Antiendothelial cell antibodies in lupus: correlations with renal injury and circulating markers of endothelial damage

Systemic lupus erythematosus is characterized by the productionof a broad spectrum of autoantibodies. Autoantibodies directedagainst endothelial cells (AECA) have been particularly welldocumented. We investigated associations between such antibodies,double-stranded DNA (dsDNAb), phospholipid (cardiolipin, ACA),and indices of activity and chronicity scored on renal biopsyspecimens from 22 patients with acute lupus. AECA were presentin 73% of these patients, and both the percentage of patientswith AECA and the mean antibody titre fell significantly aspatients entered remission. When patients already on immunosuppressivetherapy were excluded from analysis (n = 7), only levels ofAECA and DNAb (p = 0.02) correlated with histological evidenceof active lesions and the presence of glomerular epithelialcell crescents; no correlation was found with chronic changesin the renal biopsies. Serum von Willebrand factor (vWf) andserum total protein S levels, two parameters reflecting endothelialcell function, were also measured during acute disease and remission.vWf concentrations were elevated during acute disease (m = 1.9lU/ml, p = 0.02), but the values did not correlate with AECAtitres. In contrast, total protein S levels were reduced (0.81lU/ml vs. 0.97 lU/ml, p = 0.01) during active disease, but remainedwithin the normal range (0.56–1.16 lU/ml). Furthermore,protein S levels were inversely related to levels of AECA (r= –0.4, p = 0.01). AECA were therefore present in mostpatients with acute lupus nephritis and were associated withhistological evidence of active renal injury and serologicalevidence of endothelial cell dysfunction. These data provideindirect support for a pathogenic role for AECA in lupus nephritis.     

探讨几种血清学指标联合检测在系统性红斑狼疮早期肾损伤中的价值

目的探讨血清免疫球蛋白(IgG、IgA、IgM)、补体(C3、C4)及胱抑素C(Cys-C)联合检测在系统性红斑狼疮(SLE)早期肾损害的临床应用价值。方法在AU2700全自动生化分析仪上对该院2012年3月至2014年2月的72例SLE患者,根据24h尿微量清蛋白排泄率(UAER)分成UAER正常组、UAER微量组、UAER过量组与32例健康体检的对照组,同步进行血清IgG、IgA、IgM、C3、C4、Cys-C、尿素(Urea)和肌酐(Cr)指标检测,并对检测结果进行统计学分析。结果 UAER正常组血清IgG、IgA、IgM、Cys-C水平均高于对照组,C3、C4水平低于对照组;UAER微量组各指标水平高于对照组。UAER正常组和UAER微量组血清IgG、IgA、IgM、C3、C4和Cys-C阳性检出率分别为(37.5%、70.4%)、(29.2%、66.7%)、(33.3%、55.6%)、(45.8%、74.1%)、(41.7%、63.0%)、(50.0%、77.8%),高于Urea(8.3%,29.6%)和Cr(12.5%,25.9%),组间比较差异有统计学意义(P0.05)。UAER过量组血清IgA、IgM、Urea和Cr水平高于对照组,组间比较差异有统计学意义(P0.05);UAER过量组血清IgG和Cys-C水平明显高于对照组,C3、C4水平明显低于对照组,组间比较差异有统计学意义(P0.05),UAER过量组各指标的阳性检出率均超过90.0%,各指标比较差异无统计学意义(P0.05)。结论联合检测血清Ig、补体和Cys-C水平可作为SLE早期肾损害诊断依据,对探讨SLE所致肾损害的发病机制、早期诊断、病情进展判断以及治疗预后观察等方面具有重要的临床意义。     

脓毒症时内皮细胞激活、功能障碍和损伤相关标志物的检测

脓毒症作为一种临床综合征,是重症监护病房(ICU)患者死亡的第1大原因,居所有死亡原因的前10位[1]。现已认识到是机体的全身炎症反应综合征(SIRS)而非感染源本身才是影响脓毒症患者结局的关键。循环内皮细胞(CEC)不但构成了血液和组织之间的屏障,而且在炎症、免疫、凝血、抗凝、纤     

Acute myocardial infarction is associated with endothelial glycocalyx and cell damage and a parallel increase in circulating catecholamines

Introduction

Excessive sympathoadrenal activation in critical illness contributes directly to organ damage, and high concentrations of catecholamines damage the vascular endothelium. This study investigated associations between potential drivers of sympathoadrenal activation, circulating catecholamines and biomarkers of endothelial damage and outcome in ST segment elevation myocardial infarction (STEMI)-patients, hypothesizing that the catecholamine surge would reflect shock degree and correlate with biomarkers of endothelial damage.

Methods

This was a prospective study of 678 consecutive STEMI-patients admitted to a single high-volume invasive heart centre for primary percutaneous coronary intervention (pPCI) from September 2006 to July 2008. Blood samples were drawn immediately before pPCI. Plasma adrenaline, noradrenaline, syndecan-1 and thrombomodulin were measured retrospectively with complete data in 571 patients (84%). Median follow-up time was 28 (IQR 23 to 34) months. Follow-up was 99.7% complete. Outcomes were all-cause and cardiovascular mortality, re-myocardial infarction and admission due to heart failure.

Results

Circulating noradrenaline and adrenaline correlated weakly but independently with syndecan-1 (rho = 0.15 and rho = 0.13, both P <0.01) and thrombomodulin (rho = 0.11 and rho = 0.17, both P <0.01), biomarkers of glycocalyx and endothelial cell damage, respectively. Considering biomarkers, patients with shock pre-pPCI had higher adrenaline and syndecan-1 and patients admitted to ICU post-pPCI had higher syndecan-1 (all P <0.05), and in the patients with shock (n = 51) catecholamines correlated strongly with thrombomodulin and syndecan-1 (rho = 0.31 to 0.42, all P <0.05). During follow-up, 78 (14%) patients died (37 cardiovascular deaths) and 65 (11%) were admitted with heart failure. By multivariate Cox proportional hazards analyses, one quartile higher plasma adrenaline was weakly but independently associated with both 30-day and long term mortality and heart failure (30-day all-cause mortality hazard ratio (95% CI) 1.39 (1.01 to 1.92), P = 0.046; 30-day heart failure 1.65 (1.17 to 2.34), P = 0.005; and long-term cardiovascular mortality 1.49 (1.08 to 2.04), P = 0.014). Furthermore, one quartile higher syndecan-1 was also weakly but independently associated with long-term all cause mortality (1.26 (1.02 to 1.57), P = 0.034).

Conclusions

In STEMI patients treated with pPCI, catecholamines correlated weakly with biomarkers of endothelial damage, with the strongest correlations and highest adrenaline and syndecan-1 levels in patients with shock. Furthermore, adrenaline and syndecan-1 were weakly but independently associated with mortality and heart failure. Acute myocardial infarction appears to cause significant endothelial cell and glycocalyx injury and a parallel increase in circulating catecholamines.     

Elevation of circulating endothelial microparticles in patients with chronic renal failure

BACKGROUND: Chronic renal failure patients are at high risk of cardiovascular events and display endothelial dysfunction, a critical element in the pathogenesis of atherosclerosis. Upon activation, the endothelium sheds microparticles, considered as markers of endothelial dysfunction that also behave as vectors of bioactive molecules. AIM: To measure plasma levels of endothelial microparticles (EMPs) in chronic renal failure patients (CRF), either undialyzed or hemodialyzed (HD), and to investigate the ability of uremic toxins to induce EMP release in vitro. METHODS: Circulating EMPs were numerated by flow cytometry, after staining of platelet-free plasma with phycoerythrin (PE)-conjugated anti-CD144 (CD144+ EMP) or anti-CD146 (CD146+ EMP) monoclonal antibodies. Platelet MP (CD41+ PMP), leukocyte MP (CD45+ leukocyte microparticles (LMP)), and annexin-V+ MPs were also counted. In parallel, MPs were counted in supernatant of human umbilical vein endothelial cells incubated with uremic toxins [oxalate, indoxyl sulfate, p-cresol, and homocysteine (Hcy)], at concentrations found in patients. RESULTS AND CONCLUSIONS: CD144+ EMP and CD146+ EMP levels were significantly higher in CRF and HD patients than in healthy subjects. Furthermore, annexin-V+ MPs were elevated in both groups of uremic patients, and CD41+ PMP and CD45+ LMP were increased in CRF and HD patients, respectively. In vitro, p-cresol and indoxyl sulfate significantly increased both CD146+ and annexin-V+ EMP release. Increased levels of circulating EMP in CRF and HD patients represent a new marker of endothelial dysfunction in uremia. The ability of p-cresol and indoxyl sulfate to increase EMP release in vitro suggests that specific uremic factors may be involved in EMP elevation in patients.     

Predisposing factors to thrombosis in systemic lupus erythematosus: possible relation to endothelial cell damage.

We studied 28 patients with SLE, five of whom had had thrombotic episodes. Platelet function and coagulation and fibrinolytic studies were performed to determine whether any of these factors may predispose to thrombosis in SLE. An inhibitor of blood coagulation was detected in 12 patients, and von Willebrand's syndrome was observed in two others. The most striking findings which could be correlated with thromboembolic phenomena were the increase in VII R:WF and the absence of plasminogen activator release after venous occlusion. Both proteins are synthesized by the endothelial cell, and the abnormalities observed are possibly related to damage of the vascular endothelium by immune complexes observed in SLE.     

Pulmonary endothelial permeability and circulating neutrophil-endothelial markers in patients undergoing esophagogastrectomy

OBJECTIVE: Esophagogastrectomy is an established surgical treatment for esophageal malignancy. The postoperative period may be complicated by the development of acute lung injury syndromes and thus, may provide a useful model in which to study the early pathogenic mechanisms of inflammatory lung injury. DESIGN: Open, prospective study. SETTING: High dependency and intensive therapy units. PATIENTS: Eight healthy male volunteers and 20 patients in the early postoperative period INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The lung protein accumulation index (PAI) of radiolabeled transferrin was determined by using a portable, double-isotope system. The following circulating inflammatory markers-thought to reflect neutrophil-endothelial activation and injury including circulating neutrophil elastase-soluble L-, E-, and P-selectins and thrombomodulin and von Willebrand factor antigen were assayed from venous blood samples The PAI for healthy volunteers was median -0.5 (range, -1.73 to 0.27) x 10(-3)/min and for patients undergoing esophagogastrectomy -0.005 (range, -1.53 to 2.28) x 10(-3)/min. There was no statistical difference between the two groups. In the postesophagogastrectomy group, a significant elevation in circulating levels of neutrophil elastase, soluble P- and E-selectin, thrombomodulin, and von Willebrand factor antigen were observed relative to the control group but only circulating plasma elastase demonstrated a significant correlation with the PAI (r2 = .23, p =.03). CONCLUSIONS: The data suggest patients undergoing esophagogastrectomy develop a inflammatory response but this is not a surrogate of permeability and other factors are likely to determine persistent injury to the alveolar-capillary barrier function in this patient group.     

血管内皮细胞损伤性疾病患者抗内皮细胞抗体和抗心磷脂抗体检测及其意义

目的探讨抗内皮细胞抗体(AECA)及抗心磷脂抗体(ACA)与血管内皮细胞损伤性疾病的关系及意义。方法建立ELISA法检测AECA,采用ELISA法测定ACA,分别检测168例血管内皮细胞损伤性疾病患者中2种抗体的水平。结果168例血管内皮细胞损伤性疾病中冠心病、特发性血小板减少性紫癜、溶血性尿毒综合征及妊娠高血压综合征(妊高征)患者的AECA和ACA的阳性率分别为32.9%、22.7%、22.2%、38.9%和38.2%、31.8%、33.3%、44.4%,均显著高于正常对照组。结论AECA和ACA与血管内皮细胞损伤性疾病有一定的相关性。AECA与ACA无相关性。     

Association of anticardiolipin antibodies with intraglomerular thrombi and renal dysfunction in lupus nephritis

We studied positivity for anti-cardiolipin antibody, intraglomerular capillary thrombi on renal biopsy, and the progression of renal disease in 51 patients (10 male and 41 female), mean age 37 years (range 17-65 years), with a diagnosis of systemic lupus erythematosis and clinically evident nephritis confirmed by renal biopsy. Serum creatinine, serum indicators of disease activity and biopsies were analysed in subgroups according to thrombi and anticardiolipin status. End-points were death or chronic dialysis requirement and survival. Degree of sclerosis, crescent formation and necrosed glomeruli were all greater in those specimens positive for thrombi and in those specimens of patients who were serum ACA-positive, suggesting a relationship to disease activity/severity at presentation. The increase in serum anti-DNA antibodies and ANA and the reduction in C3 and C4 were significant in ACA-positive patients, with a strong relationship to disease activity when compared with changes in the ACA-negative patients (p < 0.05 in all cases). There was no significant difference when patients were separated according to the presence or absence of thrombi. Renal function at presentation was worse in patients with intracapillary thrombi and ACA positivity (p = 0.085 and p = 0.042, respectively). All patients progressed, but only those with intracapillary thrombi or anti- cardiolipin antibody positivity had a significant deterioration in renal function. Twenty-one thrombotic episodes occurred in 14 patients, of whom 13 were ACA-positive. Anticardiolipin antibody is a strong predictor of the presence of intraglomerular thrombi in SLE patients with renal involvement. The presence of thrombi and/or anticardiolipin antibodies indicate a worse long-term renal outcome. Anti-cardiolipin antibody positivity is a strong predictor of systemic vascular thrombotic complications.      

系统性红斑狼疮患者凝血纤溶异常与肾损害的相关性研究

目的 探讨系统性红斑狼疮(SLE)患者凝血纤溶异常与肾损害的关系.方法 分别检测比较53例SLE患者与30例同期健康体检者的24小时尿总蛋白、尿微量白蛋白、尿β2微球蛋白及D-二聚体,并进行相关性分析.结果 有肾损害组D-二聚体、尿微量白蛋白、尿β2微球蛋白均高于无肾损害组,其D-二聚体水平与尿微量白蛋白、尿β2微球蛋白呈正相关,治疗后三者均下降.结论 SLE患者大多数D-二聚体水平升高且多数有肾损害.除及时原发病治疗外,适当抗凝治疗,有利于改善病情,减少肾损害.     

Detection of anti-vascular endothelial cell antibodies with microcytotoxicity testing

Antivascular endothelial cell antibodies (anti-VEC) were detected in 20 out of 20 serum samples from post renal transplant nephrectomy patients using a microcytotoxicity (MET) test, but in only 1 of 100 healthy blood donors. Cytotoxicity to VEC could occur in the absence of lymphocytotoxic antibodies. In this paper factors influencing the specificity and sensitivity of microcytotoxicity on vascular endothelial cell (VEC) were studied, including improvements in the preparation of VEC from an umbilical cord vein to get 95% and more of purity and viability; adequate dilution of rabbit sera to reduce its nonspecific VEC cytotoxic effect; and results read by exactly adjusted phase contrast microscopy to reduce the percentage of false negative. The original titers of allotypic and monoclonal antibodies against VEC have been shown to be reproducible in repeated testing during the past two years. The recognition of the weak cytotoxic effects of anti-HLA on VEC makes possible direct application of microcytotoxicity on VEC, to detect anti-VEC and to study VEC antigen classification (through a comparison of the cytotoxic effects of tested sera on VEC and concordant lymphocytes).     

肾综合征出血热患者血清视黄醇结合蛋白和胱抑素C水平及其与肾损伤的相关性分析

目的 探讨肾综合征出血热患者血清视黄醇结合蛋白(RBP)和胱抑素C(CysC)水平及其与肾损伤的相关性.方法 选取肾综合征出血热患者150例作为实验组.选择体检健康者150例作为对照组.对比2组RBP、CysC、尿素及肌酐水平;分析实验组在不同时期RBP、CysC、尿素及肌酐检测阳性率;分析RBP、CysC与尿素及肌酐相关性;分析RBP与CysC单项检测及联合检测的阳性率.结果 与对照组相比,实验组在发热期RBP、CysC、尿素及肌酐水平显著升高(P<0.05).实验组在低血压休克期、少尿期及多尿期4项指标检测阳性率最高.RBP、CysC与尿素及肌酐均呈正相关(r=0.76、0.81、0.89、0.84,P<0.05).联合检测在不同时期的检测阳性率均显著高于单一检测(P<0.05).结论 RBP、CysC均与尿素及肌酐呈正相关,且联合检测具有较高的诊断符合率.     

Semicarbazide-sensitive amine oxidase in pre-eclampsia: no relation with markers of endothelial cell activation

BACKGROUND: Semicarbazide sensitive amine-oxidase (SSAO) is an adhesion molecule and thought to play a role in endothelial cell dysfunction (ECD). SSAO has never been associated with markers of ECD. Pre-eclampsia (PE) is, like the early stages of atherosclerosis, characterised by ECD. SSAO could contribute to ECD in PE. METHODS: Plasma samples were obtained in 14 pre-eclamptic patients and 14 matched controls. In these SSAO-activity, von Willebrand factor (vWF) levels and ED1 fibronectin levels were determined. Placental tissue was collected of 12 pre-eclamptic pregnancies, 8 preterm deliveries and 12 term controls. In these samples, SSAO activity was assessed. In a subset of these placentas, immunohistochemical staining was performed for SSAO, ICAM-1 and VCAM-1. RESULTS: Plasma SSAO activity was not significantly different between pre-eclamptic subjects and controls. VWF and ED1 fibronectin were both significantly increased in the pre-eclamptic subjects. There was no correlation between SSAO activity and vWF or ED1 fibronectin levels. Placental SSAO activity was not different between pre-eclamptic pregnancies, preterm deliveries and term controls. There was strong staining of SSAO in vascular smooth muscle cells (VSM) and moderate staining of trophoblast in all three groups. Endothelial cell expression of SSAO was only seen in term controls and the placentas of pre-eclamptic patients. There was no association between the expression of SSAO, ICAM-1 or VCAM-1 in the placentas of pre-eclamptic patients. CONCLUSION: SSAO expression and activity are not related to markers of ECD.     

Endonuclease-induced DNA damage and cell death in oxidant injury to renal tubular epithelial cells.

Hydrogen peroxide (H2O2)-induced DNA damage and cell death have been attributed to the direct cytotoxicity of H2O2 and other oxidant species generated from H2O2. We examined the possibility that oxidants activate endonucleases leading to DNA damage and cell death in renal tubular epithelial cells, similar to that described for apoptosis. Within minutes, H2O2 caused DNA strand breaks in a dose-dependent manner, followed by cell death. DNA fragmentation was demonstrated both by the release of [3H]thymidine in 27,000-g supernatant as well as the occurrence of low molecular weight DNA fragments on agarose gel electrophoresis, characteristic of endonuclease cleavage. Endonuclease inhibitors, aurintricarboxylic acid, Evans blue, and zinc ion prevented H2O2-induced DNA strand breaks, fragmentation, and cell death. Inhibitors of protein or mRNA synthesis had only minor protection against H2O2-induced DNA damage in contrast to complete protection reported in apoptotic thymocytes. Micrococcal endonuclease induced similar DNA strand breaks in LLC-PK1 cells, and the endonuclease inhibitors prevented the events confirming the ability of endonucleases to induce DNA damage. The protective effect of aurintricarboxylic acid was not due to the prevention of the rise in intracellular free calcium. We conclude that endonuclease activation occurs as an early event leading to DNA damage and cell death in renal tubular epithelial cells exposed to oxidant stress and, in contrast to apoptotic thymocytes, does not require macromolecular synthesis.     

尿mALB/Cr和尿NAG/Cr检测在系统性红斑狼疮患者早期肾损害监测中的临床意义

目的探讨尿N-乙酰β-D氨基葡萄糖苷酶/尿肌酐(UNAG/Cr)、尿微量清蛋白/尿肌酐(UmALB/Cr)在系统性红斑狼疮患者早期肾损害监测中的临床意义。方法 47例系统性红斑狼疮患者尿蛋白定性阴性组,50例系统性红斑狼疮患者尿蛋白定性阳性组及40例健康对照组:采用免疫比浊法测定UmALB;终点法检测尿NAG;速率法检测尿Cr。结果系统性红斑狼疮患者尿蛋白定性阴性组UNAG/Cr、UmALB/Cr高于健康对照组(P<0.01);系统性红斑狼疮患者尿蛋白定性阳性组UNAG/Cr、UmALB/Cr明显高于健康对照组及尿蛋白定性阴性组(均P<0.01)。结论 UNAG/Cr、UmALB/Cr是诊断系统性红斑狼疮患者肾脏早期损伤敏感的指标。     

抗心磷脂抗体与狼疮性肾炎患者肾功能的关系

目的探讨狼疮性肾炎患者抗心磷脂抗体(ACA)对肾功能的影响。方法对68例狼疮性肾炎患者随访1年,测定其血清ACA及肾功能。结果狼疮性肾炎肾功能正常组和肾功能不全组的IgG鄄ACA、IgA鄄ACA、IgM鄄ACA阳性率分别为40.5%、35.7%、40.5%(P<0.01)及57.7%、38.5%、46.2%(IgG,P<0.01;IgA,IgM,P<0.05),与对照组相比差异有显著性。随访期ACA阳性组肾功能不全者明显超过ACA阴性组(P<0.05),随着ACA转阴,肾功能逐渐恢复。结论ACA与狼疮性肾炎的肾功能损害密切相关,应用泼尼松和环磷酰胺降低血清ACA滴度,肾功能损害也随之好转。     

miR-16在肾癌中表达和临床意义的研究

目的检测肾癌组织及癌旁正常组织中miR-16的表达,分析miR-16表达水平与临床病理特征之间的关系,为深入研究miR-16在肾癌的发生发展中功能作用奠定基础。方法在标本库中随机挑选48例手术切除的肾癌及配对癌旁组织标本,提取总RNA并通过反转录获得cDNA,实时荧光定量PCR(qRT-PCR)检测标本中miR-16表达量,统计学分析其表达量与患者临床病理特征间的联系。结果 48对标本中41例(85.42%)肾癌标本miR-16表达上调,统计学分析证实肾癌组织中miR-16表达量明显高于配对的癌旁正常组织(P<0.01)。肾癌中miR-16表达量与患者性别、年龄、肾癌组织类型、TNM分期、AJCC临床分期无明显相关(P>0.05)。结论 miR-16在肾癌标本中明显表达上调,提示其可能与肾癌的发生发展有关。