褪黑素受体激动剂改善高糖高脂喂养大鼠脂联素敏感性

目的 观察褪黑素受体激动剂(NEU-P11)对高糖高脂饲养大鼠脂联素敏感性的影响.方法 将30 只SD大鼠随机分为对照组(CD组),高糖高脂组(HFSD组),褪黑素组(Mel组),褪黑素受体激动剂组(NEU-P11组).CD组饲以正常饲料;其余3组饲以高糖高脂饲料.6个月后,给药治疗2个月.治疗期间,Mel组每天注射Mel(4 mg/kg);NEU-P11组每天注射NEU-P11(10 mg/kg);CD组以及HFSD组注射生理盐水(5 ml/kg).测定糖脂代谢指标并做口服葡萄糖耐量实验(oral glucose tolerant test,OGTT),Western印迹检测脂联素(adiponectin,APN)在脂肪组织及脂联素受体(AdipoR)在骨骼肌组织中的表达变化.结果 高糖高脂饮食可诱导SD大鼠产生胰岛素抵抗,脂联素表达增加.Neu-P11治疗后,胰岛素敏感性增强,脂联素表达降低至正常水平.结论 Neu-P11能提高胰岛素敏感性,改善脂联素抵抗.     

脂氧素受体及其配体在炎症反应中作用的研究进展

炎症反应参与人体多种疾病的发生发展。脂氧素受体(FPR2/ALX)通过结合不同的配体,在炎症相关疾病中,参与炎症的调节和疾病的进展过程。淀粉样蛋白A、β淀粉样蛋白(Aβ42)和一些微生物源性分子结合FPR2/ALX后促进炎症的发生发展;内源性脂类介质脂氧素和消退素D激活FPR2/ALX,产生抗炎、促炎症消退作用。FPR2/ALX在炎症反应中表现出生物功能的多样性,为新药研究和炎性疾病的治疗提供作用靶点和研究方向。本文主要综述了FPR2/ALX与其配体在炎症反应过程中的作用。     

脂氧素受体激动剂BML-111对脊髓缺血再灌注损伤模型大鼠的神经保护作用

BACKGROUND: The mechanisms of spinal cord ischemia-reperfusion injury are the result of the combined effects of multiple factors, but there is no effective treatment.     

NF-kB介导脂氧素A4拮抗TNF-α所致的系膜细胞白介素的合成

为研究脂氧素A4(LXA4)拈抗肿瘤坏步E因子α(TNF-α)对肾小球系膜细胞的白介素(IL）-β(IL-1β)、IL-6合成的作用。对体外培养大鼠肾小球系膜细胞，用不同浓度的LXA4预刺激，再加入TNF-α共同孵育；或单用TNF-α刺激肾小球系膜细胞。在孵育后用ELISA法检测培养上清中的IL-1β,IL-6蛋白表达量；用RT-PCR法检测IL-1β、IL-6的mRNA表达量。应用凝胶电泳迁移率试验（EMSA)测定核因子-kB(NF-KB)的DNA结合活性。结果发现，LXA4呈剂量依赖性地抑制TNF-α诱导的肾小球系膜细胞IL-1β和IL-6蛋白的合成与mRNA表达，抑制NF-kB的DNA结合活性。说明LXA4通过下调NF-kB的DNA结合活性，拮抗TNF-α对肾小球系膜细胞的IL-1β、IL-6合成的促进作用。     

脂氧素A4拮抗肿瘤坏死因子α对系膜细胞Jak1/STAT3途径的活化

目的：验证脂氧素A4(LXA4) 是否抑制肿瘤坏死因子α(TNFα) 所致的大鼠肾小球系膜细胞的增殖，并探讨其作用中信号转导的分子机制。 方法： 对体外培养的大鼠肾小球系膜细胞，用不同浓度的LXA4 预刺激，再加入TNFα共同孵育，或单用TNFα刺激系膜细胞。用MTT渗入法检测细胞的增殖。用凝胶电泳迁移率试验（EMSA）检测信号转导子和转录激活子-3（STAT3）的活性。用RT-PCR法检测细胞周期素E的mRNA表达。用Western blotting法检测细胞周期素E的蛋白表达量。结果： LXA4呈剂量依赖性地抑制TNFα诱导的肾小球系膜细胞的增殖、STAT3结合活性增加、细胞周期素E mRNA表达与蛋白合成的亢进。结论： LXA4能够抑制TNFα所致的大鼠系膜细胞的增殖，其机制可能是阻断Jak1/STAT3信号转导途径。     

巨细胞病毒感染与急性冠状动脉综合征患者血清sP-选择素和肿瘤坏死因子的变化

目的　探讨人巨细胞病毒(HCMV)感染与急性冠状动脉综合征(ACS)患者血清sP 选择素、肿瘤坏死因子(TNF α)的变化及相关性。方法　采用酶联免疫吸附技术检测79例ACS患者、30例稳定性心绞痛(SA)患者和30例正常对照组血清HCMV IgM、HCMV IgG、sP 选择素和TNF α的水平。结果　(1)在ACS、SA及正常对照组中,血清HCMV IgM、IgG的阳性率分别为30 4 % (2 4 79)、10 0 %(3 30 )和6 7% (2 30 ) ;86 1% (6 8 79)、80 0 % (2 4 30 )和5 3 3% (16 30 )。HCMV IgM的阳性率在ACS组中明显高于SA组和对照组(P <0 0 1) ,HCMV IgG的阳性率在ACS和SA组中明显高于对照组(P <0 0 1)。(2 )与SA组和对照组相比,ACS组中血清sP 选择素和TNF α水平显著升高,分别为(15 2 0 0±112 7)和(14 81 0±10 9 1)pg ml比(6 4 37 3±6 6 6 9)pg ml,(2 7 3±13 7)pg ml和(2 8 1±11 3)pg ml比(5 6 2±18 4 )pg ml,组间差异有统计学意义(P <0 0 1)。随着冠状动脉(冠脉)病变程度的增加,ACS中急性心肌梗塞(AMI)组与不稳定性心绞痛(UA)组相比,sP 选择素和TNF α水平显著升高(P <0 0 1) ,而SA组与对照组间差异无统计学意义(P >0 0 5 )。(3)ACS组中HCMV IgM阳性患者血清中sP 选择素和TNF α水平较HCMV IgM阴性患者明显升高(P <0 0     

脂氧素抑制脂多糖诱导的单核巨噬细胞向树突状样细胞分化

目的：脂氧素是天然免疫／炎症反应中的重要“刹车信号”，具有促进炎症及时缓解的独特功能；但其对特异性免疫应答有何调节作用尚缺乏直接证据，为此，本研究观察了脂氧素对脂多糖诱导的单核巨噬细胞向树突状样细胞分化过程的影响。方法：体外培养小鼠巨噬细胞RAW264．7细胞，脂多糖（1mg／L）处理24h诱导其分化为树突状样细胞，实验分为空白组、脂多糖组及脂氧素处理组（1mg／L脂多糖与100nmol脂氧素共同孵育）；各组处理24h后Giemsa染色高倍镜下计数树突状样细胞比例，流式细胞仪检测细胞表面共刺激分子CDS0（B7-1）、CD86（B7-2）的表达水平。Westem blot检测NF-κB核转位及I-κB降解情况。     

人巨细胞病毒感染THP-1源性巨噬细胞对IL-1β表达的影响

目的 观察HCMV感染THP-1源性巨噬细胞后,促炎细胞因子IL-1β表达的时序性变化.方法 构建HCMV感染THP-1源性巨噬细胞模型,设立HCMV AD169标准毒株感染组、模拟感染对照组、LPS+ATP对照组和poly(dA:dT)对照组.用ELISA法测定THP-1源性巨噬细胞培养上清IL-1β水平在病毒感染细胞后lh、3h、6h、12h、24 h和48 h的时序性变化.分别用real-time PCR和western blot检测感染后6 h IL-1β基因和蛋白的表达水平.结果 HCMV感染THP-1源性巨噬细胞6h后,促炎细胞因子IL-1β基因的相对表达量是模拟感染组的7.77倍.HCMV感染THP-1源性巨噬细胞1h后,细胞上清IL-1β表达量逐渐显著增高,感染后3h和6h继续升高,感染后12h达到高峰,一直持续到48 h.HCMV感染THP-1源性巨噬细胞6h HCMV感染组IL-1β蛋白表达明显高于模拟感染对照组和poly(dA:dT)对照组,而与LPS+ATP对照组比较无统计学差异.结论 HCMV感染THP-1源性巨噬细胞可诱导IL-1β高表达,且呈时序性增高趋势.     

NF-κB介导脂氧素A4拮抗TNF-α所致的系膜细胞白介素的合成

为研究脂氧素A4(LXA4)拮抗肿瘤坏死因子α(TNF-α)对肾小球系膜细胞的白介素(IL)-1β(IL-1β)、IL-6合成的作用。对体外培养大鼠肾小球系膜细胞,用不同浓度的LXA4预刺激,再加入TNF-α共同孵育;或单用TNF-α刺激肾小球系膜细胞。在孵育后用ELISA法检测培养上清中的IL-1β、IL-6蛋白表达量;用RT-PCR法检测IL-1β、IL-6的mRNA表达量。应用凝胶电泳迁移率试验(EMSA)测定核因子-κB(NF-κB)的DNA结合活性。结果发现,LXA4呈剂量依赖性地抑制TNF-α诱导的肾小球系膜细胞IL-1β和IL-6蛋白的合成与mRNA表达,抑制NF-κB的DNA结合活性。说明LXA4通过下调NF-κB的DNA结合活性,拮抗TNF-α对肾小球系膜细胞的IL-1β、IL-6合成的促进作用。     

脂氧素对脂多糖诱导的巨噬细胞游离钙浓度变化及活性氧的影响

近年来越来越多的研究资料表明炎症反应的及时消退是炎症治疗的重要环节。脂氧素对多种炎症细胞和炎症相关基因有显著的负性调节作用，是体内重要的内源性脂质促炎症消退介质。细胞内游离钙离子浓度变化为巨噬细胞活化的重要第二信使。但脂氧素对巨噬细胞内游离钙浓度变化了解较少。为此，本研究采用细胞内钙离子特异性荧光探针Fluo3-AM和活性氧特异性荧光探针DCFH-DA同时标记小鼠巨噬细胞，用激光共聚焦显微镜同时动态观察脂多糖引起巨噬细胞胞内游离钙浓度和活性氧变化及脂氧素的影响作用。     

Induction of soluble tumor necrosis factor receptor (sTNF-R75) release by HIV adsorption on cultured human monocytes

Soluble tumor necrosis factor (TNF) receptors were recently detected in the circulation of patients with early HIV-induced disease, at significantly higher levels than in control subjects. They were proposed as markers of disease progression and of the degree of immunodeficiency. We report that adsorption of heat-inactivated HIV-1 LAI to isolated human monocytes triggers the release of both TNF-α and its natural specific inhibitor, the soluble TNF receptor (sTNF-R)75, but not that of sTNF-R55. Only limited inhibition of sTNF-R release was obtained in the presence of a fully neutralizing anti-TNF-α monoclonal antibody, suggesting that stimulation by TNF-α was only partially responsible for sTNF-R release. HIV-1 LAI induced a higher sTNF-R/TNF ratio than lipopolysaccharide, a well-known monocyte activator. Monocytes thus represent a cellular source of sTNF-R that can be detected in the circulation of HIV-infected patients from seroconversion onwards. The release of sTNF-R could be of great significance in the control of HIV infection via the cytokine network and especially TNF-α.     

Mechanisms of human cytomegalovirus infection with a focus on epidermal growth factor receptor interactions

Human cytomegalovirus (HCMV) is a widespread opportunistic herpesvirus that causes severe diseases in immunocompromised individuals. It has a high prevalence worldwide that is linked with socioeconomic factors. Similar to other herpesviruses, HCMV has the ability to establish lifelong persistence and latent infection following primary exposure. HCMV infects a broad range of cell types. This broad tropism suggests that it may use multiple receptors for host cell entry. The identification of receptors used by HCMV is essential for understanding viral pathogenesis, because these receptors mediate the early events necessary for infection. Many cell surface components have been identified as virus receptors, such as epidermal growth factor receptor (EGFR), which is characterized by tyrosine kinase activity and plays a crucial role in the control of key cellular transduction pathways. EGFR is essential for HCMV binding, signaling, and host cell entry. This review focuses on HCMV infection via EGFR on different cell types and its implications for the cellular environment, viral persistence, and infection.     

Differential effect of Fc gamma receptor ligation on LPS-stimulated TNF-alpha secretion by hepatic,splenic, and peritoneal macrophages

Lipopolysaccharide (LPS) increases serum TNF- levels due to TNF- secretion by macrophages. The serum TNF- response to LPS was augmented 10× when FcR ligation was induced by the intravenous injection of Gig-coated erythrocytes (IgG) prior to the administration of LPS. The macrophage population responsible for the augmented TNF- secretion was determined by isolating Kupffer cells, splenic macrophages and peritoneal macrophages from mice that had been given ElgG prior to LPS and determining TNF- secretion ex vivo. The intravenous injection of ElgG augmented LPS-stimulate TNF- secretion by Kupffer cells and splenic macrophages. In contrast, LPS-stimulated TNF- secretion by peritoneal macrophages was not altered by either the intravenous or intraperitoneal injection of ElgG. In vitro phagocytosis of ElgG by isolated peritoneal macrophages also did not augment LPS-stimulated TNF- secretion. These results show that FcR ligation augments LPS-stimulated TNF- secretion by Kupffer cells and splenic macrophages but not by peritoneal macrophages. Thus, the ability of FcR ligation to influence TNF- secretion may be specific to the tissue source of the macrophages.     

人工瓣膜置换体外循环过程中痰热清注射液的肺保护

背景：体外循环心内直视术不可避免会造成肺损伤,近年来临床上已有中医药应用于该领域减轻肺损伤的研究。 目的：观察痰热清注射液对机械瓣置换患者体外循环肺损伤的保护效果。 方法：心脏瓣膜置换患者40例随机分为2组,痰热清组患者于术前晚、体外循环前分别予以痰热清注射液20 mL入250 mL生理盐水静脉滴注,对照组予以生理盐水250 mL静滴。于体外循环前、体外循环40 min、体外循环停机、停机后2,6,24 h共6个不同时间点抽取桡动脉血2 mL,全血细胞分析仪测定中性粒细胞数量;双抗体夹心ELISA法测定可溶性细胞间黏附因子1、白细胞介素8血浆浓度。并于体外循环前、体外循环停机时取左右心房血测中性粒细胞并计算跨肺差值。于瓣膜置换切皮前、体外循环结束后10 min、瓣膜置换结束时、瓣膜置换结束后4 h 4个时相计算两组患者的呼吸指数与肺动态顺应性。 结果与结论：两组患者体外循环后各个时间点中性粒细胞值、可溶性细胞黏附因子1、白细胞介素8较体外循环前明显升高(P < 0.01),痰热清组明显低于对照组(P < 0.01)。体外循环停机时中性粒细胞跨肺差值明显高于体外循环前(P < 0.01),痰热清组明显低于对照组(P < 0.01)。两组呼吸指数较瓣膜置换切皮前明显升高(P < 0.01),肺动态顺应性较体外循环前明显降低(P < 0.01),瓣膜置换结束后4 h恢复至切皮前水平(P > 0.05),痰热清组呼吸指数较对照组降低(P < 0.05),肺动态顺应性瓣膜置换结束时较对照组升高(P < 0.01)。提示痰热清注射液能减轻机械瓣置换后的肺损伤,有较好的肺保护效果。     

Two inhibitors of pro-inflammatory cytokine release,interleukin-10 and interleukin-4, have contrasting effects on release of soluble p75 tumor necrosis factor receptor by cultured monocytes

The biological activity of the pro-inflammatory cytokine, tumor necrosis factor (TNF)-α depends on the level of TNF-α itself, the expression of the p55 and p75 cell surface receptors for TNF on target cells and the concentrations of the natural inhibitors of TNF-α, the soluble p55 and p75 TNF receptors (TNF-R). Interleukin (IL)-10 and IL-4 are known to inhibit TNF-α production by monocytes. We, therefore, investigated the effects of IL-10 and IL-4 on the cell surface expression and release of TNF-R by human monocytes to determine whether these cytokines also indirectly modulated the biological activity of TNF-α. Exposure to IL-10 (1-10 U/ml) for 24 or 48 h increased soluble p75 TNF-R expression and concomitantly reduced surface expression of p75 TNF-R. Further, IL-l α-stimulated production of TNF-α was diminished by IL-10 and only a small proportion of this TNF-α was bioactive, consistent with increased production of inhibitory soluble TNF-R. IL-10 also induced down-regulation of surface p55 TNF-R on monocytes, and increased release of soluble p55 TNF-R. However, the expression of soluble p55 TNF-R was much lower than soluble p75 TNF-R, indicating that it contributed less importantly to neutralization of TNF-α under these conditions. Like IL-10, IL-4 supressed the release of TNF-α by monocytes. In contrast to IL-10, however, IL-4 (0.1-10 ng/ml) supressed the release of soluble p75 TNF-R from monocytes in a dose-dependent manner. Release of soluble p55 TNF-R was also supressed by IL-4. IL-10, therefore, reduces the pro-inflammatory potential of TNF in three ways: by down-regulating surface TNF-R expression whilst increasing production of soluble TNF-R and inhibiting the release of TNF-α itself. This suggests that IL-10 may be useful in the treatment of diseases where overexpression of TNF-α occurs.     

An mRNA atlas of G protein-coupled receptor expression during primary human monocyte/macrophage differentiation and lipopolysaccharide-mediated activation identifies targetable candidate regulators of inflammation

G protein-coupled receptors (GPCRs) are among the most important targets in drug discovery. In this study, we used TaqMan Low Density Arrays to profile the full GPCR repertoire of primary human macrophages differentiated from monocytes using either colony stimulating factor-1 (CSF-1/M-CSF) (CSF-1 M?) or granulocyte macrophage colony stimulating factor (GM-CSF) (GM-CSF M?). The overall trend was a downregulation of GPCRs during monocyte to macrophage differentiation, but a core set of 10 genes (e.g. LGR4, MRGPRF and GPR143) encoding seven transmembrane proteins were upregulated, irrespective of the differentiating agent used. Several of these upregulated GPCRs have not previously been studied in the context of macrophage biology and/or inflammation. As expected, CSF-1 M? and GM-CSF M? exhibited differential inflammatory cytokine profiles in response to the Toll-like Receptor (TLR)4 agonist lipopolysaccharide (LPS). Moreover, 15 GPCRs were differentially expressed between these cell populations in the basal state. For example, EDG1 was expressed at elevated levels in CSF-1 M? versus GM-CSF M?, whereas the reverse was true for EDG6. 101 GPCRs showed differential regulation over an LPS time course, with 65 of these profiles being impacted by the basal differentiation state (e.g. GPRC5A, GPRC5B). Only 14 LPS-regulated GPCRs showed asynchronous behavior (divergent LPS regulation) with respect to differentiation status. Thus, the differentiation state primarily affects the magnitude of LPS-regulated expression, rather than causing major reprogramming of GPCR gene expression profiles. Several GPCRs showing differential profiles between CSF-1 M? and GM-CSF M? (e.g. P2RY8, GPR92, EMR3) have not been widely investigated in macrophage biology and inflammation. Strikingly, several closely related GPCRs displayed completely opposing patterns of regulation during differentiation and/or activation (e.g. EDG1 versus EDG6, LGR4 versus LGR7, GPRC5A versus GPRC5B). We propose that selective regulation of GPCR5A and GPCR5B in CSF-1 M? contributes to skewing toward the M2 macrophage phenotype. Our analysis of the GPCR repertoire expressed during primary human monocyte to macrophage differentiation and TLR4-mediated activation provides a valuable new platform for conducting future functional analyses of individual GPCRs in human macrophage inflammatory pathways.     

High polymeric IgA content facilitates recognition of microbial polysaccharide-natural serum antibody immune complexes by immobilized human galectin-1

While interleukin (IL)-33, a novel member of the IL-1 family, seems to promote T helper type 2 (Th2)-associated inflammations and allergic diseases, the stimulating factors for IL-33 production are less well characterized. Prostaglandin E₂ (PGE₂) has been shown to suppress immune cell functions. However, the immune enhancement by this mediator is not well understood. In the present study, we examined the effect of PGE₂ on IL-33 production by dendritic cells (DCs). Bone marrow-derived DCs were stimulated with lipopolysaccharide (LPS) in the presence or absence of PGE₂. LPS increased mRNA expression of the IL-1 family members, IL-1, IL-18, and IL-33 in DCs. PGE₂ alone showed slight effect on IL-1, IL-18, and IL-33 mRNA expression in DCs. Of note, LPS combined with PGE₂ caused in a synergistic enhancement of mRNA expression of IL-33 but not IL-1 and IL-18. In addition, PGE₂ dramatically enhanced IL-33 protein production by DCs upon LPS stimulation. The protein kinase A (PKA) inhibitor H89 significantly inhibited the PGE₂-mediated enhancement of IL-33 production by DCs. Thus, PGE₂ appears to enhance IL-33 mRNA expression and its protein synthesis via PKA pathway in DCs. PGE₂ may promote Th2-mediated inflammations through the enhancement of IL-33 production by DCs, which might be associated with the pathogenesis of allergic diseases.     

Effects of blood sample handling procedures on measurable inflammatory markers in plasma, serum and dried blood spot samples

The interests in monitoring inflammation by immunoassay determination of blood inflammatory markers call for information on the stability of these markers in relation to the handling of blood samples. The increasing use of stored biobank samples for such ventures that may have been collected and stored for other purposes, justifies the study hereof. Blood samples were stored for 0, 4, 24, and 48 h at 4 degrees C, room temperature (RT), and at 35 degrees C, respectively, before they were separated into serum or plasma and frozen. Dried blood spot samples (DBSS) were stored for 0, 1, 2, 3, 7, and 30 days at the same temperatures. 27 inflammatory markers in serum and plasma and 25 markers in DBSS were measured by a previously validated multiplex sandwich immunoassay using Luminex xMAP technology. The measurable concentrations of several cytokines in serum and plasma were significantly increased when blood samples were stored for a period of time before the centrifugation, for certain cytokines more than 1000 fold compared to serum and plasma isolated and frozen immediately after venepuncture. The concentrations in serum generally increased more than in plasma. The measurable concentrations of inflammatory markers also changed in DBSS stored under various conditions compared to controls frozen immediately after preparation, but to a much lesser degree than in plasma or serum. The study demonstrates that trustworthy measurement of several inflammatory markers relies on handling of whole blood samples at low temperatures and rapid isolation of plasma and serum. Effects of different handling procedures for all markers studied are given. DBSS proved to be a robust and convenient way to handle samples for immunoassay analysis of inflammatory markers in whole blood.