松杉灵芝菌丝体多糖(FI_0-b)对人组织瘤细胞和人外周血白细胞产生炎症性细胞因子的影响(Ⅰ)(英文)

目的:比较研究松杉灵芝水溶性多糖(Fl_0-b)及其甲酸修饰产物(FI_0-b-H)对人炎症性细胞因子产生的影响.方法:用放射免疫分析法(RIA)及逆转录聚合酶链反应(RT-PCR)分析法研究FI_0-b和Fl_0-b-H对人组织瘤细胞(THP-1)和人外周血单核细胞(PBMC)分泌的各种与炎症有关的细胞因子(白介素-1 α,6,8,肿瘤坏死因子α)及对白介素-1α,肿瘤坏死因子αmRNA表达的影响.结果:FI_0-b和FI_0-b-H(浓度分别为4,40,400 mg/L)显著抑制LPS 10 mg/L协同PMA 200 nmol/L诱导的THP-1细胞和PBMC细胞产生IL-1α和TNF α的量.当低浓度刺激剂(LPS 1mg/L协同PMA 200 nmol/L)刺激THP-1细胞时,FI_0-b-H明显提高白介素-8的产生;但当高浓度刺激剂(LPS 10 mg/L协同200 nmol/L PMA)刺激THP-1-细胞时,FI_0-b-H却明显抑制白介素-8的产生.FI_0-b-和FI_0-b-H对THP-1细胞白介素-1α和肿瘤坏死因子α的mRNA表达有显著作用.结论:与FI_0-b相比,FI_0-b-H在人炎症性细胞因子的产生方面表现出较强的活性.松杉灵芝菌丝体水溶性多糖在不同的刺激条件下,具有双向免疫调节作用,并具有一定的剂量依赖关系.     

铁杉灵芝多糖FⅢ-2对人组织瘤细胞和人外周血白细胞产生炎症性细胞因子的影响(英文)

用放射免疫分析法及逆转录聚合酶链反应法 ,比较研究水不溶性铁杉灵芝多糖 (FⅢ 2 )及其吡啶 氯磺酸修饰产物 (FⅢ 2 S)对人炎症性细胞因子蛋白质及其mRNA产生的影响 .结果表明 ,FⅢ 2 (4 ,4 0或 4 0 0mg·L- 1)显著提高低剂量脂多糖 (LPS) 10mg·L- 1协同佛波醇 14烷酰乙酸盐 (PMA) 2 0 0nmo1·L- 1诱导的人组织瘤THP 1细胞产生肿瘤坏死因子α(TNFα) ,然而明显地抑制高剂量LPSl0 0mg·L- 1协同PMA诱导的THP 1细胞产生TNFα .FⅢ 2 S(4或4 0mg·L- 1)提高无刺激剂细胞产生TNFα ,高浓度FⅢ 2 S(4 0 0mg·L- 1)抑制TNFα产生 .FⅢ 2与FⅢ 2 S提高白介素 8的产生 ,降低刺激剂引起的白介素 1α的产生 .FⅢ 2的抗肿瘤作用可能是与TNFα产生有关 .FⅢ 2对人外周血单核细胞产生各种细胞因子和对THP 1细胞产生细胞因子的机理基本上相同 .结果提示 ,松杉灵芝菌丝体水不溶性多糖在不同的刺激条件下具有双向免疫调节作用 .化学修饰的多糖可改变原多糖对细胞因子产生的调节方向 .     

人参皂苷Rg_1的肠内菌代谢Ⅱ 人参皂苷Rg_1和Rh_1免疫活性(英文)

目的:比较人参皂苷Rg_1及其代谢产物Rh_1对细胞因子及其mRNA表达的影响.方法:将Rg_1及Rh_1加入正常人外周血单核细胞(PBMC)培养24小时后,计数细胞,观察其对正常细胞增殖的影响.用放射免疫法(RIA)观察Rg_1及Rh_1对人组织瘤细胞(THP-1)分泌与炎症有关的细胞因子(IL-1α,TNFα,IL-8)产生的影响,用逆转录酶链式聚合反应(RT-PCR)方法,检测Rg_1及Rh_1对TNFα的mRNA表达的影响.结果:Rg_1及Rh_1(0.1,1,10,100 mg/L)对PBMC增殖无明显影响.但在脂多糖10 mg/L和PMA 200 nmol/L存在下,Rh_1的低浓度(1 mg/L)能促进THP-1细胞产生TNFα与IL-8.而Rg_1在LPS 100 mg/L时抑制TNFα的产生.Rg_1 1 mg/L及Rh_1 100 mg/L均能促进IL-1α的产生.RT-PCR实验结果表明,Rh_1能显著促进TNFα mRNA的表达.结论:Rg_1与Rh_1的免疫活性有所不同,在有些方面甚至相反.     

淫羊藿苷及其肠菌代谢产物对THP-1细胞分泌细胞因子的影响

目的：比较淫羊藿苷及其肠菌代谢产物对人组织细胞瘤THP-1细胞分泌4种细胞因子的影响。方法：放射免疫测定方法(RIA)。 结果：在LPS和PMA存在下, 淫羊藿苷及其肠菌代谢产物(宝藿苷I和淫羊藿苷元）对IL-6的产生有促进作用, 代谢物的作用更强, 对IL-8的产生有一定的抑制作用。当LPS和PMA不存在时,对IL-6的产生无明显影响, 而对IL-8的产生则有促进作用。 不论LPS和PMA存在与否,原苷对TNFα的产生有抑制作用, 原苷及其肠菌代谢产物对IL-1α的产生均有明显抑制作用。结论：在不同的实验条件下,淫羊藿苷及其肠菌代谢产物对各种炎症性细胞因子的产生均有特异的调节作用。     

雷公藤氯内酯醇对小鼠肺泡巨噬细胞体外炎性反应的影响(英文)

目的:探讨雷公藤单体雷公藤氯内酯醇(tripcholorolide,T4)对肺泡巨噬细胞(AM)炎症反应的影响及机制.方法:小鼠AM受脂多糖(LPS)10mg/L刺激的同时,加入T4 500 μg/L或地塞米松 100μmol/L;ELISA法测定上清液中TNFα、IL-1β、IL-6及IL-10浓度:RT-PCR检测上述因子及iNOS基因mRNA的表达.结果:AM受10 mg/L LPS刺激24小时后,上清液中TNFα、IL-1β、IL-6、IL-10及NO释放均明显增加.T4 500 μg/L及地塞米松100μmol/L对上述介质均有不同程度的抑制作用.LPS刺激5小时后,AM中TNFα、IL-6、IL-10和iNOS的mRNA表达均明显增加.T4和地塞米松对上述介质的mRNA表达均有明显抑制作用.另外,T4对TNFα、IL-6、IL-10 mRNA的稳定性无明显影响.结论:T4具有抑制AM中促炎介质和抗炎介质表达的作用.     

复方黄芪提取物对细菌脂多糖诱导大鼠腹腔巨噬细胞分泌TNFα、NO及IL-1的影响

目的研究复方黄芪提取物(CAE)对细菌脂多糖诱导大鼠腹腔巨噬细胞(PMΦs)释放肿瘤坏死因子((TNFα)、白细胞介素-1(IL-1)及一氧化氮(NO)的影响。方法10 mg/L LPS体外诱导大鼠PMΦs分泌TNFα、IL-1与NO等因子,采用小鼠成纤维细胞瘤株(L929)杀伤法检测TNFα的含量、小鼠胸腺细胞增殖法检测IL-1的活性、G riess法检测NO的水平。结果CAE(11.3~90 mg.L-1)能显著抑制LPS激活的巨噬细胞对TNFα、IL-1与NO的分泌。结论CAE抑制巨噬细胞的分泌功能可能是其保肝作用的机制之一。     

油酰乙醇胺对脂多糖诱导的THP-1细胞炎症因子表达的影响

目的探讨油酰乙醇胺(OEA)对细菌脂多糖(LPS)诱导的人急性白血病单核细胞(THP-1)中前炎症因子TNF-α、IL-1β、IL-6表达的影响,并初步探讨OEA作为过氧化物酶体增殖物激活受体-α(PPAR-α)激动剂参与对炎症调节的作用机制。方法体外培养的THP-1细胞,分别加入不同浓度的OEA(10,20,40μmol/L)或非诺贝特(100μmol/L)共同孵育1 h后,用1μg/mL LPS分别诱导6或24 h。采用RT-PCR、实时定量PCR和酶联免疫吸附检测测定细胞中TNF-α、IL-1β、IL-6 mRNA和蛋白的表达的变化,并使用实时定量PCR及Western blot方法检测PPAR-α及Toll样受体4(TLR4)的mRNA和蛋白的表达。结果相对于正常THP-1细胞,LPS诱导后细胞中炎症因子(TNF-α、IL-1β、IL-6)表达明显增加。OEA对TNF-α、IL-1β、IL-6 mRNA和蛋白的表达有抑制作用,并呈现出一定的剂量依赖性。且OEA在激活PPAR-α表达的同时能够抑制TLR4的表达。结论 OEA对LPS诱导的炎症反应有抑制作用,其机制可能与激活PPAR-α,下调TLR4的表达有关。     

AP对正常和内毒素化大鼠脾细胞体外分泌细胞因子的影响

从分子免疫水平探讨了巴巴多斯芦荟多糖提取物(AP)对免疫系统的作用.结果表明巴巴多斯芦荟多糖提取物对体外正常大鼠脾细胞分泌IL-1、IL-6、TNFα等细胞因子均有一定的促进作用,而对内毒素诱导脾细胞分泌的细胞因子IL-1、IL-6、TNFα有显著抑制作用.对IL-2的分泌在以上两种情况下均表现促进作用.显示巴巴多斯芦荟多糖提取物对细胞因子的分泌可产生双向调控作用.     

异丙酚抑制内质网应激减轻脂多糖诱导人肝细胞损伤的机制研究

目的 探讨异丙酚(PPF)对脂多糖(LPS)诱导人正常肝细胞L-02损伤的保护作用.方法 用不同浓度的LPS来制备人肝细胞损伤模型,将人正常肝细胞L-02按随机数字表法分为五组:正常组、LPS组、LPS+25、50μmol/L PPF组以及LPS+4 mmol/L 4-苯基丁酸(PBA)组.四甲基偶氮唑盐微量酶反应比色法(MTT法)检测L-02细胞的存活率,酶联免疫吸附测定(ELISA)测定各组细胞上清中丙氨酸氨基转移酶(ALT)和天冬氨酸氨基转移酶(AST)的活性.实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测各组细胞中炎性因子白细胞介素(IL)-1β、IL-6和肿瘤坏死因子α(TNF-α)mRNA的表达;蛋白质印迹法(Western blotting)检测各组细胞中核因子-κB(NF-κB)p65、NF-κB、p-肌醇依赖酶1α(IRE1α)、IRE1α和激活转录因子6(ATF-6)蛋白的表达.结果 用不同浓度的LPS处理的人肝细胞存活能力显著降低,且LPS致人肝细胞损伤的最佳造模条件为20 mg/L孵育24 h.与正常组比较,LPS组的人肝细胞存活能力显著降低,细胞数量减少且核固缩;细胞上清中ALT和AST的释放量分别增加[(69.17±5.51)mg/L和(50.23±5.81)mg/L比(19.78±1.79)mg/L和(9.79±0.96)mg/L,P<0.05];IL-1β、IL-6和TNF-αmRNA表达显著升高;NF-κB p65表达量明显增强,且内质网应激相关蛋白p-IRE1α/IRE1α和ATF-6的表达显著增高[(2.24±0.28)和(2.55±0.16)比(1.00±0.17)和(1.00±0.26),P<0.05].与LPS组相比,给予低高剂量PPF能显著提高人肝细胞的存活率,降低ALT[(55.30±6.92)mg/L和(33.70±4.08)mg/L]和AST[(39.46±2.56)mg/L和(27.62±2.12)mg/L]的释放量;显著降低IL-1β、IL-6和TNF-αmRNA的表达;抑制NF-κB p65及内质网应激相关蛋白ATF-6和p-IRE1α/IRE1α蛋白水平的表达.结论 PPF对LPS诱导的人肝细胞损伤具有一定的保护作用,可能是通过抑制内质网应激从而发挥抗炎作用来实现的.     

壬二酸和水杨酸对痤疮丙酸杆菌诱导的细胞炎症因子及TLR4蛋白表达的影响

目的 探讨壬二酸和水杨酸对痤疮丙酸杆菌Propionibacterium acnes诱导的炎症因子及Toll样受体4(TLR4)表达的影响。方法 采用CCK-8法检测药物对人角质形成细胞HaCaT和人单核细胞THP-1增殖活力的影响,酶联免疫吸附法（ELISA）检测细胞培养上清中白细胞介素（IL）-8、IL-1β和肿瘤坏死因子-α(TNF-α)浓度,利用分子对接技术分析壬二酸、水杨酸与TLR4蛋白的分子间相互作用,并通过细胞免疫荧光和荧光强度分析验证其对Ha Ca T细胞表面TLR4蛋白表达的影响。结果 经不同浓度的壬二酸或水杨酸处理48 h后,在Ha CaT细胞实验中,壬二酸的最大实验质量浓度为62.50μg/mL,水杨酸为31.25μg/mL;THP-1细胞实验中壬二酸的最大实验质量浓度为1 000.00μg/m L,水杨酸为500.00μg/m L。壬二酸和水杨酸对P. acnes胞壁成分肽聚糖（PGN）+脂磷壁酸（LTA）或热灭活P. acnes诱导后HaCaT细胞产生IL-8以及对佛波酯（PMA）+细菌脂多糖（LPS）或热灭活P. acnes诱导后的THP-1细胞产生IL-1...     

Interleukin-6 can prime THP-1 macrophages for enhanced production of tumor necrosis factor-alpha in response to LPS.

Although interferon-gamma has been shown to effectively prime macrophages for enhanced production of tumor necrosis factor-alpha (TNF alpha), it is reasonable to assume that other cytokines present in the extracellular environment may likewise facilitate cytokine biosynthesis. For example, interleukin-6 (IL-6) is synthesized by synovial lining macrophages and fibroblasts, and has been detected (along with TNF alpha) in rheumatoid synovial effusions. Therefore, the purpose of the present study was to determine whether IL-6 influences the production of IL-1 beta and/or TNF alpha by THP-1 macrophages. Although IL-6 treatment alone resulted in only a slight increase in TNF alpha levels, administration of IL-6 followed by Sal. minnesota LPS resulted in a synergistic potentiation of TNF alpha production by THP-1 macrophages. The priming effect of IL-6 could be reversed by boiling, or by the addition of a neutralizing polyclonal antibody against IL-6. Notably, IL-6 only weakly enhanced interleukin-1 beta production. In summary, the ability of IL-6 to potentiate TNF alpha production by THP-1 macrophages may provide insight into the regulation of the cytokine network in inflammatory diseases, such as rheumatoid arthritis.     

Regulation of interleukin-1 beta and tumor necrosis factor secretion by the human monocytic leukemia cell line, THP-1

Activated macrophages synthesize and release the potent polypeptides, interleukin-1 (IL-1) and tumor necrosis factor (TNF). In an effort to identify the cellular signals which control cytokine production by activated macrophages, we have developed an in vitro model employing the human THP-1 cell line. In the present study, THP-1 cells "primed" by 1.6 microM phorbol 12-myristate-13-acetate (TPA) for 4 hr demonstrated a dose- and time-dependent release of IL-1 beta and TNF upon activation by 20 micrograms/ml LPS. BSA/anti-BSA-coated latex beads were also a potent stimulus for IL-1 beta secretion. Moreover, the combination of a suboptimal concentration of LPS (200 ng/ml) plus interferon-gamma (0.03-333 U/ml) greatly enhanced IL-1 beta production. Resting THP-1 monocytes not "primed" by TPA did not secrete IL-1 beta or TNF. These distinct patterns of cytokine production may be related to the developmental stages of macrophage activation.     

Decursin inhibits induction of inflammatory mediators by blocking nuclear factor-kappaB activation in macrophages

In the course of screening inhibitors of matrix metalloproteinase (MMP)-9 induction in macrophages, we isolated decursin, a coumarin compound, from the roots of Angelicae gigas. As a marker for the screening and isolation, we tested expression of MMP-9 in RAW264.7 cells and THP-1 cells after treatment with bacterial lipopolysaccharide (LPS), the TLR-4 ligand. Decursin suppressed MMP-9 expression in cells stimulated by LPS in a dose-dependent manner at concentrations below 60 microM with no sign of cytotoxicity. The suppressive effect of decursin was observed not only in cells stimulated with ligands for TLR4, TLR2, TLR3, and TLR9 but also in cells stimulated with interleukin (IL)-1beta, and tumor necrosis factor (TNF)-alpha, indicating that the molecular target of decursin is common signaling molecules induced by these stimulants. In addition to the suppression of MMP-9 expression, decursin blocked nitric oxide production and cytokine (IL-8, MCP-1, IL-1beta, and TNF-alpha) secretion induced by LPS. To find out the molecular mechanism responsible for the suppressive effect of decursin, we analyzed signaling molecules involved in the TLR-mediated activation of MMP-9 and cytokines. Decursin blocked phosphorylation of IkappaB and nuclear translocation of NF-kappaB in THP-1 cells activated with LPS. Furthermore, expression of a luciferase reporter gene under the promoter containing NF-kappaB binding sites was blocked by decursin. These data indicate that decursin is a novel inhibitor of NF-kappaB activation in signaling induced by TLR ligands and cytokines.