黄芪多糖联合布洛芬对创伤感染患者巨噬细胞分泌功能、T淋巴细胞亚群水平的影响及其影响因素分析

目的:探讨黄芪多糖联合布洛芬对创伤感染患者巨噬细胞分泌功能、T淋巴细胞亚群水平的影响及影响因素。方法:2014年3月-2018年3月我院收治创伤感染患者85例,其中重度感染39例,将所有患者按感染轻重进行平衡随机分为对照组43例和研究组42例,对照组应用布洛芬治疗,研究组应用黄芪多糖联合布洛芬治疗。比较两组患者巨噬细胞分泌功能、T淋巴细胞亚群水平,分析患者发生重度感染的影响因素及巨噬细胞分泌功能、T淋巴细胞亚群水平与创伤感染的关系。结果:治疗后研究组患者血清中肿瘤坏死因子(TNF)、白介素细胞-6(IL-6)和白介素细胞-1(IL-1)的水平低于对照组(P<0.05)。治疗后研究组外周血中CD3⁺、CD4⁺、CD8⁺和CD4⁺/CD8⁺水平高于对照组(P<0.05)。巨噬细胞分泌功能较高、T淋巴细胞亚群水平较低的患者发生重度感染的概率较高(P<0.05);巨噬细胞分泌功能与创伤感染呈正向关联关系(P<0.05,OR>1);T淋巴细胞亚群水平与创伤感染呈负向关联关系(P<0.05,OR<1);治疗方法以研究组方法为优(P<0.05,OR>1)。结论:黄芪多糖联合布洛芬治疗创伤感染患者效果更显著,巨噬细胞分泌功能、T淋巴细胞亚群水平均是创伤感染的影响因素。     

植物乳杆菌P-8对小鼠免疫功能的调节作用研究

摘要 目的：不同类型的益生菌株免疫调节功能各异。本文旨在评价植物乳杆菌P-8（Lactobacillus plantarum P-8）对小鼠免疫功能的调控作用及机制。方法：C57BL/6J小鼠每日灌胃给予不同剂量的植物乳杆菌P-8（0. 25 mg/kg、0.5 mg/kg、1.5 mg/kg）,连续30天,记录小鼠一般情况。给药结束后处死动物,测定小鼠脏器/体重比;小鼠碳廓清实验、小鼠腹腔巨噬细胞吞噬鸡红细胞实验评价各组小鼠的单核-巨噬细胞功能;血清溶血素测定、抗体生成细胞实验评价各组小鼠的体液免疫功能;脾淋巴细胞转化实验、迟发型变态反应实验评价各组小鼠的细胞免疫功能;NK细胞的活性测定实验评价小鼠的NK细胞活性。结果：与对照组相比,低、中、高剂量组植物乳杆菌P-8对小鼠脏器/体重比值差异无统计学意义（P>0.05）;且植物乳杆菌P-8可显著提高小鼠的碳廓清能力、小鼠腹腔巨噬细胞吞噬鸡红细胞能力、半数溶血值、二硝基氟苯诱导的小鼠迟发型变态反应及NK细胞活力（P均<0.05）。结论：植物乳杆菌P-8可通过提高单核-巨噬细胞功能、体液免疫功能、细胞免疫功能及NK细胞活力增强小鼠的免疫功能。     

Effective-components combination improves airway remodeling in COPD rats by suppressing M2 macrophage polarization via the inhibition of mTORC2 activity

BackgroundIn chronic obstructive pulmonary disease (COPD), M2 macrophages release multiple tissue repair-related factors, leading to airway remodeling, a significant pathological characteristic. Meanwhile, effective-components combination (ECC), derived from Bufei Yishen formula (BYF), is an effective treatment for COPD.PurposeTo determine the potential mechanisms of ECC in airway remodeling in COPD by suppressing M2 macrophage polarization.MethodsWe established a rat COPD Model using exposure to cigarette smoke and bacterial infection to investigate the efficacy of ECC. We also treated macrophages with IL-4 for 12 h to explore the in vivo effect of ECC on M2 macrophage polarization and mTORC2 signals.ResultsThe disease severity of COPD rats could be alleviated by ECC treatment, which improved pulmonary function and alleviated pathological injuries in lung tissue and the inflammatory cytokine levels. Meanwhile, ECC could ameliorate airway remodeling by reducing collagen deposition, hindering airway mucus hypersecretion and smooth muscle cell proliferation, and reducing the number of M2 macrophages in the lung tissues of COPD rats. Furthermore, with IL-4-induced macrophages, we found that ECC could suppress M2 macrophage polarization by decreasing the levels of M2 macrophage markers. Finally, we discovered that ECC inhibited mTORC2 activity by examining p-mTOR²⁴⁸¹ and its downstream protein p-Akt⁴⁷³.ConclusionsECC exerts beneficial effects on airway remodeling in COPD rats, likely by suppressing M2 macrophage polarization via the inhibition of mTORC2 activity.     

Pentraxin-3 regulates the inflammatory activity of macrophages

Background and aimsPentraxin-3 (PTX3) reportedly has protective roles in atherosclerosis and myocardial infarction, and is a useful biomarker of vascular inflammation. However, the detailed functions of PTX3 in inflammation are yet to be elucidated. This study aimed to investigate the function of PTX3 in macrophages.MethodsPMA-treated THP-1 cell line (THP-1 macrophage) and monocyte-derived human primary macrophages were treated with recombinant PTX3. Cytokine and chemokine levels in the THP-1 culture medium were measured as well as monocyte chemoattractant protein (MCP-1) concentrations in the Raw 264.7 cell culture medium. PTX3-silenced apoptotic macrophages (THP-1 cell line) were generated to investigate the roles of PTX3 in phagocytosis.ResultsIn the presence of PTX3, macrophage interleukin-1β (IL-1β), tumor necrosis factor-alpha (TNF-α) and MCP-1 levels were reduced significantly (?39%, P=0.007; ?21%, P=0.008; and ?67%, P=0.0003, respectively), whilst activated transforming growth factor-β (TGF?β) was detected in the THP-1 macrophages (P=0.0004). Additionally, PTX3 induced Akt phosphorylation and reduced nuclear factor-kappa B (NF-κB) activation by 35% (P=0.002), which was induced by TNF-α in THP-1 macrophages. Furthermore, silencing of PTX3 in apoptotic cells resulted in increased macrophage binding, elevated expression rate of HLA-DR (+30%, P=0.015) and CD86 (+204%, P=0.004) positive cells, and induction of IL-1β (+36%, P=0.024) production. Conversely, adding recombinant PTX3 to macrophages reduced CD86 and HLA-DR expression in a dose-dependent manner.ConclusionsWe identified PTX3 as a novel regulator of macrophage activity, and this function suggests that PTX3 acts to resolve inflammation.     

巨噬细胞来源外泌体介导心肌梗塞后心脏重塑的作用研究

目的:研究活化的巨噬细胞来源外泌体在心肌梗塞后心脏重塑中的作用。方法:采用超高速离心分离提取溶血磷脂酸作用下巨噬细胞来源的外泌体,将其与心脏成纤维细胞共同孵育48小时,利用Edu细胞增殖实验、Transwell实验及免疫荧光等方法检测溶血磷脂酸刺激(LPS)下巨噬细胞来源外泌体对心脏成纤维细胞的增殖、迁移以及分化的影响。选取正常C57雄性小鼠32只,根据其是否结扎左侧冠状动脉前降支及是否进行心脏原位外泌体注射,将实验小鼠随机分为:正常组,假手术组,心梗+空白外泌体组及心梗组+LPS刺激外泌体组。手术完成4周后行心脏超声、Masson染色以检测各组实验小鼠心功能状态及心脏纤维化程度。结果:在细胞实验中,LPS刺激的巨噬细胞来源外泌体可以显著增加心脏成纤维细胞的增殖、迁移以及分化能力;在动物实验中,相对于正常组、假手术组及心梗+空白-外泌体组,心梗+LPS-外泌体组小鼠的左心室射血分数及短轴收缩率显著下降,左心室舒张末及收缩末内径显著增加。Masson染色检测提示心肌梗塞+LPS-外泌体组小鼠心脏纤维化程度显著高于其余三组。结论:活化的巨噬细胞来源的外泌体可以显著加速心梗后心脏重塑的进程。     

巨噬细胞膜仿生纳米铁颗粒制备及多形性胶质母细胞瘤MRI成像的初步实验研究

摘要 目的：探讨巨噬细胞膜仿生的纳米铁颗粒（Fe3O4 NCs@MM）对多形性胶质母细胞瘤MRI成像的研究。方法：制备巨噬细胞膜仿生的纳米铁颗粒Fe₃O₄ NCs@MM,利用动态光散射（Dynamic Light Scattering,DLS）和透射电子显微镜（Transmission Electron Microscope,TEM）对其水合动力学粒径、表面电势和形态进行表征。采用SDS-聚丙烯酰胺凝胶电泳（sodium dodecyl sulphate-polyacrylamide gel electrophoresis,SDS-PAGE）评价巨噬细胞膜的完整包覆;紫外可见光谱测定巨噬细胞膜仿生的纳米铁颗粒抗蛋白吸附能力。通过MRI成像系统,分析了含不同浓度的Fe元素（0.1-1.6 mM）的Fe₃O₄ NCs@MM在GSH存在或不存在时的T₁弛豫效应。采用细胞增殖-毒性实验（Cell Counting Kit-8,CCK-8）,测定巨噬细胞膜仿生纳米铁颗粒处理肿瘤细胞24 h后的细胞活性。尾静脉注射巨噬细胞膜仿生纳米铁颗粒至原位胶质母细胞瘤模型中,观察成像效果。结果：巨噬细胞膜仿生的纳米铁颗粒Fe₃O₄ NCs@MM的水合动力学粒径和表面电势分别为 286.5±7.6 nm和-20.7±3.5 mV,且在水溶液中分布均匀,具有较好的单分散性。包覆巨噬细胞膜的纳米铁颗粒具备抗蛋白吸附的能力。MRI成像显示,制备的巨噬细胞膜仿生的纳米铁颗粒Fe₃O₄ NCs@MM为GSH响应型MRI对比剂,具有较好的T₁-加权磁共振成像效果,在尾静脉注射巨噬细胞膜的纳米铁颗粒0.5 h后,肿瘤部位的信号可见增强。结论：巨噬细胞膜仿生的纳米铁颗粒Fe₃O₄ NCs@MM可实现多形性胶质母细胞瘤的MRI成像。     

Macrophage secretions modulate the steroidogenesis of polycystic ovary in rats: effect of testosterone on macrophage pro-inflammatory cytokines

AimsThe macrophage secretions' effect on ovarian steroidogenesis is investigated in a polycystic ovary syndrome rat model (PCO rat). The influence of testosterone environment on the expression of macrophage pro-inflammatory cytokines that participate in ovarian steroidogenesis is studied.Main methodsPCO rats were induced by estradiol valerate. Spleen macrophages were cultured with and without testosterone (10^? 6 M) and their secretions were used to stimulate ovaries from PCO and control rats. Ovarian hormones released and ovary mRNA levels of P450 aromatase and 3β-hydroxysteroid dehydrogenase were measured by radioimmunoassay and RT-PCR, respectively. The tumor necrosis factor alpha (TNFα) and nitric oxide (NO) levels in macrophage culture medium, along with the TNFα, interleukin (IL)-6, IL-10 and androgen receptors (AR) mRNA levels in macrophage cells were determined.Key findingsMacrophages from PCO rats released more TNFα and NO, expressed higher TNFα and IL-6, lower AR, and no change in IL-10 mRNA levels than control macrophages. TNFα, IL-6 and AR changes were greater after macrophage testosterone treatment. Macrophage secretions from PCO rats stimulated androstenedione and decreased estradiol release and ovarian mRNA P450 aromatase expression in PCO rats compared to macrophage secretions from control rats.These effects were greater when macrophages from PCO rats were treated with testosterone. Ovarian progesterone response was unchanged.SignificanceThe differential steroidogenic ability of macrophage secretions from PCO rats is associated to the in vitro testosterone environment. Testosterone, probably acting on macrophage AR, induces a greater release of TNFα, modifying ovarian response by increasing androstenedione and slightly decreasing estradiol without affecting progesterone.     

双氢青蒿素调控巨噬细胞增殖和迁移的作用研究

目的:探讨双氢青蒿素在体外对小鼠单核巨噬细胞RAW264.7的增殖、克隆形成、周期、凋亡和迁移的影响。方法:采用梯度浓度(2.5μg/m L, 5μg/m L, 10μg/m L, 20μg/m L)的双氢青蒿素处理RAW264.7细胞,利用CCK8实验检测双氢青蒿素对巨噬细胞增殖能力的影响,利用克隆形成实验检测双氢青蒿素对RAW264.7细胞克隆形成能力的影响,利用流式细胞术检测双氢青蒿素对RAW264.7细胞周期和凋亡的影响,利用划痕修复实验检测RAW264.7细胞迁移能力。结果:CCK8实验结果显示,双氢青蒿素可以显著抑制RAW264.7巨噬细胞的增殖能力,且抑制效果与双氢青蒿素的浓度呈正相关性。克隆形成实验结果显示,双氢青蒿素可以抑制细胞的克隆形成能力。双氢青蒿素处理使RAW264.7细胞G0/G1期比例显著升高,S期与G2/M期细胞比例显著降低。双氢青蒿素对巨噬细胞凋亡具有诱导作用,且凋亡诱导作用呈现浓度依赖的特性。划痕修复实验结果显示,双氢青蒿素可以显著抑制RAW264.7巨噬细胞的迁移能力。结论:双氢青蒿素可以导致巨噬细胞的细胞周期G0/G1阻滞,并且诱导细胞凋亡,对巨噬细胞增殖和迁移具有抑制作用。     

新型PEI衍生物的体外表征及对巨噬细胞的转染效果研究

目的:以PEI1.8kDa为基础连接戊二醛合成新型可降解PEI衍生物GPEI后,研究GPEI包裹质粒转染巨噬细胞后对巨噬细胞合成蛋白及表型的影响。方法:合成GPEI后,检测了GPEI质粒聚合物的粒径及电位,并通过透射电镜观察聚合物形态,通过免疫荧光、WB、RT-PCR、Elisa及Transwell实验检测巨噬细胞RNA、蛋白、终产物水平变化及表型改变。结果:GPEI能够包裹质粒,形成100 nm左右带有正电荷的聚合物,WB、RT-PCR结果表明GPEI转染较PEI25kDa有优势(P0.05),巨噬细胞转染后6天持续分泌PGI2(P0.05)。结论:以PEI1.8kDa为基础合成的GPEI能够包裹PTGIS质粒并将其转染进巨噬细胞后对巨噬细胞的表型有明显影响。     

Mesenchymal stromal cells regulate THP-1–differentiated macrophage cytokine production by activating Akt/mammalian target of rapamycin complex 1 pathway

Background aimsThe Akt/mammalian target of rapamycin (mTOR) pathway in macrophages converges inflammatory and metabolic signals from multiple receptors to regulate a cell's survival, metabolism and activation. Although mesenchymal stromal cells (MSCs) are well known to modulate macrophage activation, the effects of MSCs on the Akt/mTOR pathway in macrophages have not been elucidated.MethodsWe herein investigated whether MSCs affect the Akt/mTOR complex 1 (mTORC1) pathway to regulate macrophage polarization.ResultsResults showed that human bone marrow–derived MSCs induced activation of Akt and its downstream mTORC1 signaling in THP-1–differentiated macrophages in a p62/sequestosome 1–independent manner. Inhibition of Akt or mTORC1 attenuated the effects of MSCs on the suppression of tumor necrosis factor-α and interleukin-12 production and the promotion of interleukin-10 and tumor growth factor-β1 in macrophages stimulated by lipopolysaccharide/ATP. Conversely, activation of Akt or mTORC1 reproduced and potentiated MSC effects on macrophage cytokine production. MSCs with cyclooxygenase-2 knockdown, however, failed to activate the Akt/mTORC1 signaling in macrophages and were less effective in the modulation of macrophage cytokine production than control MSCs.ConclusionsThese data demonstrate that MSCs control THP-1–differentiated macrophage activation at least partly through upregulation of the Akt/mTORC1 signaling in a cyclooxygenase-2–dependent manner.     

Lack of a significant role of P-Rex1, a major regulator of macrophage Rac1 activation and chemotaxis,in atherogenesis

BackgroundRho GTPases are known to play important roles in regulating multiple cellular processes that include cell polarization and migration. Among these Rho GTPases, Rac has been shown to be essential for F actin formation and cell migration. P-Rex1 is a guanine nucleotide exchange factor (GEF) that was previously found to mediate the activation of Rac2, but not Rac1, in mouse neutrophils.ObjectivesHere we examined the role of P-Rex1 in mouse macrophages and atherogenesis.Methods and resultsPBD (p21 binding domain) pull down assay was performed to compare the Rac1 activation in WT and P-Rex1-deficient macrophage. In addition, transwell assay was conducted to compare chemotaxis of WT and P-Rex1-deficient macrophage. We found that P-Rex1 is a major Rac1 regulator in mouse macrophages as its deficiency significantly compromises macrophage chemotaxis, superoxide production (SOD), and Rac1 activation in response to chemoattractants. The potential role of P-Rex1 in atherogenesis is also investigated by transferring P-Rex1-deficient bone marrow cells to LDLR deficient mice. Contrary to our prediction, P-Rex1 deficiency did not alter atherogenesis, suggesting chemoattractant-induced macrophage migration may not have a significant role in atherogenesis.ConclusionsP-Rex1 is one of the major GEFs in macrophage regulating Rac1 activation and chemotaxis.     

不同盐浓度对盐敏感性高血压肾脏纤维化及巨噬细胞浸润的影响

目的:探讨不同盐浓度对盐敏感高血压肾脏纤维化及及巨噬细胞浸润的影响。方法:将6周龄Dahl盐敏感大鼠随机分为正盐组(0.3%氯化钠)、4%氯化钠组、8%氯化钠组三组,每组15只,持续喂养8周。在8周时间点应用tail-cuff method方法监测鼠尾血压,Masson三原色法检测三组大鼠肾脏组织纤维化情况,免疫组化及Western blotting方法检测各组大鼠肾脏巨噬细胞浸润情况。结果:1)4%高盐组及8%高盐组大鼠8周时间点血压均较正盐组现在升高,8%高盐组大鼠血压较4%高盐组进一步升高。2)4%高盐组及8%高盐组大鼠8周时间点肾脏相对重量及肾脏纤维化程度较正盐组增加,8%高盐组较4%高盐组进一步增加。3)4%高盐组及8%高盐组大鼠8周时间点肾脏组织巨噬细胞浸润增加,8%高盐组较4%高盐组进一步增加。结论:盐浓度不同对盐敏感性肾脏纤维化及巨噬细胞浸润程度影响不同,盐浓度升高可加重盐敏感高血压时肾脏纤维化及及巨噬细胞浸润。     

Serum from patients with asthma potentiates macrophage phagocytosis and human mesenchymal stromal cell therapy in experimental allergic asthma

Background/AimsAlthough several studies have demonstrated that mesenchymal stromal cells (MSCs) exhibit beneficial immunomodulatory properties in preclinical models of allergic asthma, effects on airway remodeling have been controversial. Recent evidence has shown that MSCs modify their in vivo immunomodulatory actions depending on the specific inflammatory environment encountered. Accordingly, we assessed whether the therapeutic properties of human mesenchymal stromal cells (hMSCs) could be potentiated by conditioning these cells with serum (hMSC-serum) obtained from patients with asthma and then transplanted in an experimental model of house dust mite (HDM)-induced allergic asthma.MethodshMSC and hMSC-serum were administered intratracheally 24 h after the final HDM challenge. hMSC viability and inflammatory mediator production, lung mechanics and histology, bronchoalveolar lavage fluid (BALF) cellularity and biomarker levels, mitochondrial structure and function as well as macrophage polarization and phagocytic capacity were assessed.ResultsSerum preconditioning led to: (i) increased hMSC apoptosis and expression of transforming growth factor-β, interleukin (IL)-10, tumor necrosis factor-α–stimulated gene 6 protein and indoleamine 2,3-dioxygenase-1; (ii) fission and reduction of the intrinsic respiratory capacity of mitochondria; and (iii) polarization of macrophages to M2 phenotype, which may be associated with a greater percentage of hMSCs phagocytosed by macrophages. Compared with mice receiving hMSCs, administration of hMSC-serum led to further reduction of collagen fiber content, eotaxin levels, total and differential cellularity and increased IL-10 levels in BALF, improving lung mechanics. hMSC-serum promoted greater M2 macrophage polarization as well as macrophage phagocytosis, mainly of apoptotic hMSCs.ConclusionsSerum from patients with asthma led to a greater percentage of hMSCs phagocytosed by macrophages and triggered immunomodulatory responses, resulting in further reductions in both inflammation and remodeling compared with non-preconditioned hMSCs.     

Mejoría de la inmunidad y la esperanza de vida en ratones maduros,genéticamente deficientes en la síntesis de catecolaminas,tras convivir con controles durante dos meses

IntroductionMice hemizygous in tyrosine hydroxylase (TH-HZ), the limiting enzyme in catecholamine synthesis, show premature immunosenescence, which in females is associated with a shorter lifespan than the corresponding controls (WT). The coexistence of TH-Hz with WT improves the immune function in both males and females in adulthood.ObjectiveTo test whether cohabitation for two months of mature male TH-HZ with WT improves the immune function of the former and whether this impacts the lifespan.Material and methodsMature male ICR-CD1 mice (13 ± 1 months) TH-HZ coexisted with WT (2:4 ratio in each cage) for two months. Peritoneal leukocytes were extracted from all animals at baseline, one month, and two months after cohabitation, and macrophage phagocytic capacity, macrophage and lymphocyte chemotaxis, natural killer (NK) antitumor activity, and lymphoproliferative capacity in response to the mitogens concanavalin A and lipopolysaccharide (LPS) were assessed. The animals were maintained under these conditions until their natural death.ResultsThe TH-HZ, which start, in general, with lower values than the WT in the immune functions studied, improved them after two months of cohabitation, becoming similar to those of the controls. This improvement was already observed in NK activity after one month of cohabitation.The TH-HZ presented lower mean longevity than WT, but when they cohabited with WT, it was similar to the latter.ConclusionThe coexistence of TH-HZ male mice with WT mice for two months at mature age improves these genetically modified animals’ immune response and longevity.     

巨噬细胞膜伪装纳米颗粒的制备和功能表征

摘要 目的：巨噬细胞具有炎症趋化能力,近年来巨噬细胞膜伪装的纳米递送载体引起研究者的广泛关注。本文提供了一种巨噬细胞膜伪装纳米颗粒的方法,即摄取-挤出法,并对该法制得的纳米颗粒进行表征,考察纳米颗粒在不同细胞中的摄取。方法：利用溶胶-凝胶法制备装载阿霉素的介孔硅（DMSN）纳米颗粒,再利用RAW 264.7巨噬细胞吞噬DMSN,最后将巨噬细胞连续挤出制得巨噬细胞膜伪装的载有阿霉素的介孔硅（DMSN@CM）纳米颗粒。动态光散射激光粒度仪（DLS）测定DMSN@CM颗粒的粒径和表面电位,透射电子显微镜（TEM）观察纳米颗粒形态,聚丙烯酰胺凝胶电泳（SDS-PAGE）验证细胞膜的成功伪装。然后通过激光共聚焦显微镜与流式细胞术共同考察了DMSN@CM在不同细胞中的摄取情况。结果：成功制备了DMSN和DMSN@CM纳米颗粒。DMSN粒径为116.7±3.2 nm,zeta表面电势为 -29.5± 1.3 mV;MSN@CM粒径为128.0±9.3 nm,zeta表面电势为 -26.7 ±1.2 mV。TEM与SDS-PAGE共同验证了DMSN@CM表面细胞膜的成功包覆。细胞摄取试验表明巨噬细胞膜的伪装可以抑制RAW 264.7细胞对DMSN@CM的摄取;促进MDA-MB-231细胞对DMSN@CM的摄取。结论：利用摄取-挤出法成功构建了DMSN@CM纳米颗粒,该法简便高效,为纳米颗粒的细胞膜伪装提供了一种新的手段。     

乙酰辅酶A羧化酶抑制剂联合非诺贝特对小鼠非酒精性脂肪肝的治疗效果

摘要 目的：探讨乙酰辅酶A羧化酶抑制剂（MK-4074）联合非诺贝特对小鼠非酒精性脂肪肝（NAFLD）的脂质含量以及肝功能的改善效果。方法：20只C57BL/6小鼠给予60%高脂饲料连续喂养8周构建NAFLD小鼠模型后，随机分为安慰剂组、MK-4074组、非诺贝特组以及MK-4074联合非诺贝特治疗组，每组各5只，继续高脂喂养并分别给予安慰剂（Placebo）、MK-4074（10 mg/kg/天）、非诺贝特（30 mg/kg/天）、以及MK-4074（10 mg/kg/天）+ 非诺贝特（30 mg/kg/天）治疗持续8周。治疗结束后对小鼠体重、肝指数、肝脏脂质含量、肝功能以及肝脏病理和肝脏中性粒细胞和巨噬细胞浸润情况进行分析。结果：与安慰剂组相比，单用MK-4074治疗可显著降低肝指数、肝脏甘油三酯（TG）、胆固醇（TC）、非酯化脂肪酸（NEFA）的含量以及血清ALT和AST水平，而对小鼠体重和血清TC没有显著影响；单用非诺贝特可显著降低小鼠体重，肝脏TG、TC、NEFA以及血清TG、 ALT和AST水平，对小鼠的肝指数、血清TC没有显著影响；而MK-4074与非诺贝特联合治疗可显著降低小鼠体重、肝脏TG、TC、NEFA，以及血清TG、ALT和AST水平，降低肝脏脂质积累以及中性粒细胞与巨噬细胞浸润，效果优于MK-4074或非诺贝特单药治疗。结论：MK-4074联合非诺贝特可显著减少NAFLD小鼠肝脏的脂质含量，改善肝功能。     

β-caryophyllene ameliorated obesity-associated airway hyperresponsiveness through some non-conventional targets

BackgroundObesity worsens airway hyperresponsiveness (AHR) in asthmatic subjects by up-regulating macrophage polarization that leads to excessive secretion of pro-inflammatory adipokines from white adipose tissue followed by generation of oxidative stress in the respiratory system. Treatment through conventional signaling pathways proved to be inadequate in obese asthmatics, so a therapeutical approach through a non-conventional pathway may prove to be effective.PurposeThis study aimed to investigate the efficacy of a FDA-approved food additive, β-caryophyllene (BCP) in obesity-associated AHR.MethodA repertoire of protein expression, cytokine and adiponectin estimation, oxidative stress assays, histopathology, and fluorescence immune-histochemistry were performed to assess the efficacy of BCP in C57BL/6 mice model of obesity-associated AHR. Additionally, human adipocyte was utilized to study the effect of BCP on macrophage polarization in Boyden chamber cell culture inserts.ResultsObesity-associated AHR is ameliorated by administration of BCP by inhibition of the macrophage polarization by activation of AMPKα, Nrf2/HO-1 and AdipoR1 and AdipoR2 signaling pathway, up-regulation of adiponectin, GLP-1, IFN-γ, SOD, catalase and down-regulation of NF-κB, leptin, IL-4, TNF, and IL-1β. Browning of eWAT by induction of thermogenesis and activation of melanocortin pathway also contributed to the amelioration of obesity-associated AHR. We conclude that BCP ameliorated the obesity-associated AHR via inhibition of macrophage polarization, activation of AMPKα, Nrf2/HO-1, and up-regulation of AdipoR1 and AdipoR2 expression and down-regulation of NFκB expression in lung of animal.ConclusionBeing an FDA-approved food additive, BCP may prove to be a safe and potential agent against obesity-associated AHR.     

miR-448-3p通过调节KLF5的表达抑制颅内动脉瘤进展

目的:明确mi R-448-3p对颅内动脉瘤发展的影响。方法:我们通过结扎左侧肾动脉和左侧颈总动脉的方法建立大鼠IA模型;qRT-PCR用于检测mi R-448-3p表达;qRT-PCR和western blot用于检测KLF5 m RNA和蛋白表达;qRT-PCR和ELISA法用于检测炎症因子水平。结果:我们发现IA诱导大鼠中mi R-448-3p表达下调,而KLF5表达上调。我们发现并鉴定出KLF5是平滑肌细胞中mi R-448-3p的直接靶点。此外,mi R-448-3p处理使得IA诱导4周后动脉瘤大小和瘤腔截面积变小。mi R-448-3p处理保护了IA诱导后的壁厚比,抑制了巨噬细胞浸润。IAs引起KLF5表达显著增加,被mi R-448-3p处理后表达显著降低。我们还发现mi R-448-3p在脂多糖诱导的RAW 264.7巨噬细胞中具有抗炎作用。脂多糖促进KLF5、MMP2、MMP9的表达水平,但却被mi R-448-3p所抑制。结论:研究结果表明,mi R-448-3p可以抑制IA的进展,其机制可能是通过下调KLF5的介导的炎症反应。     

Effect of Fonsecaea monophora on the Polarization of THP-1 Cells to Macrophages

Background

Chromoblastomycosis is a chronic, progressive fungal disease of the skin and subcutaneous tissue caused by a unique group of dematiaceous fungi. Fonsecaea monophora, a new species distinct from Fonsecaea pedrosoi strains, is the main pathogen responsible for chromoblastomycosis in south China. Macrophages can be polarized into two categories: classically activated and alternatively activated.

Objectives

Little is known about the relationship between F. monophora and macrophage polarization. This study aimed to study the effect of F. monophora on the polarization of THP-1 cells to macrophages.

Methods

We established coculture systems of F. monophora and THP-1-derived macrophages in different activation states.

Results

F. monophora enhanced the phagocytosis by macrophages in the initially activated state and weakened the phagocytosis by classically activated macrophages without affecting that by alternatively activated macrophages. Classically activated macrophages had the strongest killing effect on F. monophora, while the initially activated macrophages had the weakest. The pathogen could not be rapidly cleared by any type of macrophage. F. monophora promoted the expression of proinflammatory cytokines and inhibited that of anti-inflammatory cytokines.

Conclusions

F. monophora promoted the polarization of THP-1 cells to classically activated macrophages and inhibited that of THP-1 cells to alternatively activated macrophages.