高密度脂蛋白中的1-磷酸鞘氨醇对心肌细胞生存信号通路的影响

目的:观察高密度脂蛋白(HDL)不同组分中1-磷酸鞘氨醇(S1P)的含量对心肌细胞AKT和ERK1/2信号转导通路的影响,探讨HDL对心肌细胞的保护作用。方法:应用超速离心法和层析法,分离不同组分HDL;应用地高辛核素标记法检测各组分中S1P含量。给予小鼠心肌细胞不同S1P含量的HDL组分和S1P1、S1P3受体抑制剂VPC23019刺激后,用蛋白印迹法检测AKT和ERK1/2转导通路磷酸化水平的变化。结果 :与对照组比较,受不同HDL组分刺激后,成年小鼠心肌细胞AKT和ERK1/2磷酸化水平升高,且随着各组分中S1P含量升高,磷酸化水平呈上升趋势,而S1P1及S1P 3受体抑制剂VPC23019能明显阻断此种作用。结论:不同HDL组分通过细胞膜上的S1P受体,激活PI3K-AKT和MEK-ERK1/2信号通路,该作用通过S1P1和S1P3受体介导;HDL可能通过S1P发挥心肌保护作用。     

鞘氨醇-1-磷酸酯受体拮抗剂对抗肾小球基底膜肾炎的影响

目的观察鞘氨醇-1-磷酸酯受体拮抗剂720(sphingosine-1-phosphate receptor agonist 720,FTY720)对C57BL/6小鼠抗肾小球基底膜肾炎模型(Anti-GBM GN)的干预作用并对其作用机制进行探讨。方法选择健康雄性C57BL/6小鼠52只,给予兔IgG(500μg)加完全弗氏佐剂皮下注射进行预免疫,5d后从中随机抽取8只作为正常对照组,经尾静脉注射同剂量的非抗体性的正常兔血清,其余44只小鼠取灭活兔抗小鼠GBM抗血清0.3mL/20g经尾静脉注射。44只小鼠按随机化原则分组,FTY0.3mg/(kg·d)组(n=8)尾静脉注射6h后0.3mg/(kg·d)FTY720灌胃;FTY3mg/(kg·d)组(n=8)3mg/(kg·d)FTY720灌胃;FTY5mg/(kg·d)组(n=8)5mg/(kg·d)FTY720灌胃;FTY10mg/(kg·d)组(n=8)10mg/(kg·d)FTY720灌胃;Anti-GBM GN组(n=12)为模型组,同等剂量的无菌生理盐水灌胃。观察小鼠实验室指标及肾脏组织病理学改变情况。实时定量PCR检测小鼠脾脏鞘氨醇-1-磷酸脂(sphingosine-1-phosphate,S1P)受体S1P1～S1P5mRNA表达变化。流式细胞仪检测CD4+T细胞的凋亡情况。结果治疗组小鼠的尿蛋白、血肌酐、尿素氮和血胆固醇水平明显降低,血浆白蛋白明显增高;治疗组小鼠肾脏组织学病变明显减轻。与模型组相比,用药各组随着用药剂量增加S1P1、S1P2、S1P5的表达量逐渐减少,差异有显著性;S1P3、S1P4的表达量无明显变化,差异无显著性。流式细胞仪检测结果表明FTY720促进小鼠脾脏CD4+T细胞凋亡。结论 FTY720可能通过下调S1P1、S1P2、S1P5mRNA表达,促进CD4+T细胞凋亡,减轻抗肾小球基底膜肾炎模型的肾脏组织学改变,从而减少蛋白尿、稳定肾功能。     

Modulation of sphingosine-1-phosphate in ulcerative colitis

ABSTRACT

Introduction: Sphingosine-1-phosphate (S1P) is a membrane-derived lysophospholipid signaling molecule implicated in various physiological and pathological processes, such as regulation of the immune, cardiovascular, pulmonary, and nervous systems and theoretical cancer-related risks, through extracellular activation of S1P1-5 receptors.

Areas covered: S1P receptor agonism is a novel strategy for the treatment of UC targeting lymphocyte recirculation, through blockade of lymphocyte egress from lymph nodes. We conducted an extensive literature review on PUBMED on currently available data on molecular aspects of S1P modulation, the mechanisms of action of S1PR agonists (fingolimod, ozanimod, etrasimod, and KRP-203), and their potential efficacy and safety for the treatment of patients with ulcerative colitis.

Expert opinion: Selective S1P modulators have emerged to enlarge the efficacy and safety profile of this class of agents. Phase 3 programs should add the potential body of evidence to prove their benefit for the management of UC patients.     

1-磷酸鞘氨醇对血小板贮存损伤的影响

目的探讨1-磷酸鞘氨醇对手工浓缩血小板的贮存损伤(活化率、凋亡率和细胞内游离Ca2+)的影响。方法向手工浓缩血小板中分别加入0.1、0.5和1.0μmol/L 3个浓度的S1P,采用流式细胞术检测血小板22℃保存2、3、5、7 d的活化率、凋亡率和细胞内游离Ca2+。结果 0.5和1.0μmol/L的外源性S1P可以降低血小板内游离Ca2+的浓度,降低其活化率和凋亡率。结论 1-磷酸鞘氨醇对血小板的贮存损伤起到一定的抑制作用。     

血清葡萄糖-6-磷酸异构酶与自身免疫性疾病相关性研究

目的探讨血清葡萄糖-6-磷酸异构酶(GPI)水平与各种自身免疫性疾病(AID)的关系。方法比较正常对照和各种AID确诊病例的血清GPI浓度,分析GPI在各种AID中的检测价值。结果类风湿性关节炎(RA)组的GPI浓度和阳性率都显著高于其他AID组和正常对照组(P<0.001),系统性红斑狼疮(SLE)组、干燥综合征(SS)组、多发性肌炎(PM)组、进行性系统性硬皮病(SSC)组和强直性脊柱炎(AS)组GPI的阳性率均在25%以下,显著低于RA组的阳性率(71.0%)。结论在各种AID中,RA组血清中GPI升高最明显。血清GPI检测可作为RA的诊断指标。如果GPI和类风湿因子(RF)联合检测,将对RA的诊断具有较高价值。     

鞘氨醇-1-磷酸在脓毒症中调节作用的研究进展

鞘氨醇-1-磷酸(sphingosine-1-phosphate,S1P)是具有多种生物活性的鞘磷脂代谢产物,通过激活细胞膜表面的G蛋白偶联受体即S1P受体(sphingosine-1-phosphate receptors,S1PRs),参与细胞生长和凋亡、免疫与凝血系统调节等多种生理功能。脓毒症是由于感染引起免疫反应失调所致的严重危及生命的疾病,常导致多器官功能障碍甚至衰竭。脓毒症导致的器官衰竭主要与内皮细胞和免疫细胞在炎症环境中的病理生理变化有关,包括血管通透性增加、血栓形成、炎症及免疫反应失调。S1P可参与调节脓毒症的多种病理生理过程,有望成为预测脓毒症患者病情严重程度的重要标志物,也是治疗脓毒症的潜在靶点。本文通过对S1P在脓毒症发生发展过程中的调节作用及相关临床研究进行总结,旨在为该领域的后续研究提供参考。     

S1PR3激动剂KRX-725促进细菌清除影响脓毒症小鼠预后

目的 通过合成特异性S1PR3激动剂KRX-725,探讨S1PR3参与细菌清除的功能及其分子机制.方法 20只健康清洁级雄性C57BL/6小鼠以随机数字法分为S1PR3特异性激动剂KRX-725治疗组和对照组.采用大肠杆菌(3×106)腹腔注射诱导脓毒症模型,治疗组小鼠于模型后立即腹腔注射KRX-725(10 mg/kg),对照组给予等体积的溶剂,比较两组小鼠48 h生存率(n=12).模型后24 h检测腹腔及血液细菌负荷(n=5),并取肺组织进行苏木精-伊红染色(HE染色)评估KRX-725治疗组及对照组的肺脏病理损伤程度(n=3).体外采用大肠杆菌刺激腹腔巨噬细胞(细胞∶细菌=1∶10),分别加入KRX-725及其对照溶剂,采用CM-H2 DCFDA探针标记细胞内活性氧,酶标仪检测检测不同时间点巨噬细胞活性氧水平.庆大霉素保护实验观察KRX-725对巨噬细胞的清除细菌能力的影响.生存率分析采用Log-rank test,组间数据比较采用独立样本t检验分析,P＜0.05为差异有统计学意义.结果 KRX-725治疗组脓毒症小鼠48 h生存率较对照组显著提高(P＜0.05).脓毒症模型后24 h,KRX-725治疗组较对照组血液及腹腔灌洗液中的细菌负荷显著降低(血液:t=3.17,P＜0.05;腹腔灌洗液:t=4.07,P＜0.01),同时KRX-725可显著降低脓毒症模型肺组织损伤程度(肺损伤评分:KRX-725组1.4 ±0.25,对照组2.4 ±0.25)(t=2.89,P＜0.05).体外实验,KRX-725治疗组较对照组可迅速上调大肠杆菌刺激后20 min及30 min细胞内活性氧的水平(20 min荧光强度:KRX-725组522.9±38.76,对照组385.9±15.90,P＜0.05;30min荧光强度:KRX-725组519.7 ±25.02,对照组384.5±15.28,P＜0.01).庆大霉素保护实验发现,KRX-725可显著降低细菌吞噬后3 h及6h巨噬细胞内存活的大肠杆菌的数量(3 h:KRX-725组286.5±98.35,对照组710.8 ±--107.8,P＜0.05;6 h:KRX-725组72.5±6.45,对照组205.8±66.76,P＜0.01).结论 体内应用KRX-725可改善脓毒症小鼠生存率,降低血液及腹腔细菌负荷,减轻肺脏损伤,从而改善脓毒症预后.体外KRX-725通过上调巨噬细胞活性氧的水平,提高巨噬细胞对清除细菌的能力.     

Transactivation of sphingosine-1-phosphate receptors by FcepsilonRI triggering is required for normal mast cell degranulation and chemotaxis

Mast cells secrete various substances that initiate and perpetuate allergic responses. Cross-linking of the high-affinity receptor for IgE (FcepsilonRI) in RBL-2H3 and bone marrow-derived mast cells activates sphingosine kinase (SphK), which leads to generation and secretion of the potent sphingolipid mediator, sphingosine-1-phosphate (S1P). In turn, S1P activates its receptors S1P1 and S1P2 that are present in mast cells. Moreover, inhibition of SphK blocks FcepsilonRI-mediated internalization of these receptors and markedly reduces degranulation and chemotaxis. Although transactivation of S1P1 and Gi signaling are important for cytoskeletal rearrangements and migration of mast cells toward antigen, they are dispensable for FcepsilonRI-triggered degranulation. However, S1P2, whose expression is up-regulated by FcepsilonRI cross-linking, was required for degranulation and inhibited migration toward antigen. Together, our results suggest that activation of SphKs and consequently S1PRs by FcepsilonRI triggering plays a crucial role in mast cell functions and might be involved in the movement of mast cells to sites of inflammation.     

类风湿关节炎患者关节液血红素氧合酶-1水平的变化及临床意义

目的 检测类风湿关节炎(RA)患者关节液中血红素氧合酶-1(HO-1)水平,探讨HO-1在RA中的临床意义.方法 对50例RA患者(活动期24例,非活动期26例)及30名正常对照者,采用酶联免疫吸附试验(ELISA)检测HO-1、基质金属蛋白酶-3(MMP-3)水平和葡萄糖6-磷酸异构酶(GPI)在关节液中的含量.结果...     

血清1-磷酸鞘氨醇、生长分化因子-15、尿酸水平对急性呼吸窘迫综合征患者临床预后影响

目的 探讨血清1-磷酸鞘氨醇(S1P)、生长分化因子-15(GDF-15)、尿酸水平对急性呼吸窘迫综合征(ARDS)患者临床预后的影响,分析其用于预测患者预后的价值.方法 选取自2018年1月至2020年1月收治的100例ARDS患者为观察组,根据氧合指数将患者分为轻度组(200 mmHg<氧合指数≤300 mmHg,...     

MMP-9及其抑制剂TIMP-1与多发性硬化活动相关性研究

目的探讨血清和脑脊液中基质金属蛋白酶（ MMP）-9及其抑制剂金属蛋白酶组织抑制剂（ TIMP）-1与多发性硬化发病及疾病活动的相关性。方法本实验共分四组：多发性硬化组、神经系统炎性疾病（OIND）组、神经系统非炎性疾病（NIND）组和健康人群组。采用酶联免疫吸附试验（ELISA）检测血清MMP-9、血清和脑脊液中TIMP-1的表达水平;采用酶谱法检测脑脊液中MMP-9水平;以MRI检查为多发性硬化疾病活动性判断标准,分析比较多发性硬化活动组和非活动组MMP-9及TIMP-1水平。结果多发性硬化组血清MMP-9水平显著高于NIND组（ P＜0.05）和健康人群组（ P＜0.001）,血清TIMP-1水平显著低于OIND组（ P＜0.001）、NIND组（ P＜0.001）和健康人群组（ P＜0.001）;多发性硬化组脑脊液中MMP-9水平高于NIND组（ P＜0.05）,脑脊液中TIMP-1水平显著低于OIND组（ P＜0.05）;多发性硬化活动期患者血清MMP-9水平、MMP-9/TIMP-1比值分别显著高于非活动期（ P均＜0.05）,活动期血清TIMP-1水平与非活动期无统计学差异。结论检测血清和脑脊液中MMP-9及TIMP-1水平对多发性硬化疾病活动性的监测有一定参考价值。     

Elevated cell-associated levels of interleukin 1β and interleukin 6 in inflamed mucosa of inflammatory bowel disease

The aim of this study was to investigate the involvement of the monocyte-derived cytokines interleukin 1β (IL-1β), interleukin 6 (IL-6) and tumour necrosis factor-α (TNF-α) in idiopathic inflammatory bowel disease. Endoscopic biopsies of normal and inflamed intestinal mucosa were obtained from patients with ulcerative colitis ( n  = 11) and with Crohn's disease ( n  = 10). Intestinal mucosal cells were isolated by collagenase digestion. Cell viability, morphology and CD14 expression were determined. To measure cell-associated cytokine levels, cells were lysed and analysed for IL-1β and TNF-α in specific radioimmunoassays and for IL-6 using a biological assay. Compared with mucosal cells from control patients without inflammatory bowel disease the inflamed intestine in ulcerative colitis and Crohn's disease displayed markedly enhanced levels of IL-1β (median 245 pg 10⁻⁶ cells, range 30–1275) and IL-6 (median 22 U 10⁻⁶ cells, range 1–298). Non-inflamed mucosa in patients with ulcerative colitis and Crohn's disease did not shown elevated levels of IL-1β (median 50 pg 10⁻⁶ cells, range 33–90) or IL-6 (mean below detection limit of assay, i.e. 1 U 10⁻⁶ cells). In contrast, no clear cut difference between inflamed and non-inflamed mucosa could be detected for TNF-α. High tissue levels of IL-6 were associated with a high endoscopic grade of local inflammation. These results suggest that the monocyte-derived cytokines IL-1β and IL-6 are mediators of inflammation in inflammatory bowel disease.     

Diminished neurokinin-1 receptor availability in patients with two forms of chronic visceral pain

Central sensitization and dysregulation of peripheral substance P and neurokinin-1 receptor (NK-1R) signaling are associated with chronic abdominal pain in inflammatory bowel disease (IBD) and irritable bowel syndrome (IBS). Although positron emission tomography (PET) has demonstrated that patients with injury-related chronic pain have diminished NK-1R availability in the brain, it is unknown whether these deficits are present in IBD and IBS patients, who have etiologically distinct forms of non-injury-related chronic pain. This study’s aim was to determine if patients with IBD or IBS exhibit deficits in brain expression of NK-1Rs relative to healthy controls (HCs), the extent to which expression patterns differ across patient populations, and if these patterns differentially relate to clinical parameters. PET with [¹⁸F]SPA-RQ was used to measure NK-1R availability by quantifying binding potential (BP) in the 3 groups. Exploratory correlation analyses were performed to detect associations between NK-1R BP and physical symptoms. Compared to HCs, IBD patients had NK-1R BP deficits across a widespread network of cortical and subcortical regions. IBS patients had similar, but less pronounced deficits. BP in a subset of these regions was robustly related to discrete clinical parameters in each patient population. Widespread deficits in NK-1R BP occur in IBD and, to a lesser extent, IBS; however, discrete clinical parameters relate to NK-1R BP in each patient population. This suggests that potential pharmacological interventions that target NK-1R signaling may be most effective for treating distinct symptoms in IBD and IBS.     

肿瘤坏死因子样配体1A在炎症性肠病免疫调控中的作用

炎症性肠病包括溃疡性结肠炎和克罗恩病,目前多认为其发病机制是环境因素作用于遗传易感者,在肠道菌群的参与下,启动了难以停止的、发作与缓解交替的肠道免疫反应。然而,其发病机制尚不十分明确,因此,进一步探讨和阐明炎症性肠病的发病机制对于寻找新的治疗方法具有重要的意义。肿瘤坏死因子样配体1A(tumor necrosis factor-like ligand 1 aberrance,TL1A)是新近发现的一种肿瘤坏死因子家族新成员,是炎症性肠病的易感基因。TL1A在黏膜炎症的固有免疫和适应性免疫之间起到了关键的调控和连接作用,从而在炎症性肠病中发挥重要作用。现将TL1A在炎症性肠病免疫调控机制中的作用进行综述。     

Mucosa‐associated lymphoid tissue lymphoma translocation protein 1 in rheumatoid arthritis: Longitudinal change after treatment and correlation with treatment efficacy of tumor necrosis factor inhibitors

BackgroundMucosa‐associated lymphoid tissue lymphoma translocation protein 1 (MALT1) correlates with treatment outcomes in inflammatory bowel disease and rheumatoid arthritis (RA). This study aimed to further evaluate the MALT1 longitudinal change and its relationship with tumor necrosis factor inhibitors (TNFi) response in RA patients.MethodsSeventy‐one RA patients receiving TNFi [etanercept (n = 42) or adalimumab (n = 29)] were enrolled. MALT1 was detected by RT‐qPCR in peripheral blood samples of RA patients before treatment (W0), at week (W)4, W12, and W24 after treatment. RA patients were divided into response/non‐response, remission/non‐remission patients according to their treatment outcome at W24. Meanwhile, MALT1 was also detected by RT‐qPCR in 30 osteoarthritis patients and 30 healthy controls (HCs).ResultsMucosa‐associated lymphoid tissue lymphoma translocation protein 1 was elevated in RA patients compared with HCs (Z=−6.392, p < 0.001) and osteoarthritis patients (Z = −5.020, p < 0.001). In RA patients, MALT1 was positively correlated with C‐reactive protein (r_s  = 0.347, p = 0.003), but not other clinical characteristics, treatment history, or current TNFi category. Meanwhile, MALT1 decreased from W0 to W12 in total RA patients (x²  = 86.455, p < 0.001), etanercept subgroup (x²  = 46.636, p < 0.001), and adalimumab subgroup (x²  = 41.291, p < 0.001). Moreover, MALT1 at W24 (p = 0.012) was decreased in response patients compared with non‐response patients; MALT1 at W12 (p = 0.027) and W24 (p = 0.010) were reduced in remission patients than non‐remission patients. In etanercept subgroup, MALT1 at W24 (p = 0.013) was decreased in response patients compared with non‐response patients. In adalimumab subgroup, MALT1 at W24 (p = 0.015) was lower in remission patients than non‐remission patients.ConclusionMucosa‐associated lymphoid tissue lymphoma translocation protein 1 reduction after treatment is associated with response and remission to TNFi in RA patients.     

Incidence,Prevalence, Costs,and Impact on Disability of Common Conditions Requiring Rehabilitation in the United States: Stroke,Spinal Cord Injury,Traumatic Brain Injury,Multiple Sclerosis,Osteoarthritis, Rheumatoid Arthritis,Limb Loss,and Back Pain

Objective

To determine the relative incidence, prevalence, costs, and impact on disability of 8 common conditions treated by rehabilitation professionals.

Data Sources

Comprehensive bibliographic searches using MEDLINE, Google Scholar, and UpToDate, (June, 2013).

Data Extraction

Two review authors independently screened the search results and performed data extraction. Eighty-two articles were identified that had relevant data on the following conditions: Stroke, Spinal Cord Injury, Traumatic Brain Injury, Multiple Sclerosis, Osteoarthritis, Rheumatoid Arthritis, Limb Loss, and Back Pain.

Data Synthesis

Back pain and arthritis (osteoarthritis, rheumatoid arthritis) are the most common and costly conditions we analyzed, affecting more than 100 million individuals and costing greater than $200 billion per year. Traumatic brain injury, while less common than arthritis and back pain, carries enormous per capita direct and indirect costs, mostly because of the young age of those involved and the severe disability that it may cause. Finally, stroke, which is often listed as the most common cause of disability, is likely second to both arthritis and back pain in its impact on functional limitations.

Conclusions

Of the common rehabilitation diagnoses we studied, musculoskeletal conditions such as back pain and arthritis likely have the most impact on the health care system because of their high prevalence and impact on disability.