神经导向因子Netrin-1调节单核巨噬细胞迁移与动脉粥样斑块关系的研究进展

动脉粥样硬化是动脉壁的一种慢性炎症性疾病,单核巨噬细胞在其发生发展中起着关键作用。动脉粥样斑块中单核巨噬细胞迁移能力受损,滞留于斑块内,增加了斑块不稳定性,加速动脉粥样硬化病变的进展。目前研究表明动脉粥样斑块中巨噬细胞分泌的神经导向因子Netrin-1通过与巨噬细胞表面相应受体结合,可以抑制巨噬细胞迁出斑块,促进动脉粥样硬化的进展。但在动脉粥样硬化形成初期,血管内皮细胞表达的Netrin-1却被发现对动脉粥样硬化起到保护作用。     

Targeted IL‐4 therapy synergizes with dexamethasone to induce a state of tolerance by promoting Treg cells and macrophages in mice with arthritis

F8‐IL‐4 is a recently developed immunocytokine that delivers IL‐4 to sites of inflammation by targeting the neovasculature. We previously reported that F8‐IL‐4, in combination with dexamethasone (DXM), provides a durable therapy in mice with collagen‐induced arthritis (CIA). Therefore, the objective of this study was to identify the mechanism by which IL‐4 and DXM combination therapy provides long‐lasting disease remission. F8‐IL‐4 alone attenuated inflammation in CIA and this was associated with increased T_H2 and decreased T_H17 cell numbers in the joints. Similarly, DXM alone had an antiinflammatory effect associated with lower T_H17 cell numbers. In both cases, these therapeutic benefits were reversed once treatment was stopped. On the other hand, combination therapy with F8‐IL‐4 plus DXM led to a synergistic increase in the percentage of regulatory T (Treg) cells and antiinflammatory macrophages in the arthritic joint and spleen as well as IL‐10 levels in serum and spleen. The net result of this was a more pronounced attenuation of inflammation and, more importantly, protection from arthritis relapse post therapy retraction. In conclusion, F8‐IL‐4 plus DXM is a durable treatment for arthritis that acts by promoting Treg cells in a synergistic manner, and by producing a sustained increase in antiinflammatory macrophages.     

人参提取液对小鼠巨噬细胞的辐射防护作用

本文利用图象分析系统(TAS)定量细胞化学和环核苷酸免疫荧光细胞化学技术观察了人参提取液对小鼠巨噬细胞的辐射防护作用。定量测定表明人参可明显提高受照小鼠巨噬细胞内ANAE、AcPase、ATPase和PAS阳性物质的含量。同时,环核苷酸免疫细胞化学亦显示受照小鼠施加人参后巨噬细胞cAMP和cGMP免疫荧光的定位和强度发生变化,恢复至正常定位和强度。本文对结果的意义进行了讨论     

单个巨噬细胞超氧阴离子水平的定量检测方法

目的：建立单个巨噬细胞超氧阴离子（O₂^-.）水平的定量检测方法,从而达到在单细胞内同时检测O₂^-.和细胞内游离钙离子浓度（[Ca²⁺]_i）的目的。方法：分离小鼠腹腔巨噬细胞，用NBT还原法显示细胞产生O₂^-.的水平，以细胞加入NBT前与染色后的透光强度之比来定量表示该细胞产生O₂^-.的量；以荧光探针Fura-2/AM负载细胞检测单细胞内[Ca²⁺]_i。结果：①分别用PMA、OZ刺激细胞并用NBT还原法染色后细胞内均有黑色沉淀，但前者分布均匀而后者所产生沉淀多偏于一侧，呈明显的局域性分布；②用NBT孵育10 min后，细胞透光强度没有明显变化（n=43, P>0.05），用NBT刺激细胞产生黑色沉淀后细胞透光强度明显降低（P<0.01, n=43）。③在不同的聚焦状态下，无细胞背景区的透光强度没有明显变化，而细胞区透光强度变化明显，但各个细胞透光强度的变化趋势是一致的；④PMA处理3批巨噬细胞并用NBT法染色得到3组R_OI值的分布情况提示用R_OI值定量是合理的但批间差异过大；⑤静息状态下的小鼠腹腔巨噬细胞[Ca²⁺]_i与细胞受PMA刺激后的R_OI呈显著正相关（n=43, r=0.42, P<0.01）。结论：用NBT还原法染色前后细胞透光强度比值可以代表该细胞产生O₂^-.的量，进而我们可以在单细胞内同时检测O₂^-.和[Ca²⁺]_i。     

去甲肾上腺素对大鼠腹腔巨噬细胞内IP_3、［Ca~（2＋）］_i和Ia抗原表达

本文应用细胞膜放射标记和离子交换层析法测定巨噬细胞（ＭФ）内肌醇－１，４，５－三磷酸（ＩＰ３）、用Ｃａ￣（２＋）指示剂的分光光谱法测定ＭФ内Ｃａ￣（２＋）浓度（［Ｃａ￣（２＋）］）、用ＡＰＡＡＰ桥联酶标法检测ＭФ表面Ｉａ抗原的表达，研究去甲肾上腺素（ＮＥ）对大鼠腹腔的ＭФ内ＩＰ３、［Ｃａ￣（２＋）］ｉ和ＭФ表面ｌａ抗原表达的影响。结果显示ＮＥ（１０￣（－８）ｍｏｌ／Ｌ）可显著增高ＭФ中ＩＰ＿３含量（４４２±２２ｃｐｍ／１０￣６ｃｅｌｌｓ，对照组１０２±８ｃｐｍ／１０￣６ｃｅｌｌｓ，Ｐ＜０．０１）；ＮＥ可使ＭФ内［Ｃａ￣（２＋）］ｉ显著增高（３２２±７８ｎｍｏｌ／Ｌ，对照组９７±１７ｎｍｏｌ／Ｌ，Ｐ＜０．０１）；ＮＥ可显著增高ＭФ表面Ｉ－Ａ、Ｉ－Ｅ抗原的表达（６４±８％、５８±６％，对照组５０±３％，４４±４％，Ｐ＜０．０１）。提示神经递质ＮＥ促进ＭФ表面Ｉａ抗原表达的作用可能是通过第二信使ＩＰ＿３和Ｃａ￣（２＋）介导的。     

Microarray analysis of gene expression profiles in response to treatment with bee venom in lipopolysaccharide activated RAW 264.7 cells

AIM OF THE STUDY: The therapeutic application of bee venom (BV) has been used in traditional medicine to treat diseases such as arthritis, rheumatism and pain. Macrophages produce molecules that are known to play roles in inflammatory responses. MATERIAL AND METHODS: We performed microarray analysis to evaluate the global gene expression profiles of RAW264.7 macrophage cells treated with BV. In addition, six genes were subjected to real-time PCR to confirm the results of the microarray. The cells were treated with lipopolysaccharide (LPS) or BV plus LPS for 30 min or 1h. RESULTS: 124 genes were found to be up-regulated and 158 were found to be down-regulated in cells that were treated with BV plus LPS for 30 min, whereas 211 genes were up-regulated and 129 were down-regulated in cells that were treated with BV plus LPS for 1h when compared with cells that were treated with LPS alone. Furthermore, the results of real-time PCR were similar to those of the microarray. BV inhibited the expression of specific inflammatory genes that were up-regulated by nuclear factor-kappa B in the presence of LPS, including mitogen-activated protein kinase kinase kinase 8 (MAP3K8), TNF, TNF-alpha-induced protein 3 (TNFAIP3), suppressor of cytokine signaling 3 (SOCS3), TNF receptor-associated factor 1 (TRAF1), JUN, and CREB binding protein (CBP). CONCLUSIONS: These results demonstrate the potent activity of BV as a modulator of the LPS-mediated nuclear factor-kappaB (NF-kappaB)/MAPK pathway in activated macrophages. In addition, these results can be used to understand other effects of BV treatment.     

Differential flow cytometric detection of intracellular cytokines in peripheral and peritoneal mononuclear cells of women with endometriosis

OBJECTIVE: The pathogenesis of endometriosis is related to functional changes in CD3+ and CD14+ cells observed both at the local and systemic level. Here we investigated whether, and if so, how the body compartment influences cytokine expression in stimulated peritoneal and peripheral CD3+ and CD14+ cells of women with endometriosis. STUDY DESIGN: Isolated peripheral blood (PB) and peritoneal fluid (PF) mononuclear cells from women with endometriosis were cultured under non-adherent conditions and stimulated with PMA and ionomycin for 6h to induce intracellular cytokine synthesis of TNF-alpha, IFN-gamma, and IL-8 by CD3+ cells or with LPS for 9h to produce TNF-alpha, IL-6, IL-10, MCP-1, and IL-8 by CD14+ cells. RESULTS: The percentages of positive CD3+ cells stained for TNF-alpha and IFN-gamma were significantly higher and those stained for IL-8 were significantly lower in PF compared with PB, this being independent of the stage of endometriosis. In contrast, the percentages of CD14+ cells producing TNF-alpha, IL-6, IL-10, MCP-1, and IL-8 were significantly higher in PB than PF of women with endometriosis. CONCLUSIONS: Monocytes/macrophages and lymphocytes derived from the peripheral and peritoneal compartments of women with endometriosis differentially respond to stimulated cytokine synthesis induction. However, it is difficult to state whether the observed phenomenon is more related to body compartment influence per se or to the presence of endometriosis.     

高通量基因芯片初步筛选ITP脾和正常脾巨噬细胞的差异表达基因

目的研究免疫性血小板减少性紫癜(ITP)患者脾和正常脾巨噬细胞的差异表达基因。方法提取9例ITP患者及9例正常人脾巨噬细胞的总RNA,反转录制备含荧光分子Cy5标记的cDNA探针,与含有30968点cDNA的表达谱芯片杂交后扫描荧光强度,筛选出差异表达的基因并进行分析。结果基因芯片共筛选出1545个差异表达基因,其中上调基因718个,下调基因827个。差异表达基因涉及免疫反应、细胞黏附及受体调节、细胞信号转导、细胞骨架和运动代谢、细胞凋亡、酶调活性等方面。差异表达基因生物路径涉及Toll样受体信号通路,Fcγ受体介导吞噬通路,促分裂原活化蛋白激酶信号通路、细胞内吞通路等。结论基因芯片筛选ITP患者脾巨噬细胞的差异表达基因是有效的。差异表达基因可能为ITP发病机制的研究提供新证据和治疗靶标。