辐射对巨噬细胞极化和功能的调节作用

巨噬细胞是存在于所有组织中的调节性细胞,它们对辐射刺激的反应机制复杂,既有共同的应对感染、损伤的变化过程,也有自己独特的极化转变方式。辐射相关的极化转变决定了巨噬细胞的功能类型。巨噬细胞受到不同的照射后,受到不同细胞因子的调节,涉及多个信号通路,极化程度也有所不同。本文综述辐射对巨噬细胞极化和功能的调节作用,以期对免疫基础研究和临床放射治疗提供参考。     

大鼠创伤后腹腔巨噬细胞亚群的变化及与IL-6、TNFα的关系

目的 探讨创伤后免疫功能紊乱状态与巨噬细胞亚群的关系。方法 采用创伤失血模型，运用流式细胞仪分析单克隆抗体ED1、ED2单克隆抗体双标记表达情况，测定腹腔巨噬细胞（pMΦ）培养上清白细胞介素－6（IL－6）、肿瘤坏死因子（TNFα）含量（ELISA法）。结果 正常大鼠pMΦ分为EDI^ ED2^-亚群、ED1^ ED2^ 亚群，无单纯的ED2^ 巨噬细胞亚群，创伤后ED1^ ED2^-亚群比例增加，ED1^ ED2^ 亚群比例降低，同时TNFα、IL－6分泌增加，内毒素（LPS）刺激可增加TNFα、IL－6分泌水平。结论 创伤后细胞因子分泌紊乱可能与ED1^ ED2^-巨噬细胞亚群有关。     

肺泡巨噬细胞与急性肺损伤

肺泡巨噬细胞是肺部防御病原微生物和肺损伤的第一道防线 ,是目前受到重视的肺巨噬细胞亚群之一 ,对急性肺损伤的发生、发展及转归具有重要影响。本文着重综述近年来肺泡巨噬细胞在急性肺损伤中的作用的研究进展     

运动与代谢性炎症反应——有氧运动改善机体代谢机制研究进展

最新研究认为,机体脂肪组织除了储存能量之外,还是分泌多种脂肪因子的重要内分泌器官。脂肪组织功能失调常伴发肥胖、胰岛素抵抗等代谢性疾病,其原因可能与机体代谢性炎症反应有关。机体在肥胖状态下血液中的单核巨噬细胞进入组织内分化为M1型促炎巨噬细胞,M1型巨噬细胞可释放多种细胞因子并作用于体内胰岛素敏感细胞,抑制细胞胰岛素信号敏感性。同时,血液中淋巴细胞、嗜酸性粒细胞和中性粒细胞等也可通过影响巨噬细胞的迁移及活化而发挥代谢调节作用。因此,巨噬细胞对于发展组织代谢性炎症及维持代谢状态至关重要。本综述总结目前国内外有关运动改变机体组织内多种转录调控因子从而影响巨噬细胞极化及代谢性炎症调控的研究进展,为揭示运动改善机体代谢状态的机制提供理论依据。     

大鼠严重胸部创伤肺泡与间质巨噬细胞分泌TNF-α、IL-6的差异及意义

目的 观察大鼠严重胸部创伤后肺泡（AM）及间质（IM）巨噬细胞两类亚群分泌TNF—α、IL-6的篇异,并探讨其意义。方法 利用小型多功能生物撞击机,以400kPa驱动压力对大鼠右侧上胸壁进行致伤,建立大鼠严重胸部创伤模型,支气管肺泡灌洗,机械结合酶消化法分离、培养肺泡及间质巨噬细胞,动态检测创伤前、创伤后2、4、8、16、24小时以及复合LPS攻击后肺泡及间质巨噬细胞分泌TNF—α、IL-6的水平。结果 创伤复合LPS攻击后,AM,IM分泌TNF-α、IL-6增加,伤后4、8、16小时时相点AM分泌TNF—α显著高于IM;伤后每个时相点IM分泌IL-6均显著高于AM。结论 大鼠严重胸部创伤对AM、IM分泌TNF—α、IL-6具有不同影响,其在创伤后机体免疫功能紊乱的过程中具有不同作用,本研究为创伤性急性肺损伤的发病机制提供了一定的实验及理论依据。     

P388D_1类巨噬细胞系Fc受体的观察

本文通过测定P388D_1细胞的吞噬作用和Fc受体,观察其免疫功能.它具有巨噬细胞相似的吞噬功能.可吞噬经兔抗鸡红细胞(CRBC)抗体调理的CRBC,吞噬率和吞噬指数受到抗体稀释度的影响.在稀释度1∶1500时,吞噬率和吞噬指数分别为90%、4.3.经EA玫瑰花环试验证明P388D_1细胞膜表面存有Fc受体,它的花环形成率为90%.这说明它是一个较均一的细胞群.并可用于巨噬细胞在体外免疫功能研究中作为替代物.     

Tim-3对巨噬细胞功能的调节作用研究

目的探讨Tim-3对巨噬细胞功能的调节作用及其机制。方法通过RNA干扰降低Tim-3在巨噬细胞表面的表达,构建一种稳定低表达Tim-3的RAW264.7巨噬细胞系;以野生型及低表达Tim-3的巨噬细胞为研究对象,探讨Tim-3对巨噬细胞活性及表型的影响。结果将含Tim-3 siRNA及NC对照序列的载体经脂质体分别转染细胞系RAW264.7,以G418加压筛选,获得低表达Tim-3的巨噬细胞稳定株。功能实验显示,激活Tim-3通路促进了巨噬细胞分泌TNF-α、IL-6。同时发现Tim-3通路可以影响CD80/CD86共刺激分子在巨噬细胞表面的表达,并进而影响巨噬细胞的抗原呈递功能,促进Th1、Th17细胞的极化。结论成功构建了重组载体Tim3 siRNA,建立了稳定低表达Tim-3的巨噬细胞系,初步研究结果提示Tim-3信号通路在促进TNF-α、IL-6分泌及调节巨噬细胞共刺激分子表达,促进Th1、Th17细胞分化中发挥重要作用。     

游泳训练对小鼠巨噬细胞功能影响的实验研究

对游泳训练3个月的C_(57)B L/6小鼠巨噬细胞功能进行了研究。结果每日运动30分钟组巨噬细胞吞噬功能和抑瘤活性增强,运动60分钟组巨噬细胞IL-1分泌水平和吞噬功能增高,运动120分钟组的巨噬细胞IL-1分泌水平和吞噬功能则明显降低。提示不同运动量的运动训练引起巨噬细胞功能变化有差异,适量运动可以增强巨噬细胞功能,过大的运动则对巨噬细胞功能有损害或抑制作用。     

脑缺血炎症反应的活体内MR监测

脑缺血后的炎性反应过程中有大量的巨噬细胞，超小型顺磁性氧化铁粒子（USPIO）是能够被网状内皮系统清除的一种新型MR对比剂，可以作为巨噬细胞示踪剂。吞噬USPIO的巨噬细胞在T1WI上呈高信号，T2WI上表现为低信号。综述USPIO及其在脑缺血炎性反应监测中的作用。     

炎症在内支架植入术后再狭窄中的作用及地塞米松干预的实验研究

目的 观察活化巨噬细胞培养上清液对原代培养的大鼠胸主动脉平滑肌细胞（VSMCs）增殖的影响，以及地塞米松的干预作用。材料与方法 体外培养大鼠VSMCs并制备活化巨噬细胞培养上清液，利用四甲基偶氮唑盐（MTT）法及^3H-胸腺嘧啶脱氧核苷（^3H-TdR）掺人法检测不同浓度的活化巨噬细胞上清液对VSMCs增殖的效果，并用地塞米松进行干预，观察地塞米松对巨噬细胞活化及其对VSMCs增殖的影响。结果 （1）活化巨噬细胞培养上清液促进VSMCs的增殖，呈浓度依赖性，随浓度增加，吸光度（OD）值及每分钟闪烁次数（cpm）值也相应增加，方差分析，值分别为15．23和23．98（P〈0．01）；两两比较各组间差异显著（P〈0．05）；（2）不同浓度地塞米松干预后OD值及cpm值随地塞米松浓度增加而降低，方差分析F值分别为63．36及13．07（P〈0．01）；两两比较10^-11mol／L、10^-9moL／L及对照组之间无统计学差异，其余各组间差异显著。结论 活化巨噬细胞促进VSMCs增殖，表明炎症反应是血管内支架植入术后再狭窄的重要机制之一。地塞米松能够抑制其促增殖作用，可能在一定程度上达到预防或治疗再狭窄的作用。     

Effects of ALA-PDT on the macrophages in wound healing and its related mechanisms in vivo and in vitro

BackgroundSeveral studies have suggested the effectiveness of photodynamic therapy (PDT) for wound healing. Macrophages are critical immune cells necessary for regulated inflammation during wound repair. However, the available information regarding the effects of PDT on macrophages during cutaneous wound healing remains insufficient. This study aimed to further investigate these aspects in vivo and in vitro.MethodsMouse full-thickness wound models were used as the study samples to investigate the therapeutic effects and mechanisms of 5-aminolevulinic acid (ALA) PDT. Wound healing rate, granulation tissue formation, local inflammation, M1/M2 macrophages differentiation, were measured at different time points treated by ALA-PDT. The polarization of macrophages induced by ALA-PDT was further evaluated in vitro using PCR and western blot analysis.ResultsALA-PDT could promote formation of granulation tissue, increase inflammatory infiltration and activate M1 macrophages in the early stage of injury. While, ALA-PDT could also facilitate absorption of granulation tissue, inhibit inflammatory infiltration and enhance M2 macrophages polarization in the later stage of wound repair. In vitro, ALA-PDT could modulate the ratio of M2 polarization to M1 polarization via NF-κB signaling pathway.ConclusionsALA-PDT topical application stimulates wound healing by regulating formation of granulation tissue, inflammatory process and M1/M2 macrophages differentiation. The study places a preliminary theoretical basis for topical ALA-PDT to be administered clinically in cutaneous wounds healing.     

Low dose ionising radiation leads to a NF-κB dependent decreased secretion of active IL-1β by activated macrophages with a discontinuous dose-dependency

Abstract Purpose: Therapy with low doses of ionising radiation (X-rays) exerts anti-inflammatory effects. Little is known about whether and how low doses of X-ray treatment modulate the inflammatory phenotype of macrophages, especially the secretion of Interleukin-1beta (IL-1β). Materials and methods: Macrophages were differentiated from human THP-1 monocytes, activated with lipopolysaccharide (LPS), treated with distinct low doses of X-rays, and co-activated with monosodium urate crystals (MSU) to induce inflammasome activation. Secretion of IL-1β was analysed by an enzyme-linked immunosorbent assay (ELISA) and Western blot. Furthermore, we analysed the intracellular amounts of the serine/threonine protein kinase B (named: Akt), mitogen-activated protein kinase p38 (p38), the v-rel reticuloendotheliosis viral oncogene homolog A (RelA), and pro- and cleaved IL-1β. Results: Low dose X-rays led to decreased secretion of active IL-1β in a manner discontinuous with dose which was most pronounced after 0.5 or 0.7 Gy. Passive release of lactate dehydrogenase (LDH) was not influenced by X-rays. The decreased secretion of IL-1β correlated with reduced translocation of RelA, being part of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) complex, into the nucleus. After 0.5 or 0.7 Gy of X-rays, the intracellular protein amounts of up (p38) and downstream molecules (Akt) of NF-κB were reduced in activated macrophages, as were the pro- and cleaved forms of IL-1β. Conclusions: Distinct low doses of X-rays induce an anti-inflammatory phenotype of activated macrophages by lowering the amount of secreted IL-1β in a NF-κB dependent manner.     

肿瘤相关巨噬细胞MR成像可行性的实验研究

目的 探讨肿瘤相关巨噬细胞(TAMs)MR成像的可行性.方法 BALB/c小鼠和BALB/c(nu/nu)裸鼠按数字表法随机分组,每组3只,小鼠分为6个组(分别为注射前组,注射后6、24、48、72 h、7 d组)和裸鼠5个组(分别为注射前,注射后6、24、48、72 h).用含20μg Fe/ml人血清白蛋白和多巴胺包裹的铁氧颗粒(HAS-IONPs)标记巨噬细胞系(RAW 264.7)24 h,收集细胞,经尾静脉注射5×106RAW 264.7至皮下4T1荷瘤小鼠和22B移植瘤裸鼠,分别在上述不同时间点应用7.0 T MR扫描仪行横断面、冠状面T2WI,动物处死后行病理检查.细胞毒性试验应用SPSS 11.5软件行单因素方差分析;MPd信号强度采用ImageJ 1.42软件测量.结果 普鲁士蓝染色及电镜证实细胞标记有效,单个细胞吸入的铁量为6.19 Pg.HAS-IONPs孵育细胞24 h后,不同浓度HAS-IONPs (5、10、20、40、80、160μg/ml)每组细胞平均吸光度A值分别为1.95 ±0.19、1.82±0.29、2.10±0.14、1.96±0.18、2.05±0.27、2.17±0.22,与对照组(2.00±0.07)相比,细胞吸光度A值差异无统计学意义(F=1.24,P>0.05).标记HAS-IONPs细胞数与MR R2值呈线性正相关,r=0.99,P<0.05.4T1肿瘤:注射后6 h T2WI坏死囊变周围呈明显低信号,信号强度下降率为59.4%,肿瘤实质区信号未见明显改变,下降率为4.8%.注射后24 h坏死与肿瘤实质交界部位见不规则环形低信号影,下降率为46.8%,肿瘤坏死囊变周围T:WI信号恢复,恢复96.8%.22B肿瘤:注射后6 h MRI见多发散在灶性低信号影,下降率为64.3%,24 h见点片状低信号影缩小.2种肿瘤注射后48、72 h或7 d MRT2WI形态、位置和注射后24 h相似,但信号强度递减.病理与影像结果一致.结论 TAMs MR成像可行.注射后24 h T2WI是显示肿瘤巨噬细胞迁移、浸润及定位肿瘤内的最佳时机,注射后6 h可能反映病变局部血流灌注及新生血管的状态,不同肿瘤巨噬细胞分布不同.     

Low dose ionising radiation leads to a NF-κB dependent decreased secretion of active IL-1β by activated macrophages with a discontinuous dose-dependency

Abstract

Purpose: Therapy with low doses of ionising radiation (X-rays) exerts anti-inflammatory effects. Little is known about whether and how low doses of X-ray treatment modulate the inflammatory phenotype of macrophages, especially the secretion of Interleukin-1beta (IL-1β).

Materials and methods: Macrophages were differentiated from human THP-1 monocytes, activated with lipopolysaccharide (LPS), treated with distinct low doses of X-rays, and co-activated with monosodium urate crystals (MSU) to induce inflammasome activation. Secretion of IL-1β was analysed by an enzyme-linked immunosorbent assay (ELISA) and Western blot. Furthermore, we analysed the intracellular amounts of the serine/threonine protein kinase B (named: Akt), mitogen-activated protein kinase p38 (p38), the v-rel reticuloendotheliosis viral oncogene homolog A (RelA), and pro- and cleaved IL-1β.

Results: Low dose X-rays led to decreased secretion of active IL-1β in a manner discontinuous with dose which was most pronounced after 0.5 or 0.7 Gy. Passive release of lactate dehydrogenase (LDH) was not influenced by X-rays. The decreased secretion of IL-1β correlated with reduced translocation of RelA, being part of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) complex, into the nucleus. After 0.5 or 0.7 Gy of X-rays, the intracellular protein amounts of up (p38) and downstream molecules (Akt) of NF-κB were reduced in activated macrophages, as were the pro- and cleaved forms of IL-1β.

Conclusions: Distinct low doses of X-rays induce an anti-inflammatory phenotype of activated macrophages by lowering the amount of secreted IL-1β in a NF-κB dependent manner.     

Macrophage subtyping in the determination of age of injection sites

Determination of the age of injection marks in skin may be of particular interest in the investigation of drug abuse-related fatalities. The aim of our study was to assess the value of macrophage subtyping by antibodybased markers in the determination of the age of injection marks. Immunohistochemical investigations were performed with the antibodies Ki-MIP, 27E10, MRP14, MRP8 and 25F9. Monocytes/macrophages in acute lesions (several hours to 2 days old) expressed proteins detectable with the antibodies 27E10 and MRP14 and showed acute erythrophagia. An additional reaction with the antibody MRP8 was seen in lesions a few days old. An antigen recognized by the antibody 25F9 was found in tissue macrophages, multinucleated giant cells of active granulomas and siderophages. The expression of the 25F9 detectable antigen was absent in inactive granulomas and siderophages, whereas the macrophages were always detectable with the pan-macrophage marker Ki-M1P.     

A human ferritin iron oxide nano‐composite magnetic resonance contrast agent

Macrophages play important roles in the immunological defense system, but at the same time they are involved in inflammatory diseases such as atherosclerosis. Therefore, imaging macrophages is critical to assessing the status of these diseases. Toward this goal, a recombinant human H chain ferritin (rHFn)‐iron oxide nano composite has been investigated as an MRI contrast agent for labeling macrophages. Iron oxide nanoparticles in the form of magnetite (or maghemite) with narrow size distribution were synthesized in the interior cavity of rHFn. The composite material exhibited the R₂ relaxivity comparable to known iron oxide MRI contrast agents. Furthermore, the mineralized protein cages are readily taken up by macrophages in vitro and provide significant T2* signal loss of the labeled cells. These results encourage further investigation into the development of the rHFn‐iron oxide contrast agent to assess inflammatory disease status such as macrophage‐rich atherosclerotic plaques in vivo. Magn Reson Med 60:1073–1081, 2008. © 2008 Wiley‐Liss, Inc.     

Diagnostic significance of the histopathology of bone marrow macrophages in forensic autopsies

Forensic pathologists often encounter autopsies that require an assessment of antemortem general conditions (e.g., infection, metabolic disorders). To establish evaluation clues for such cases, we quantitatively examined macrophages and the general pathology of bone marrow in samples from 180 forensic autopsy cases of decedents with various conditions. Hematoxylin-eosin staining, Berlin blue staining, and immunostainings for CD163, CD138, and CD61 were performed. We determined the numbers per field (density) of total macrophages, swollen macrophages, macrophages with hemophagocytosis, and hemosiderin-laden macrophages. Each density was standardized by identifying its ratio to the total number of macrophages. The decedents' background data (cause of death, other pathological findings, postmortem interval, antemortem symptoms, and presence of resuscitation) were extracted. No correlations were found between the postmortem interval and the other decedent data, indicating that these data are not affected by postmortem changes. In the group in which inflammatory disease was the cause of death, there were significant elevations in the ratio of the swollen macrophage density to total macrophages. Significantly higher ratios of the density of swollen and hemophagocytic macrophages were observed in the group in which conditions with a prolonged agonal period were the cause of death. The group with a return of spontaneous circulation to resuscitation showed a significantly higher ratio of macrophage density with hemophagocytosis. This study provides the first statistical analysis focused on bone marrow histopathology in forensic autopsies. The results will be useful for elucidating causes of death and agonal-period conditions.     

He—Ne激光对小鼠腹腔巨噬细胞化学发光的影响

用6.1J/cm~2和4.3J/cm~2剂量的He-Ne激光分别辐照小鼠腹部,通过化学发光法测定腹腔巨噬细胞(PMΦ)依氧性吞噬杀菌功能,并作了动态观察。结果表明:激光照射组PMΦ的化学发光(CL)峰值、积分值和吞噬指数明显增高;与对照组相比,差异显著或非常显著,且两个照射组之间也有很显著的差异,第二组的免疫效应比第一组更显著。这提示,适当剂量的He-Ne激光辐照能促进PMΦ的呼吸爆发,增强依氧性吞噬杀菌作用,提高机体的细胞免疫功能。但考虑到He-Ne激光照射小鼠腹部时,小鼠腹壁很薄,激光可能直接达到腹腔内的组织;He-Ne激光对人的体表照射结果如何,尚待深入研究。     

烧伤小鼠腹腔巨噬细胞肌醇脂质信号系统变化的实验研究

目的：探讨烧伤导致巨噬细胞功能异常的发生机制。方法：测定了严重烧伤小鼠腹腔巨噬细胞（ＰＭΦ）磷酯酶Ｃ（ＰＬＣ）、甘油二酯（ＤＡＧ）、蛋白激酶Ｃ（ＰＫＣ）的生,三磷酸肌醇（ＩＰ３）、钙离子（Ｃａ＾２＋）、ＴＮＦ的变化,以及ＰＫＣ抑制剂Ｈ－７和钙调素（ＣａＭ）拮抗剂Ｗ－７对ＴＮＦ产生的影响。结果;上述所观察的指标在严重烧伤后６ｈｒ、１２ｈｒ、２４ｈｒ、都发生了非常明显的变化,ＰＫＣ抑制剂Ｈ－７能够显