野漆树苷对LPS诱导的RAW264.7细胞的抗炎作用及机制探究

目的:探究野漆树苷(Rhoifolin,RHO)对细菌脂多糖(lipopolysaccharide,LPS)诱导的小鼠单核巨噬细胞RAW264.7细胞炎症模型释放炎性因子的影响,并探讨其作用机制.方法:用脂多糖(0.5μg·mL~(-1))刺激体外生长良好的RAW264.7细胞24h建立体外细胞炎症模型,以MTT法测定不同浓度RHO对RAW264.7细胞的毒性作用,用Griess试剂法检测一氧化氮(Nitric Oxide NO)的含量,RT-PCR法检测细胞中肿瘤坏死因子α(Tumor Necrosis Factor,TNF-α),白介素1β(Interleukin-1β,IL~(-1)β),白介素6(Interleukin,IL-6)的含量,采用Western Blot法检测MAPK信号通路中相关蛋白的含量.结果:与空白对照组相比,在LPS诱导下,RAW264.7细胞分泌致炎因子NO,TNF-α,IL~(-1)β,IL-6(P0.01);与模型组相比,25~100μmol·L~(-1)的RHO可明显下调LPS诱导的RAW264.7细胞释放炎症因子NO,TNF-α,IL~(-1)β和IL-6(P0.05,P0.01),并呈现良好的剂量依赖关系;野漆树苷还不同程度的抑制了Erk和JNK激酶的磷酸化.结论:RHO可以抑制LPS所致的RAW264.7细胞炎症反应,减少炎症因子NO分泌,抑制TNF-α,IL~(-1)β和IL-6mRNA的合成,抑制JNK/SAPK及Erk信号通路可能是其抗炎作用机制之一.     

脂多糖对RAW264.7细胞IL-10分泌及mRNA表达的影响

目的探讨脂多糖（LPS）对RAW264．7细胞白细胞介素10（interleuldn-10，IL-10）分泌及mRNA表达的影响。方法采用体外培养巨噬细胞株RAW264．7细胞，实验分为正常对照组、LPS组。采用ELISA法、逆转录PCR（RT—PCR）技术检测IIJ-10分泌及mRNA表达的变化。结果LPS刺激RAW264．7细胞后IL-10的分泌及mRNA表达较对照组增加（P〈0．05），并具有时间依赖性。结论LPS可刺激RAW264．7细胞IIJ—10分泌及mRNA表达的增加。     

AF-1对内毒素诱导RAW264．7细胞IL-10表达的影响

目的探讨抗炎素-1（Antiflammin-1,AF-1）对脂多糖（LPS）诱导的RAW264．7细胞白细胞介素10（interleuki-10,IL-10）表达的影响。方法采用体外培养巨噬细胞RAW264．7细胞,实验分为正常对照组,LPS组和不同浓度的LPS＋AF-1组。应用逆转录PCR（RT—PCR）技术检测IL-10表达的变化。结果1μg／ml的LPS刺激RAW264．7细胞后IL-10的表达较正常对照组增加（P〈0．05）,LPS＋AF-1组IL-10表达较LPS组表达显著升高（P〈0．05）,并具有剂量依赖性结论AF-1可促进LPS诱导的RAW264．7细胞IL-10表达的增加。     

绿原酸对脂多糖致炎症小鼠体内体外COX-2的影响

探讨绿原酸对脂多糖致炎症小鼠体内体外COX-2蛋白表达的影响.脂多糖滴鼻造模建立小鼠急性肺损伤模型,脂多糖诱导小鼠巨噬细胞RAW264.7建立细胞炎症反应模型,应用Weston blot法,检测小鼠肺组织内以及RAW264.7巨噬细胞中COX-2的蛋白表达.绿原酸在小鼠体内体外均能有效降低脂多糖诱导的COX-2的蛋白表达,并能促进COX-2蛋白的降解.降低脂多糖诱导的COX-2的蛋白水平是绿原酸抗炎的机制之一.     

两种石耳科多糖的理化特性及其抗炎症作用

为研究石耳科多糖的理化特性、微观结构和抗炎症作用,从石耳科两个相近物种石耳(Umbilicaria esculenta)和疱脐衣(Lasallia papulosa)中制备得到多糖CUEP和YUFP,采用高效液相色谱法、红外光谱法和透射电子显微镜分析两种多糖的化学成分和微观结构,利用脂多糖诱导RAW264.7细胞产生炎症因子,进而研究多糖对炎症因子mRNA相对表达量的影响.结果表明:从Umbilicaria esculenta和Lasallia papulosa中分离纯化得到CUEP和YUFP的得率分别为25%和27%,二者均含有糖类物质的特征官能团,CUEP由162~474 u的多糖链聚合而成,YUFP由98 u单一多糖组成;CUEP和YUFP均由葡萄糖、木糖、半乳糖、甘露糖组成,摩尔比分别为177.96∶1.00∶4.45∶3.19和33.21∶3.17∶1.00∶2.40;CUEP呈线状,糖链间相互缠绕且聚合度高、不易分散,而YUFP则呈蝴蝶结状,更易解聚;75 μg·mL^-1 YUFP 能显著地降低IL-1β的mRNA相对表达量,而75 μg·mL^-1 CUEP则能显著地降低炎症因子TNF-α,IL-1β,IL-6,COX-2的mRNA相对表达量.     

石松属植物化学成分抗炎活性的筛选

为研究2种石松属植物化学成分的抗炎活性,筛选具有较强抗炎活性的化合物,用LPS诱导RAW264.7细胞建立了体外炎症模型,采用MTT法检测了化合物对小鼠巨噬细胞株RAW264.7细胞增殖的影响.在安全浓度范围内(细胞存活率大于80%)给予药物干预,用Griess法检测了培养上清液中一氧化氮(NO)水平,以各化合物抑制NO释放的能力为筛选指标,评价了化合物的抗炎活性.结果表明:乙酰基二氢石松生物碱(16)、对香豆酸甲酯(21)、豆甾烷-3-酮-21-酸(22)具有较好的NO抑制率,并呈一定剂量依赖关系,提示它们具有一定的抗炎活性.     

基于灵芝双向固体发酵雷公藤减毒持效的研究

传统中药雷公藤具有抗炎和免疫抑制的作用,但因毒性强而极大地限制了其临床应用。以灵芝为药用真菌,采用双向固体发酵技术发酵雷公藤,并对发酵过程中得到的灵雷菌质进行抗炎活性和肝毒性评价。结果表明,接种量10 mL/100 mL、发酵30天得到的灵雷菌质（N2-G30）在3.75 μg/mL的质量浓度下表现出减毒持效的作用,相比于雷公藤,可抑制脂多糖诱导的小鼠RAW264.7细胞释放促炎因子TNF-α和IL-6,并且可减轻对人正常肝细胞L02增殖的抑制作用。通过液相色谱-质谱联用（LC-MS）分析雷公藤发酵前后主要成分的含量变化,结果显示,与发酵前相比,发酵后雷公藤甲素、雷公藤红素、雷酚内酯的含量增加,雷公藤内酯甲的含量减少,这些成分的含量变化与N2-G30的减毒持效作用有关。     

Vagus nerve stimulation attenuates the systemic inflammatory response to endotoxin

Vertebrates achieve internal homeostasis during infection or injury by balancing the activities of proinflammatory and anti-inflammatory pathways. Endotoxin (lipopolysaccharide), produced by all gram-negative bacteria, activates macrophages to release cytokines that are potentially lethal. The central nervous system regulates systemic inflammatory responses to endotoxin through humoral mechanisms. Activation of afferent vagus nerve fibres by endotoxin or cytokines stimulates hypothalamic-pituitary-adrenal anti-inflammatory responses. However, comparatively little is known about the role of efferent vagus nerve signalling in modulating inflammation. Here, we describe a previously unrecognized, parasympathetic anti-inflammatory pathway by which the brain modulates systemic inflammatory responses to endotoxin. Acetylcholine, the principle vagal neurotransmitter, significantly attenuated the release of cytokines (tumour necrosis factor (TNF), interleukin (IL)-1beta, IL-6 and IL-18), but not the anti-inflammatory cytokine IL-10, in lipopolysaccharide-stimulated human macrophage cultures. Direct electrical stimulation of the peripheral vagus nerve in vivo during lethal endotoxaemia in rats inhibited TNF synthesis in liver, attenuated peak serum TNF amounts, and prevented the development of shock.     

云芝多糖在NO诱导的巨噬细胞凋亡中的分子基础

在应用云芝多糖的基础上,用NO诱导居噬细胞凋亡,发现云芝多糖可加强NO诱导的细胞凋亡;通过RT－PCR观察诱导生性一氧化氮合酶基因的表达情况,结果表明云芝多糖能诱导一氧化氮合酶基因表达,并可增强γ干扰纱和脂多糖的诱导作用,提示云芝多糖在动脉粥样硬化中的防治作用是通过诱导诱生性一氧化氮合酶表达而实现的。     

茶叶糖蛋白对RAW264.7细胞分泌作用的影响

本文研究利用小鼠巨噬细胞建立体外实验模型,对茶叶糖蛋白(TGP)的免疫活性进行研究。采用倒置显微镜观察细胞形态变化。Griess法和ELISA法检测不同剂量茶叶糖蛋白对RAW264.7细胞一氧化氮(NO)和细胞因子TNF-α、IL-1β分泌量的影响。结果表明:50μg/mL茶叶糖蛋白可显著促进细胞分泌一氧化氮和细胞因子TNF-α、IL-1β(p<0.01),说明茶叶糖蛋白可通过促进细胞分泌一氧化氮和细胞因子,起到活化RAW264.7细胞和调节机体免疫的功效。     

Recognition and plasma clearance of endotoxin by scavenger receptors.

Lipid A is the active moiety of lipopolysaccharide (LPS, also referred to as endotoxin), a surface component of Gram-negative bacteria that stimulates macrophage activation and causes endotoxic shock. Macrophages can bind, internalize and partially degrade LPS, lipid A and its bioactive precursor, lipid IVA. We report here that lipid IVA binding and subsequent metabolism to a less active form by macrophage-like RAW 264.7 cells is mediated by the macrophage scavenger receptor. Scavenger-receptor ligands inhibit lipid IVA binding to, and metabolism by, RAW cells, and lipid IVA binds to type I and type II bovine scavenger receptors on transfected Chinese hamster ovary cells. Although in vitro competition studies with RAW cells indicate that scavenger receptor binding is not involved in LPS or lipid IVA-induced stimulation of macrophages, in vivo studies show that scavenger-receptor ligands greatly inhibit hepatic uptake of lipid IVA in mice. Thus, scavenger receptors expressed on macrophages may have an important role in the clearance and detoxification of endotoxin in animals.     

黄藤合剂的急性毒性和药效学研究

用热板法和扭体法研究黄藤合剂镇痛效果,测定毛细血管通透性和炎症组织PGE2含量以研究其抗炎活性及机理.小鼠灌胃给予黄藤合剂最大给药量76.8 g/kg后,7天内未见明显异常变化.黄藤合剂8mg/kg即能明显提高小鼠对热刺激的痛阈值,减少醋酸引起的扭体次数,降低毛细血管通透性,减少炎症组织PGE2的含量.16 g/kg,还能推迟疼痛反应出现的时间,延长潜伏期.黄藤合剂具有镇痛抗炎作用,最大给药量为76.8 g/kg,其作用机理可能与减少PGE2含量有关.     

参附注射液对内毒素休克大鼠肺组织表达PPARγ和TNFα的干预作用

摘要:目的 观察内毒素休克大鼠肺组织中过氧化物酶体增殖物激活受体γ(PPARγ)和肿瘤坏死因子α(TNFα)的表达情况及参附注射液的干预作用。方法 经腹腔注射脂多糖(LPS)建立内毒素休克SD大鼠模型,用低、中、高剂量参附注射液进行治疗,观察肺组织病L改变,检测肺组织中PPARγ和TNFα的表达量,同时检测血浆中TNFα和IL-1β的水平。结果 模型组大鼠具有典型的急性肺损伤表现;肺组织中PPARγ的转录和表达(P<0.01)量均显著下调,TNFα的表达明显增加(P<0.01);血浆中TNFα和IL-1β水平明显上升(P<0.01)。与模型组比较,参附注射液改善肺组织充血、水肿及炎性细胞浸润;呈剂量依赖性上调肺组织中PPARγ的转录和表达,下调TNFα的表达;同时抑制血浆中炎症介质TNFα和IL-1β水平(P<0.01)。结论 参附注射液保护内毒素休克大鼠肺组织,其作用机制可能与上调PPARγ从而抑制炎症介质的产生有关。     

Interleukin-2 mediates the peripheral analgesia of interferon alpha

Interferon-α (IFN-α) and interleukin-2 (IL-2) are crucial cytokines in immune system. They also play an important role in nerve system. It has been reported that IL-2 can induce the central analgesia. It is demonstrated that IFN-α also can induce the peripheral analgesia, which can be blocked by naloxone as IL-2. Furthermore, the analgesic effect of IFN-α is reversible by monoclonal anti-IL-2 antibody. In vitro experiments show that IFN-α significantly increases the production of IL-2 in a dose dependent manner. These data suggest that IL-2 mediates the peripheral analgesia of IFN-α.     

氢吗啡酮对炎性痛大鼠脊髓ERK1/2信号通路的影响

目的 探讨氢吗啡酮对炎性痛大鼠脊髓ERK1/2信号通路的影响.方法 选择SPF级雄性大鼠30只,随机分为3组,正常组、模型组和氢吗啡酮组;检测各组大鼠热痛阈和机械痛阈;ELISA检测血清中TNF-α、IL-6和IL-10的含量;Western blot检测各组大鼠脊髓中ERK1/2和p-ERK1/2蛋白表达水平;qRT...     

1-磷酸鞘氨醇受体1促进心肌梗死后单核/巨噬细胞在梗死心肌组织中浸润和聚集

目的探讨心肌梗死后S1PR1对单核/巨噬细胞功能的影响及作用机制。方法构建急性心肌梗死模型;实验分为实验组(S1PR1激动剂SEW2871处理组)、阴性对照组(DMSO处理组),各组小鼠分别在药物处理后0、5、28 d三个时间点进行对比研究;流式细胞术检测心肌组织、肝脏、肾脏及外周血等组织中单核/巨噬细胞的数量;荧光显微镜下观察心肌组织单核/巨噬细胞荧光表达情况;构建小鼠pCMV.DR8和pMD2.G慢病毒载体并感染小鼠RAW264.7巨噬细胞,筛选最适感染复数(MOI);实验分为S1PR1基因沉默组、过表达组、U0126(ERK信号通路阻断剂)处理组及空白对照组;采用Transwell小室分析细胞迁移情况;通过RAW264.7与HUVECSs共培养检测细胞粘附功能。结果 (1)实验组小鼠心肌组织中单核/巨噬细胞数量明显多于对照组(P 0.05);(2)体外试验显示,SEW2871促进RAW264.7巨噬细胞的粘附和迁移,S1PR1基因敲减后,上述作用明显降低;(3) U0126预处理后,SEW2871的促进细胞粘附和迁移作用显著减弱。结论 S1PR1通过ERK信号通路促进单核/巨噬细胞的粘附和迁移作用。     

Pathogen-induced human TH17 cells produce IFN-γ or IL-10 and are regulated by IL-1β

IL-17-producing CD4+ T helper cells (TH17) have been extensively investigated in mouse models of autoimmunity. However, the requirements for differentiation and the properties of pathogen-induced human TH17 cells remain poorly defined. Using an approach that combines the in vitro priming of naive T cells with the ex vivo analysis of memory T cells, we describe here two types of human TH17 cells with distinct effector function and differentiation requirements. Candida albicans-specific TH17 cells produced IL-17 and IFN-γ, but no IL-10, whereas Staphylococcus aureus-specific TH17 cells produced IL-17 and could produce IL-10 upon restimulation. IL-6, IL-23 and IL-1β contributed to TH17 differentiation induced by both pathogens, but IL-1β was essential in C. albicans-induced TH17 differentiation to counteract the inhibitory activity of IL-12 and to prime IL-17/IFN-γ double-producing cells. In addition, IL-1β inhibited IL-10 production in differentiating and in memory TH17 cells, whereas blockade of IL-1β in vivo led to increased IL-10 production by memory TH17 cells. We also show that, after restimulation, TH17 cells transiently downregulated IL-17 production through a mechanism that involved IL-2-induced activation of STAT5 and decreased expression of ROR-γt. Taken together these findings demonstrate that by eliciting different cytokines C. albicans and S. aureus prime TH17 cells that produce either IFN-γ or IL-10, and identify IL-1β and IL-2 as pro- and anti-inflammatory regulators of TH17 cells both at priming and in the effector phase.     

三峡区域特色药材宜昌润楠正丁醇提取物体外抗炎活性研究

目的：探讨宜昌润楠正丁醇提取物（MCB）的体外抗炎活性．方法：利用细菌脂多糖（LPS）刺激经佛波酯（PMA）诱导的人单核细胞THP-1建立体外炎症模型,MTT法检测样品的细胞毒性,ELISA方法检测药物干预前后细胞上清液中IL-1β、IL-6、TNF—α和NO的分泌量,综合评价MCB对炎性细胞因子和介质的抑制作用．结果：MCB显著性地抑制了IL-6和TNF—a以及NO的产生,并且其抑制作用具有时间和剂量依耐性,但对IL-1β没有明显的抑制效果．结论：本研究初步验证了宜昌润楠的体外抗炎作用,为其民间应用提供了理论依据,也为进-步分离生物活性物质提供了方向．     

Intravenous gammaglobulin suppresses inflammation through a novel T(H)2 pathway

High-dose intravenous immunoglobulin is a widely used therapeutic preparation of highly purified immunoglobulin G (IgG) antibodies. It is administered at high doses (1-2 grams per kilogram) for the suppression of autoantibody-triggered inflammation in a variety of clinical settings. This anti-inflammatory activity of intravenous immunoglobulin is triggered by a minor population of IgG crystallizable fragments (Fcs), with glycans terminating in α2,6 sialic acids (sFc) that target myeloid regulatory cells expressing the lectin dendritic-cell-specific ICAM-3 grabbing non-integrin (DC-SIGN; also known as CD209). Here, to characterize this response in detail, we generated humanized DC-SIGN mice (hDC-SIGN), and demonstrate that the anti-inflammatory activity of intravenous immunoglobulin can be recapitulated by the transfer of bone-marrow-derived sFc-treated hDC-SIGN(+) macrophages or dendritic cells into naive recipients. Furthermore, sFc administration results in the production of IL-33, which, in turn, induces expansion of IL-4-producing basophils that promote increased expression of the inhibitory Fc receptor FcγRIIB on effector macrophages. Systemic administration of the T(H)2 cytokines IL-33 or IL-4 upregulates FcγRIIB on macrophages, and suppresses serum-induced arthritis. Consistent with these results, transfer of IL-33-treated basophils suppressed induced arthritic inflammation. This novel DC-SIGN-T(H)2 pathway initiated by an endogenous ligand, sFc, provides an intrinsic mechanism for maintaining immune homeostasis that could be manipulated to provide therapeutic benefit in autoimmune diseases.