Visualising E-selectin in the detection and evaluation of inflammatory bowel disease

Background—Vascular endothelial E-selectin expression is induced by proinflammatory cytokines andcontributes to accumulation of leucocytes in tissues.
Aims—To investigate the role ofE-selectin in inflammatory bowel disease (IBD).
Methods—E-selectin expression wasassessed in patients with ulcerative colitis and Crohn's disease bymeasuring the concentration of circulating soluble E-selectin(sE-selectin) using ELISA, by immunohistochemistry of colonic biopsyspecimens, and by abdominal immunoscintigraphy after injectingradiolabelled F(ab')₂ fragment of a monoclonalanti-E-selectin antibody. The value of scintigraphy usinganti-E-selectin was judged by a prospective comparative study ofautologous leucocyte scanning and E-selectin antibody scanning in 17 patients with IBD.
Results—Circulating sE-selectin waselevated in patients with clinically active disease. Tissue expressionof E-selectin was enhanced in patients with active inflammation, withweak or absent expression in inactive disease and healthy controls.In-111 labelled anti-E-selectin scintiscans were compared with Tc-99mlabelled leucocyte scans performed 24 hours earlier. Twelve patientshad areas of active inflammation on leucocyte scan while 11 patients had positive E-selectin scans. The results of the two scans were concordant in 14 patients, with those positive for both (10/17) showingsimilar disease localisation and extent.
Conclusions—Tissue E-selectinand circulating sE-selectin are increased during active inflammatorybowel disease. Anti-E-selectin imaging with radiolabelled monoclonalantibody identified areas of inflammation in Crohn's disease andulcerative colitis. The technique should prove useful clinically foridentifying the site and extent of disease.

Keywords:E-selectin; inflammatory bowel disease; Crohn'sdisease; ulcerative colitis

     

Ethnopharmacological in vitro studies on Austria's folk medicine—An unexplored lore in vitro anti-inflammatory activities of 71 Austrian traditional herbal drugs

Ethnopharmacological relevance

In Austria, like in most Western countries, knowledge about traditional medicinal plants is becoming scarce. Searching the literature concerning Austria's ethnomedicine reveals its scant scientific exploration.Aiming to substantiate the potential of medicinal plants traditionally used in Austria, 63 plant species or genera with claimed anti-inflammatory properties listed in the VOLKSMED database were assessed for their in vitro anti-inflammatory activity.

Material and methods

71 herbal drugs from 63 plant species or genera were extracted using solvents of varying polarities and subsequently depleted from the bulk constituents, chlorophylls and tannins to avoid possible interferences with the assays. The obtained 257 extracts were assessed for their in vitro anti-inflammatory activity. The expression of the inflammatory mediators E-selectin and interleukin-8 (IL-8), induced by the inflammatory stimuli tumor necrosis factor alpha (TNF-α) and the bacterial product lipopolysaccharide (LPS) was measured in endothelial cells. The potential of the extracts to activate the nuclear factors PPARα and PPARγ and to inhibit TNF-α-induced activation of the nuclear factor-kappa B (NF-κB) in HEK293 cells was determined by luciferase reporter gene assays.

Results

In total, extracts from 67 of the 71 assessed herbal drugs revealed anti-inflammatory activity in the applied in vitro test systems. Thereby, 30 could downregulate E-selectin or IL-8 gene expression, 28 were strong activators of PPARα or PPARγ (inducing activation of more than 2-fold at a concentration of 10 µg/mL) and 21 evoked a strong inhibition of NF-κB (inhibition of more than 80% at 10 µg/mL).

Conclusion

Our research supports the efficacy of herbal drugs reported in Austrian folk medicine used for ailments associated with inflammatory processes. Hence, an ethnopharmacological screening approach is a useful tool for the discovery of new drug leads.     

Genetic and Environmental Determinants of Variation of Soluble Adhesion Molecules

In our research we examined the contribution of putative genetic sources on interindividual variation and cross-sectional correlations of several adhesion molecules, including intracellular (ICAM-1) and vascular cell adhesion molecules (VCAM-1) and E-selectin, in a population-based sample of ethnically homogeneous families of European origin. The plasma levels of these molecules were measured in 947 apparently healthy individuals from 217 nuclear families. Quantitative statistical-genetic analysis implementing the model fitting technique revealed significant parent/offspring and sibling correlations (p < 0.01) for all three molecules. The putative genetic effects explained 55.2 ± 7.2% (VCAM-1), 63.3 ± 7.5% (ICAM) and 63.8 ± 8.1% (E-selectin) of the variation. Common family environmental factors also significantly influenced the variation of E-selectin (13%) and VCAM-1 (28.6%). The main results of our bivariate analysis showed that the observed phenotypic correlations between ICAM-1 and VCAM-1, and between ICAM-1 and E-selectin, were mostly attributable to shared environmental factors (  r_E= 0.896  and 0.737, respectively; p < 0.01). However, the correlation between VCAM-1 and E-selectin was likely caused by common genetic effects  (r_G= 0.334, p < 0.05)  . Our results show that familial clustering of adhesion molecules is likely due to strong genetic effects, supplemented with shared environmental factors.     

E-选择素在胚胎着床过程中小鼠子宫内膜的表达规律

目的：研究E-selectin在胚胎着床及早期分化中的作用。方法：用RT—PCR、Immunoblot、Western blot法检测小鼠胚胎着床前后蜕膜组织E-selectin的表达。结果：E-选择素在孕4天的小鼠子宫内膜中的表达出现升高，第5天达到峰值，随后妊娠第6、第7天E-selectin蛋白的表达则逐渐下降至正常水平。结论：提示E-选择素可能参与了胚胎着床过程。     

E-选择素预处理对大鼠脑缺血-再灌注损伤的影响

目的：研究E-选择素鼻粘膜耐受对大鼠脑缺血-再灌注损伤的作用及其机制。方法:E-选择素或PBS单程诱导耐受或加强诱导耐受48 h后,用改良的Zea Longa线栓法制备大鼠大脑中动脉缺血模型,缺血2 h再灌注22 h后,流式细胞仪测定血中CD3+CD4+T细胞及CD3+CD8+T细胞含量,TTC染色法测定脑梗死体积,RT-PCR检测脑梗死区E-选择素、细胞间黏附分子-1（ICAM-1）、淋巴细胞功能相关抗原-1（LFA-1）的表达,黄嘌呤氧化酶法检测脑组织中SOD水平。结果:E-选择素单程诱导耐受组CD3+CD4+T细胞与CD3+CD8+T细胞比值增加（P<0.05）。与其它组相比,E-选择素加强诱导耐受组脑梗死体积减小了40.87%（P<0.05）, CD3+CD8+T细胞所占比例减小、CD3+CD4+T细胞与CD3+CD8+T细胞比值增高（P<0.05）,脑组织中SOD水平升高(P<0.05),E-选择素、ICAM-1表达减少（P<0.05）,LFA-1表达有减少趋势。结论:E-选择素鼻黏膜耐受诱导脑缺血耐受可减轻脑缺血-再灌注损伤,其作用机制与CD8+ T细胞减少,CD4+T细胞与CD8+T细胞比值增加、SOD水平升高及E-选择素、ICAM-1表达减少有关。     

中枢神经系统感染与脑血管内皮细胞表面E选择素的表达

目的　研究中枢神经系统感染时血管内皮细胞E选择素的表达 ,以及地塞米松对其抑制作用。方法　将 5 4只小鼠随机分成侧脑室注射内毒素 (2 0 μg/2 0 μl)组、腹腔注射地塞米松 [1mg/(kg·12h) ]+侧脑室注射内毒素 (2 0 μg/2 0 μl)组、生理盐水组以及空白对照组。免疫组织化学法观察脑血管内皮细胞E选择素的表达 ,HE染色观察脑组织炎症反应。结果　空白对照组有少量E选择素表达 [(13 7± 2 7) %]。生理盐水组 [(13 5± 5 6 ) %]与空白对照组E选择素表达比较差异无显著性。内毒素组注射后E选择素表达在第 1天 [(31 8± 10 5 ) %]、第 2天 [(2 6 8± 9 6 ) %]明显增多 ,与生理盐水组比较差异有显著性 (P <0 0 1或 <0 0 5 ) ,注射后第 3天 [(15 7± 9 9) %]恢复正常 (P >0 0 5 ) ;注射后 3d内E选择素表达呈进行性下降 (F =4 0 8,P <0 0 5 )。地塞米松 +内毒素组于注射后第 1天和第 2天E选择素表达与内毒素组比较明显受到抑制 ,分别为 (13 0± 12 4 ) %、(8 8±6 0 ) %(P略 >0 0 5 ,P <0 0 5 ) ,与生理盐水组比较差异无显著性 ;同时脑组织炎症反应受到明显抑制。结论　中枢神经系统感染时脑血管内皮细胞E选择素的表达增加 ;地塞米松可通过抑制脑血管内皮细胞表面E选择素的表达来抑制炎症反应     

Comparative study of adhesion molecule expression in cultured human macro- and microvascular endothelial cells

Culture systems as models for disease are only valid as long as they are comparable to in vivo conditions. The phenotype of cultured endothelial cells (ECs) has only been sporadically compared to the corresponding phenotype in vivo. Thus, we compared by immunolocalization the endothelial expression of ICAM-1, VCAM, and E-selectin in vivo in stimulated/unstimulated human umbilical vein endothelial cells (HUVEC) as a model for macrovascular ECs and stimulated/unstimulated HPMEC (human pulmonary microvessel endothelial cells) as a model for pulmonary microvascular ECs with that in human lungs in vivo (normal and ARDS). Proinflammatory stimuli in vitro were used to stimulate conditions relevant for ARDS. ICAM-1 expression in stimulated HUVEC/HPMEC correlated well with in vivo expression (macro- and microvessels). For E-selectin, the staining pattern in macro/microvessels correlated moderately with unstimulated and well with stimulated HUVEC/HPMEC. For VCAM a good correlation was found for stimulated/unstimulated HUVEC and unstimulated HPMEC. The expression patterns in stimulated HUVEC corresponded well for all three molecules with those in vivo. Thus, the expression patterns in vitro are only partially transferable to in vivo conditions. The study suggests that E-selectin- and VCAM-coated beads could potentially serve in the isolation process of arteriolar and venular ECs.     

Does infection with Chlamydia pneumoniae and/or Helicobacter pylori increase the expression of endothelial cell adhesion molecules in humans?

Objective   To investigate if Chlamydia pneumoniae and/or Helicobacter pylori seropositivity is associated with elevated levels of soluble endothelial cell adhesion molecules (sCAMs) as markers of atherosclerotic activity.
Methods   Immunoglobulin A (IgA) and IgG antibodies to the two bacteria, soluble intercellular cell adhesion molecule-1 (sICAM-1), soluble vascular cell adhesion molecule-1 (sVCAM-1) and E-selectin were measured in coronary heart disease (CHD) patients ( n  = 193) and age- and sex-matched controls ( n  = 193). Two different serological methods were used for the detection of Chlamydia antibodies: Labsystems microimmunofluorescence to detect species-specific C. pneumoniae antibodies and Medac's recombinant enzyme-linked immunosorbent assay to detect genus-specific lipopolysaccharide antibodies.
Results   The concentrations of sICAM-1 and E-selectin were higher in CHD patients with positive vs. negative Chlamydia lipopolysaccharide IgA ( P  = 0.044 for both). H. pylori antibodies alone did not predict raised levels of sCAMs, but in CHD patients sICAM-1 was increased with IgA seropositivity to both bacteria compared to double seronegativity ( P  = 0.034). Concentrations of sVCAM-1 were elevated in CHD patients with double IgA seropositivity compared to those with Chlamydia lipopolysaccharide IgA seropositivity alone ( P  = 0.018).
Conclusion   Our results may indicate that C. pneumoniae contributes to increased inflammation in CHD, and that this contribution is even more pronounced when present in combination with H. pylori IgA antibodies.     

E-选择素、CD14及VCAM1水平与生殖道沙眼衣原体感染的相关性分析

目的 探讨E-选择素、CD14及血管细胞粘附分子(vascular cell adhesion molecule 1,VCAM1)水平与生殖道沙眼衣原体感染的相关性.方法 选取86例生殖道沙眼衣原体感染患者为观察组;根据生殖道沙眼衣原体感染患者是否伴有炎症,分为感染伴炎症组和感染无炎症组,各43例;同时,选取36例健康体检者为对照组;采用双抗体夹心酶联免疫吸附法(ELISA)测定所有研究对象的血清E-选择素、CD14及VCAM1水平,并作对比分析;观察组患者采取炎克宁冲剂结合阿奇霉素治疗,对比治疗前后的血清E-选择素、CD14及VCAM1水平,并与对照组作对比分析.结果 观察组的血清E-选择素、CD14及VCAM1水平均显著高于对照组;差异具有统计学意义(P＜0.05);感染伴炎症组与感染无炎症组的血清E-选择素、CD14及VCAM1水平的差异具有统计学意义(P＜0.05);观察组患者采取炎克宁冲剂结合阿奇霉素治疗后,临床总有效率为94.19％,生殖道沙眼衣原体转阴率为87.21％;治疗后的血清E-选择素、CD14及VCAM1水平显著低于治疗前水平,差异具有统计学意义(P＜0.05);与对照组对比,差异无统计学意义(P＞0.05).结论 生殖道沙眼衣原体感染的病情进展与E-选择素、CD14及VCAM1水平密切相关,通过监测E-选择素、CD14及VCAM1水平,可为生殖道沙眼衣原体感染的治疗、评估疗效及预后而提供依据.