Effects of cobalt and chromium ions on glycolytic flux and the stabilization of hypoxia‐inducible factor‐1α in macrophages in vitro

Implant wear and corrosion have been associated with adverse tissue reactions that can lead to implant failure. Wear and corrosion products are therefore of great clinical concern. For example, Co²⁺ and Cr³⁺ originating from CoCrMo‐based implants have been shown to induce a proinflammatory response in macrophages in vitro. Previous studies have also shown that the polarization of macrophages by some proinflammatory stimuli is associated with a hypoxia‐inducible factor‐1α (HIF‐1α)‐dependent metabolic shift from oxidative phosphorylation (OXPHOS) towards glycolysis. However, the potential of Co²⁺ and Cr³⁺ to induce this metabolic shift, which plays a determining role in the proinflammatory response of macrophages, remains largely unexplored. We recently demonstrated that Co²⁺, but not Cr³⁺, increased oxidative stress and decreased OXPHOS in RAW 264.7 murine macrophages. In the present study, we analyzed the effects of Co²⁺ and Cr³⁺ on glycolytic flux and HIF‐1α stabilization in the same experimental model. Cells were exposed to 6 to 24 ppm Co²⁺ or 50 to 250 ppm Cr³⁺. Glycolytic flux was determined by analyzing extracellular flux and lactate production, while HIF‐1α stabilization was analyzed by immunoblotting. Results showed that Co²⁺, and to a lesser extent Cr³⁺, increased glycolytic flux; however, only Co²⁺ acted through HIF‐1α stabilization. Overall, these results, together with our previous results showing that Co²⁺ increases oxidative stress and decreases OXPHOS, suggest that Co²⁺ (but not Cr³⁺) can induce a HIF‐1α‐dependent metabolic shift from OXPHOS towards glycolysis in macrophages. This metabolic shift may play an early and pivotal role in the inflammatory response induced by Co²⁺ in the periprosthetic environment.     

淫羊藿醇提物调节M2样巨噬细胞极化减少乳腺癌细胞迁移和集落形成的研究

目的 探讨淫羊藿Epimedium brevicornu醇提物对体外巨噬细胞极化调控机制及对M2样巨噬细胞促进4T1细胞迁移和集落形成的影响。方法 采用CCK-8法检测不同浓度淫羊藿醇提物对骨髓来源巨噬细胞（bonemarrowderived macrophages,BMDM）活力的影响,确定淫羊藿醇提物的安全作用浓度;采用Western blotting和q RT-PCR检测淫羊藿醇提物对M2样巨噬细胞标志物精氨酸酶-1(arginase-1,Arg-1)和CD163表达的影响;采用q RT-PCR检测淫羊藿醇提物对M2-肿瘤相关巨噬细胞（tumor-associated macrophages,TAMs）相关基因炎症区域1(found in inflammatory zone 1,Fizz1)、过氧化物酶体增殖物激活受体γ(peroxisome proliferator-activated receptor γ,PPARγ)、几丁质酶3样分子3(chitinase 3-like 3,Ym1)、趋化因子24(C-C motif chemokine ligand 24,CCL24)、巨...     

Inflammation,immunity and allergy

Injury or foreign invasion will instigate a cascade of events directed at eliminating the intruder and augmenting the healing process. This involves the unification of two separate processes (inflammatory and immune processes) to provide an effective host defence. Chemical mediators converge on the site of tissue damage and exert local and distant effects. The immune response is divided into innate and acquired immunity. The immediate, non-specific innate response, combined with the specifically targeted acquired response, provide our major defence mechanisms. Lymphocytes and immunoglobulins are the hallmark of acquired immunity. Regulation of these interlinked systems provide cohesion and a group of soluble proteins called cytokines have a major role. Protective immune mechanisms can sometimes cause detrimental effects to the host. We discuss and classify allergic reactions, in particular, the most severe and potentially life threatening form – anaphylaxis.     

LncRNA PCED1B-AS1调控NLRP3促进巨噬细胞清除结核分枝杆菌的机制研究

目的　研究长链非编码RNA PCED1B-AS1调控核苷酸结合寡聚化结构域样受体3（NLR family pyrindomain containing 3, NLRP3）对巨噬细胞清除结核分枝杆菌和分泌炎症因子的影响。方法　在巨噬细胞RAW264.7中转染pcDNA-PCED1B-AS1,并用结核分枝杆菌感染,qRT-PCR方法检测PCED1B-AS1和TNF-α、IL-6 mRNA水平,用菌落形成实验检测巨噬细胞对结核分枝杆菌的清除作用,Western blot方法检测细胞中NLRP3蛋白表达水平。将NLRP3 小干扰RNA（small interfering RNA, siRNA）和pcDNA-PCED1B-AS1共转染到巨噬细胞中,用结核分枝杆菌感染,同样检测细胞中TNF-α、IL-6 mRNA水平和菌落量。结果　结核分枝杆菌感染后的巨噬细胞中PCED1B-AS1水平降低,TNF-α、IL-6 mRNA水平升高。转染pcDNA-PCED1B-AS1后的巨噬细胞经过结核分枝杆菌感染以后,细胞中PCED1B-AS1、TNF-α、IL-6 mRNA水平升高,形成的菌落量减少,细胞中NLRP3蛋白表达水平升高。NLRP3 siRNA可以逆转过表达PCED1B-AS1对结核分枝杆菌感染的巨噬细胞中TNF-α、IL-6 mRNA表达和菌落量形成的影响。结论　上调PCED1B-AS1促进巨噬细胞清除结核分枝杆菌,诱导细胞表达TNF-α、IL-6 mRNA的机制与上调NLRP3表达有关。     

The use of P815: a warning

It is generally known that many malignancies are accompanied by forms of impairment of the immune system of the host. Also P815 mastocytoma cells can exert suppressive effects on the immune response reaction in vivo and in vitro. However, we have found a product released by P815 cells, which is able to activate macrophages non-specifically; macrophages normally not cytotoxic against tumor cells become cytotoxic against various tumor cell lines upon incubation with the P815 product. Nevertheless the same product which induces macrophage activation in vitro, seems to cause enhancement of tumor growth in vivo. We feel, if the activity of a tumor product depends so strongly on the experimental conditions used, one should be very careful in interpreting the results of experiments in which P815 cells are used as target cells.     

Caspase inhibitor Z-VAD-FMK potentiates heat shock-induced apoptosis and HSP70 synthesis in macrophages

Caspase inhibitor Z-VAD-FMK potentiated heat shock-induced apoptosis in macrophages. Z-VAD-FMK did not activate HSP70 synthesis, but significantly increased the intensity of this process during heat shock. It cannot be excluded that caspases abolish HSP70 accumulation under these conditions. The HSP70 synthesis inhibitor quercetin potentiated DNA fragmentation in macrophages cocultured with Z-VAD-FMK after heat shock. HSP70 play an important role in the protection of macrophages from caspase-independent apoptosis.Translated from Byulleten Eksperimentalnoi Biologii i Meditsiny, Vol. 138, No. 9, pp. 261–263, September, 2004     

Augmentation of recombinant CH50 polypeptide on the function of macrophages of mice during chemotherapy

CH50 is a Cell I- Hep Ⅱ bifunctional-domain recombinant polypeptide of human fibronectin expressed in E. colilll. This polypeptide can inhibit theinvasion and metastasis of tumor cells 1'n the[2] andactivate the anti--tumor activity of macrophages[']. Ithas been reported that this polypeptide andchemotherapeutic agent have a synergistic inhibitoryeffect on the metastasis of tumors[4J. In this study,we further investigated the effect of CH50 on thefunction of macrophages of mice during chem…     

不同临床表型COPD频繁和非频繁发作患者气道巨噬细胞的异质性

目的：比较频繁发作(frequent exacerbator,FE)和非频繁发作(infrequent exacerbator,iFE)的不同临床表型的 慢性阻塞性肺病(chronic obstructive pulmonary disease,COPD)患者的临床特征及诱导痰中巨噬细胞的异质性。方法： 分析80例慢性支气管炎(chronic bronchitis,CB)、肺气肿(emphysema,EM)、哮喘-COPD重叠(asthma-COPD overlap, ACO)表型的COPD急性发作住院患者的临床特征;采用流式细胞术检测诱导痰巨噬细胞CCL3和CD163的表达,定 量聚合酶链反应(quantitative PCR,qPCR)检测HIF-1α和Cav-1的表达。结果：FE和iFE患者在年龄,第1 s用力呼气容积 (forced expiratory volume in one second,FEV1)/用力肺活量(forced vital capacity,FVC),痰细菌阳性率,COPD评估测试 (COPD Assessment Test,CAT)评分,改良医学研究委员会(Modifi ed Medical Research Council,mMRC)评分方面的差异 有统计学意义(P<0.05);相对于iFE患者,FE患者诱导痰巨噬细胞上CCL3荧光强度明显降低(P<0.01),而CD163显著增 高(P<0.01),同时HIF-1α和Cav-1 mRNA水平也显著升高(P<0.01)。CB表型的FE患者与iFE患者之间在年龄,痰细菌阳 性率,CAT评分,mMRC评分方面的差异有统计学意义(P<0.05);相对于iFE患者,FE患者诱导痰巨噬细胞上CCL3荧 光强度轻度降低(P>0.05),而CD163显著增高(P<0.01),同时HIF-1α和Cav-1 mRNA水平也显著升高(P<0.01)。EM表型 的FE患者与iFE患者之间在年龄,病程,FEV1/FVC,痰细菌阳性率,CAT评分,mMRC评分方面的差异有统计学意义 (P<0.05);相对于iFE患者,FE患者诱导痰巨噬细胞CCL3荧光强度轻度减低(P>0.05),而CD163轻度升高(P>0.05),同 时HIF-1α水平轻度升高(P>0.05),Cav-1水平则显著增加(P<0.01)。ACO表型的FE与iFE患者所有临床特征差异均无统计 学意义,FE患者诱导痰巨噬细胞CCL3荧光强度明显低于iFE患者(P<0.01),而CD163差异无统计学意义(P>0.05),同 时HIF-1α(P<0.01)和Cav-1(P<0.05)表达也显著增加。全部FE及CB表型FE患者CCL3与HIF-1α和Cav-1均呈显著负相关, CD163仅与HIF-1α呈显著正相关;EM表型FE患者CD163与HIF-1α呈显著正相关;ACO表型FE患者CCL3与HIF-1α呈显 著负相关,而CD163与HIF-1α呈显著正相关。结论：CB,EM,ACO表型FE和iFE患者临床特征差异不一,诱导痰中替 代活化巨噬细胞(M2)在FE患者中占优势,HIF-1α可能在其极化过程中起关键作用。     

硝酸甘油、地塞米松对哮喘肺泡巨噬细胞源性一氧化氮、内皮素的影响

研究哮喘患者肺泡巨噬细胞 (AM)源性一氧化氮 (NO)、内皮素 (ET)的变化及硝酸甘油 (NTG)、地塞米松(DXM)对两者的影响及机制。对 15例轻、中度过敏性支气管哮喘发作期患者的AM(分为未干预组、DXM干预组、NTG干预组 ) ,7名健康自愿受试者的AM(未干预组 )培养 48h ,用镀铜镉还原法、放射免疫法和原位杂交法分别测定AM培养上清液中NO ,ET水平和iNOS mRNA ,ET mRNA的表达。结果发现 ,哮喘AMiNOS mRNA ,ET mRNA表达增强 ,分别导致NO ,ET水平升高 ;NTG以直接作用的方式促进AM源性NO的产生 ,反馈抑制iNOS mRNA表达并明显抑制ETmRNA的表达 ,降低ET的水平 ;DXM降低哮喘AM源性NO ,ET水平及iNOS mRNA ,ET mRNA的表达 ,尤以抑制iNOS mRNA表达和降低NO水平为甚 ,使NO ,ET处于低水平的异常状态。     

胆必清对内毒素诱导的大鼠腹腔巨噬细胞内游离钙浓度的影响

目的：探讨中药胆必清对内毒素诱导的大鼠腹腔巨噬细胞游离钙浓度的影响。方法：制备大鼠腹腔巨噬细胞并在体外进行孵育,使用荧光探针用荧光法测定钙浓度。巨噬细胞随机分为正常对照组,内毒素LPS组,小、中和大剂量中药组。结果：LPS能诱导巨噬细胞[Ca2^ ]i的升高。加入胆必清后,[Ca2^ ]i可有不同程度的降低。结论：胆必清能抑制LPS诱导的大鼠腹腔巨噬细胞内的[Ca2^ ]i升高,提示该药具有抗炎和免疫调节作用。