated IL‐6, TNF‐α, cyclo‐oxygenase (COX‐2) and inducible nitric oxide synthase (iNOS) levels/expression, indicating reduction in the inflammatory damage of the sciatic nerve. Apart from these effects, JSH‐23 also increased Nrf2 and hemeoxygenase‐1 (HO‐1) levels which could imply its potential in increasing the strength of antioxidant defence. Conclusion: We observed that NF‐κB inhibition partially reversed functional, behavioural and biochemical deficits with JSH‐23 treatment. This study substantiates the role of NF‐κB activation in the aetiology of diabetic neuropathy and protection afforded by inhibition of NF‐κB by JSH‐23, which can be attributed to its effect on neuroinflammation and oxidative stress.     

Macrolides attenuate mucus hypersecretion in rat airways through inactivation of NF-kappaB

Background and objective: To examine the effect of a 14‐membered ring macrolide on airway mucus hypersecretion in rats treated with LPS. Methods: Mucus hypersecretion in rat airways was induced by intratracheal instillation of LPS. Rats treated with or without LPS were administered roxithromycin (1–10 mg/kg), josamycin (10 mg/kg) or amoxicillin (40 mg/kg), orally for 4 days. Expression of Muc5ac, nuclear factor (NF)‐κB, and the mitogen‐activated protein (MAP) kinases p38 and ERK1/2 in bronchial epithelium were detected by RT‐PCR, immunohistochemistry or western blotting. Mucins, IL‐1β, IL‐8 and tumour necrosis factor (TNF)‐α in BAL fluid were assayed by enzyme‐linked lectin assay and ELISA. Results: LPS significantly induced the expression of Muc5ac mRNA and protein in bronchial epithelium, increased the release of mucins, IL‐1β, IL‐8 and TNF‐α, and increased neutrophil numbers in BAL. Moreover, LPS increased staining for NF ‐ κB in the cytoplasm as well as nuclear translocation of NF ‐ κB in airway epithelial cells. Upregulated expression of Muc5ac mRNA correlated positively with NF ‐ κB activation and the levels of cytokines (P < 0.05). Roxithromycin (5 and 10 mg/kg) significantly attenuated bronchial Muc5ac expression and NF ‐ κB nuclear translocation stimulated by LPS, and reduced neutrophil numbers, mucins and inflammatory cytokines in BAL (P < 0.05). However, LPS‐stimulated expression of p38 and ERK1/2 in airway epithelium was not affected by roxithromycin. Josamycin and amoxicillin had no effects on Muc5ac expression, NF ‐ κB activation or cytokine release. Conclusions: Roxithromycin inhibits the pulmonary inflammatory response and airway mucus hypersecretion induced by LPS. The inhibitory effect of roxithromycin on airway mucus hypersecretion may be mediated through reduction of NF ‐ κB activation, neutrophil infiltration and release of inflammatory cytokines in the lung.     

Effects of gut barrier dysfunction and NF‐κB activation on aggravating mechanism of severe acute pancreatitis

OBJECTIVE: To study the effects of gut‐derived endotoxin translocation and NF‐κB activation on the aggravating mechanism of severe acute pancreatitis (SAP) and of treatment with pyrrolidine dithiocarbamate (PDTC) on rats with SAP. METHODS: SD rats were randomly divided into sham operation group (SO), SAP group, SAP + lipopolysaccharide(LPS) group, pyrrolidine dithiocarbamate (PDTC) treatment group and LPS group. Biochemical parameters and cytokines were examined in the serum. Multiple organs pathological slices were examined. Expression of NF‐κB mRNA in the liver tissue was detected by RT‐PCR. Activation of NF‐κB by the method of streptomycin avidin‐peroxidase (SP) and expression of NF‐κB p65 protein and its binding activity were analyzed by Western blot and electrophoretic mobidity shift assay (EMSA). RESULTS: Compared with sham operation group, the concentration of TNF‐α, alanine aminotransferase (ALT), and diamine oxidase (DAO) in serum significantly increased in SAP + LPS group (P < 0.05). Pathological changes were markedly observed in tissues and the expression of NF‐κB mRNA in the liver significantly increased (P < 0.05) also, the activation of NF‐κB and binding activity of NF‐κB p65 protein in the liver markedly increased (P < 0.01) in SAP + LPS group. Treatment with PDTC markedly reduced concentration of ALT, DAO and TNF‐α, and the expression of NF‐κB, and the pathologic scores, as well as significantly decreased the expression of NF‐κB p65 protein. CONCLUSION: The activation and overexpression of NF‐κB may participate in the aggravating mechanism of SAP. Treatment with PDTC has a protective effect on multiple organs damage in SAP.     

川芎嗪对哮喘大鼠转录因子GATA-3表达的影响

目的 观察川芎嗪对哮喘大鼠转录因子GATA-3表达的影响。方法 72只SPF级SD大鼠随机分为正常对照组（A组）、哮喘模型组（B组）、川芎嗪小剂量组（C组,20mg／kg）、川芎嗪中剂量组（D组,40mg／kg）、川芎嗪大剂量组（E组,80mg／kg）和地塞米松组（F组）,每组12只。以卵蛋白腹腔注射并雾化吸入制备大鼠哮喘模型,用动物呼吸机测定气道反应性评价造模效果。采用免疫组化半定量法测定肺组织GATA-3含量。结果 给予乙酰胆碱（Ach）激发后,比较各组大鼠呼气相气道阻力（Re）,显示造模成功;B、C、D、E和F组的GATA-3表达量显著高于A组,其差异均有统计学意义（P〈0．01）;C、D、E和F组的GATA-3表达量和B组比较,差异均有统计学意义（P〈0．01）;川芎嗪剂量的增加与GATA-3的表达呈负相关趋势;两两比较E组和F组差异无统计学意义。结论哮喘大鼠存在GATA-3高表达;川芎嗪减低气道高反应性,抑制GATA-3的表达,纠正Th1／Th2失衡,从而治疗哮喘。     

川芎嗪对大鼠脑梗塞区细胞Ca~(2 )影响的实验研究

本实验用雄性大白鼠24只随机分成三组:A组(n=8)为对照组,在MCAO前30分钟静注蒸馏水;B组(n=8)在MCAO前30分钟静注川芎嗪lmg/kg;C组(n=8)在MCAD后40分钟静注川芎嗪lmg/kg。用钙离子选择性微电极观察脑梗塞区细胞外Ca~(2 )动态变化。结果显示B组脑梗塞区细胞外Ca~(2 )下降明显低于A组(P<0.05);C组给药前后自身对照脑梗塞区细胞外Ca~(2 )回升不显著(P>0.05)。提示缺血前30分钟用药川芎嗪能预防脑缺血区细胞外Ca~(2 )进入细胞。     

Transcatheter aortic valve implantation for high risk patients With severe aortic stenosis using the Edwards Sapien balloon‐expandable bioprosthesis: A single centre study with immediate and medium‐term outcomes

Background : Transcatheter aortic valve implantation (TCAVI) is an emerging alternative therapy to open‐heart surgery in high‐risk patients with symptomatic aortic stenosis. Methods : Between January 2007 and May 2009, 46 patients underwent TCAVI with the 23 mm or 26 mm Edwards Sapien bioprosthesis via either the transapical (TA‐AVI) or transfemoral (TF‐AVI) approach. All patients had an estimated operative mortality risk of >15%. Results : A total of 46 patients (30 TA‐AVI, 16 TF‐AVI) with a mean aortic valve area (AVA) of 0.63 ± 0.2 cm² and mean gradient of 54 ± 16 mm Hg were treated. Predicted operative mortality was 25.3% by logistic Euroscore and 8.7% by Society of Thoracic Surgeons risk score. Procedural success was 93% in the TA‐AVI group and 88% in the TF‐AVI group. There was one intraprocedural death in the TA‐AVI group. Overall 30‐day mortality was 6.5% (2‐TA‐AVI, 1‐TF‐AVI). Four patients (9.5%) died from noncardiac causes after 30 days. Successful TCAVI was associated with a significant increase in AVA from 0.6 ± 0.1 cm² to 1.6 ± 0.6 cm² in the TA‐AVI group and 0.6 ± 0.1 cm² to 1.4 ± 0.2 cm² in the TF‐AVI group at a mean follow up of 7.4 ± 4.4 and 8.3 ± 5.0 months, respectively. At discharge, there was significant improvement in AVA (P < 0.0001), transaortic mean gradient (P < 0.0001), and mitral regurgitation (P = 0.01). At medium term follow up, the valve area was maintained and there was significant improvement in NYHA class in both groups (P < 0.0001). Conclusion : At medium term follow‐up, both transcatheter approaches demonstrated good valve durability with no cardiac‐related mortality post hospital discharge. © 2009 Wiley‐Liss, Inc.     

Protective effect of exogenous transferrin against hyperoxia: A study on premature rabbits

We hypothesized that an increase in plasma iron binding capacity would decrease the generation of oxygen radicals and of lipid peroxides. To test this hypothesis, we studied whether supplementation of transferrin (TF) in premature rabbits would modify the degree of hyperoxic lung injury. Animals, delivered prematurely at 29 days of gestation (term 31 days), were randomized and given either 0.5 g/kg of albumin (Alb) (n = 116) or 0.5 g/kg of iron-free TF (n = 132) intravenously within 2 hours after birth. Another group was randomized to receive saline (n = 15), or either 0.35 g/kg (n = 12) or 0.70 g/kg of iron-free TF (n = 8). After exposure to a 100% oxygen environment for 2 or 4 days, the animals were killed, and plasma and bronchoalveolar lavage (BAL) fluid was recovered. Infusion of TF caused a dose-dependent increase in the concentration of TF and an increase in the unsaturated iron-binding capacity. Administration of TF at birth increased the gradient of TF between serum and alveolar epithelial lining fluid on day 4, suggesting decreased alveolar-capillary permeability. BAL fluid and plasma from TF-supplemented animals contained less lipid peroxidation products and more inhibitor of lipid peroxidation than BAL fluid or plasma from Alb-treated animals. In TF-treated animals, the recovery of protein in BAL fluid (TF group, 1.26 ± 0.07 mg; Alb group, 1.78 ± 0.10 mg; P = 0.02) and the water content of the extravascular lung tissue (TF group, 78.5 ± 1.4%; Alb group, 83.2 ± 1.3%; P = 0.05) were lower than in Alb-treated animals. We propose that supplementation of iron-free TF decreases iron-catalyzed redox reactions and may decrease hyperoxic lung injury in the premature. Pediatr. Pulmonol. 1997; 24:429–437. © 1997 Wiley-Liss, Inc.     

Combined effects of irbesartan and carvedilol on expression of tissue factor and tissue factor pathway inhibitor in rats after myocardial infarction

The objective of this study was to investigate the effects of irbesartan, carvedilol, and irbesartan plus carvedilol on the expression of tissue factor (TF) and tissue factor pathway inhibitor (TFPI) mRNA and protein in rat myocardium after myocardial infarction (MI). MI was induced in male Wistar rats by ligation of the anterior descending branch of the left coronary artery. Irbesartan at 50 mg/kg/day, carvedilol at 1 mg/kg/day, irbesartan plus carvedilol, or placebo was administered intragastrically; expression of TF and TFPI mRNA and protein was determined by RT-PCR and Western blot analysis. The relative left ventricle weights were lower in all three treatment groups than in the placebo group, with the lowest relative weight in the irbesartan plus carvedilol group (P < 0.001). The size of the infarcted area was lower in the carvedilol and the combined groups than in the placebo group (P < 0.001). The levels of expression of TF and TFPI mRNA and protein were lower in the combined group than in the placebo group or the carvedilol group (P < 0.001). Treatment with irbesartan plus carvedilol reduced the expression of TF and TFPI mRNA and protein after MI in rats, and combined treatment with both agents had greater effects than the single agents alone. These findings suggest that the beneficial effects of these drugs may include anticoagulation and that combined therapy with both agents is an option that should be evaluated further.     

The predictive value and evolution of N-terminal pro-B-type natriuretic peptide levels following transcutaneous aortic valve implantation

Aims: We sought to define the predictive value and evolution of N‐terminal pro‐B‐type natriuretic peptide (NT‐proBNP) levels following transcutaneous aortic valve implantation (TAVI). Methods and Results: We investigated 91 consecutive patients who underwent TAVI (59 transfemoral [TF], 32 transapical [TA]) in our institution. The balloon‐expandable valve was implanted in 75 and the self‐expanding in 16 patients. The baseline (within 48 hours prior to procedure), early (24–74 hours), and late (3–12 months) postprocedural NT‐proBNP levels were determined. The mortality status of all patients was ascertained as of September 2010. The 30‐day and 1.3(mean)‐year mortality was 3% and 12% (2%, 9% in the TF and 6%, 19% in the TA group). Increased baseline (χ²= 5.9, P = 0.016) and early (χ²= 4.9, P = 0.028) NT‐proBNP levels were predictive of mortality. All decrements of the NT‐proBNP levels in the TF patients were significant (baseline 4,984 ± 8,106 vs. early 3,912 ± 6,551 pg/mL, P = 0.016; late 633 ± 606 pg/mL, P = 0.003). In contrast, there was a trend for the early levels to increase in the TA patients (6,423 ± 8,897 vs. 8,100 ± 10,178 pg/mL, P = 0.090), and a significant decline in the late levels as compared to baseline (1,704 ± 3,417 pg/mL, P = 0.005). Conclusion: NT‐proBNP levels are predictive of mortality following TAVI. There is a differential early evolution of their levels between the TF and TA patients and a significant decline later in both groups. (J Interven Cardiol 2011;24:462–469)     

Acute and late outcomes of Transcatheter Aortic Valve Implantation (TAVI) for the treatment of severe symptomatic aortic stenosis in patients at high- and low-surgical risk

Background: This prospective study examines the impact of EuroSCORE and transfemoral (TF) or transapical (TA) delivery approach on mortality at 30 days and 1 year in patients with severe aortic stenosis implanted with either the Edwards SAPIEN Transcatheter Heart Valve (THV) or Medtronic CoreValve. Methods and Results: TAVI was successfully performed in 293 (97.7%) of 300 patients (TF: 174, TA: 126, mean EuroSCORE 24.0). The mortality at 30 days and after 1 year was 6.0% and 17.3%. Mortality depends significantly on the logistic EuroSCORE with a 30‐day odds ratio (OR) of 1.92 (95% CI 1.41 to 2.62, P < 0.001) and after 1 year of 1.67 (95% CI 1.34 to 2.08, P < 0.001). Mortality in patients with a logistic EuroSCORE <15 (n = 113) or ≥15 (n = 187) at 30 days was 0.9% versus 9.1% and after 1 year 7.1% versus 23.5% demonstrating significantly less mortality (P < 0.001) in patients with lower logistic EuroSCOREs. In this specific setup of our center there was no significant difference (P = 0.553) in mortality regarding the technical approach for TA (4.0% and 15.9%) and for TF (7.5% and 18.4%). Severe cardiac complications occurred in 20 patients (6.7%) with a 30‐day mortality of 45.0%. Conclusion: The mortality in patients undergoing TAVI correlates significantly with the logistic EuroSCORE. Patients with a logistic EuroSCORE <15 can be implanted, with a low 30‐day mortality and good long‐term outcome over 1 year. (J Interven Cardiol 2012;25:364–374)     

Antiphospholipid antibodies from patients with the antiphospholipid syndrome induce monocyte tissue factor expression through the simultaneous activation of NF‐κB/Rel proteins via the p38 mitogen‐activated protein kinase pathway,and of the MEK‐1/ERK pathway

Objective

Antiphospholipid syndrome (APS) is characterized by thrombosis and the presence of antiphospholipid antibodies (aPL). In patients with primary APS, expression of tissue factor (TF) on the surface of monocytes is increased, which may contribute to thrombosis in these patients. However, the intracellular mechanisms involved in aPL‐mediated up‐regulation of TF on monocytic cells are not understood. This study was undertaken to investigate the intracellular signals induced by aPL that mediate TF activation in monocytes from APS patients.

Methods

We analyzed, both in vivo and in vitro, aPL interactions with proteins that have signaling functions, including mitogen‐activated protein kinases (MAP kinases) and NF‐κB/Rel proteins.

Results

In vivo studies demonstrated significantly higher levels of both TF messenger RNA and TF protein in monocytes from APS patients compared with controls. At the molecular level, increased proteolysis of IκBα and activation of NF‐κB were observed. Constitutive activation of both p38 and ERK‐1 MAP kinases was also found. Treatment of normal monocytes with aPL activated ERK‐1 and p38 MAP kinases, as well as the IκB/NF‐κB pathway, in a dose‐dependent manner. NF‐κB activation and IκBα degradation induced by aPL were inhibited by the NF‐κB inhibitor SN50 and the p38 MAP kinase inhibitor SB203580, thus suggesting crosstalk between these pathways. However, the MEK‐1/ERK inhibitor PD98059 did not affect aPL‐induced NF‐κB binding activity. TF expression induced by aPL was significantly inhibited by combined treatment with the 3 inhibitors.

Conclusion

Our results suggest that aPL induces TF expression in monocytes from APS patients by activating, simultaneously and independently, the phosphorylation of MEK‐1/ERK proteins, and the p38 MAP kinase–dependent nuclear translocation and activation of NF‐κB/Rel proteins.

     

Combined melatonin and exendin‐4 therapy preserves renal ultrastructural integrity after ischemia–reperfusion injury in the male rat

We tested whether combined melatonin (Mel) and exendin‐4 (Ex4) treatment can better preserve glomerular structural integrity after ischemia–reperfusion (IR) injury compared with either alone. Adult male Sprague Dawley rats (n = 50) were equally divided into sham control (SC), IR, IR‐Ex4 (10 μg/kg subcutaneously 30 min after reperfusion and daily for 5 days), IR‐Mel (20 mg/kg intraperitoneally at 30 min postreperfusion and 50 mg/kg at 6 and 18 hr), and IR‐Ex4‐Mel were euthanized at day 14. Serum creatinine level and urine protein‐to‐creatinine ratio at days 3 and 14 were highest in IR group and lowest in SC, significantly higher in IR‐Ex4 and IR‐Mel groups than in IR‐Ex4‐Mel group (all P < 0.001) without significant difference between IR‐Ex4 and IR‐Mel groups. Changes in podocyte injury score (PIS) and kidney injury score were highest in IR group and lowest in SC, significantly higher in IR‐Ex4 and IR‐Mel groups than in IR‐Ex4‐Mel, and significantly higher in IR‐Mel group than in IR‐Ex4 group (all P < 0.001). Immunohistochemical microscopic findings of the expressions of FSP‐1 and WT‐1 (two glomerular damage indicators) and KIM‐1 and snail (two renal tubular‐damaged indicators) showed an identical pattern, whereas the expressions of ZO‐1, p‐cadherin, podocin, dystroglycan, fibronectin, and synaptopodin (six indices of glomerular integrity) demonstrated an opposite pattern compared to that of PIS among five groups (all P < 0.001). Protein expressions of inflammatory (TNF‐α/NF‐κB/MMP‐9) and oxidative stress (NOX‐1, NOX‐2, oxidized protein) biomarkers exhibited an identical pattern to that of PIS among five groups (all P < 0.001). Combined melatonin–exednin‐4 therapy further protected glomerulus from IR injury.