来氟米特对佐剂性关节炎大鼠腹腔巨噬细胞IL—1，IL—6和TNF—α产生的动态影响

目的:探讨来氟米特(leflunomide,LEF)对佐剂性关节炎大鼠腹腔巨噬细胞IL-1,IL-6和 TNF-α分泌的影响及其抗炎、抗类风湿的可能作用机制.方法:大鼠足跖皮下注射Freund完全佐剂诱导关节炎模型;LEF灌胃后分次获取腹腔巨噬细胞,其培养上清液中IL-1,IL-6和TNF-α活性采用ELISA法或生物法测定.结果:佐剂性关节炎大鼠腹腔巨噬细胞IL-1,IL-6和TNF-α分泌较正常对照组明显升高;LEF对由 LPS诱导产生的 IL-1和 TNF-α有明显的抑制作用,作用产生快;LEF(10,25 mg/kg)在应用21天后对IL-6的分泌也有明显抑制作用.结论:来氟米特具有抑制佐剂性关节炎大鼠腹腔巨噬细胞 IL-1,IL-6和 TNF-α分泌水平的作用.     

匹多莫德对免疫功能缺陷小鼠脾组织TNF-α和IL-6表达及相关免疫药理作用的研究

目的:研究匹多莫德注射剂对正常小鼠及免疫功能低下小鼠脾组织TNF-α和IL-6表达及相关免疫功能的影响,以及匹多莫德免疫调节作用可能的机制。方法:RT-PCR法对小鼠脾组织的TNF-α和IL-6的表达进行分析;中性红法对小鼠的腹腔巨噬细胞的吞噬功能进行研究;MTT法检测ConA和LPS分别刺激的小鼠脾淋巴细胞的体外增殖;血清溶血素法检测小鼠血清抗体的产生。结果:匹多莫德注射剂连续给药14d对正常小鼠及免疫功能低下小鼠脾组织的TNF-α和IL-6的表达有一定的促进作用;小鼠腹腔巨噬细胞的吞噬能力明显提高;ConA或LPS刺激的小鼠脾淋巴细胞的转化能力均有明显的提高。结论:匹多莫德可明显提高正常小鼠和免疫功能低下小鼠的免疫功能,并促进TNF-α和IL-6表达。     

黄芪皂甙Ⅳ对小鼠T、B淋巴细胞增殖和腹腔巨噬细胞功能的影响(英文)

目的:探讨黄芪皂苷Ⅳ(ASI)对小鼠T、B淋巴细胞增殖和腹腔巨噬细胞分泌的细胞因子的影响.方法:采用MTT法检测T、B淋巴细胞的增殖;采用分光光度计测定法检测抗体活性;IL-1活性用胸腺细胞增殖法测定;TNF-α活性用L929细胞杀伤法测定.结果:1)ASI50-200mg/kg ig 7天能够促进T淋巴细胞增殖和抗体生成,而ASI 50-100mg/kg能够促进B淋巴细胞增殖,但是220mg/kg对B淋巴细胞增殖无影响;2)ASI体外仅在100nmol/L对T、B淋巴细胞有促进作用;3)ASI 1 nmol/L可以促进腹腔巨噬细胞分泌IL-1,而100-1000nmol/L则抑制腹腔巨噬细胞分泌IL-1;4)ASI体外可以抑制LPS刺激或无LPS刺激下的腹腔巨噬细胞分泌TNF-α.结论:ASI能够促进小鼠T、B淋巴细胞的增殖和抗体生成,同时可以抑制腹腔巨噬细胞体外分泌IL-1和TNF-α.     

灵芝多糖/硒化卡拉胶口服液对免疫抑制小鼠免疫功能的影响

目的:研究灵芝多糖/硒化卡拉胶口服液对环磷酰胺诱导免疫抑制小鼠的非特异性免疫功能、体液免疫功能、细胞免疫功能的影响。方法:小鼠腹腔注射80mg/kg环磷酰胺诱导免疫抑制小鼠模型,灵芝多糖/硒化卡拉胶口服液连续供应30d,每天1次。测定小鼠单核巨噬细胞吞噬功能、腹腔巨噬细胞吞噬鸡红细胞功能、外周血白细胞数目、NK细胞活性、白介素-1(IL-1)活性、白介素-2(IL-2)活性、TNF-α活性、半数血清溶血素、抗体生成细胞、T、B淋巴细胞增殖能力、迟发型变态反应。结果:灵芝多糖/硒化卡拉胶口服液各剂量组(低剂量组:灵芝多糖2.5mg/kg+硒5μg/kg;中剂量组:灵芝多糖5mg/kg+硒10μg/kg;高剂量组:灵芝多糖15mg/kg+硒30μg/kg)均可提高外周血白细胞数目、提高半数溶血素值,低剂量组增强IL-1活性,中剂量组增强IL-2活性,高剂量组和中剂量组增强T、B淋巴细胞的增殖能力、NK细胞活性、TNF-α活性,高剂量组增强腹腔巨噬细胞吞噬功能、碳粒廓清值、提高抗体生成细胞、增强迟发型变态反应。结论:灵芝多糖/硒化卡拉胶口服液对免疫抑制小鼠的免疫功能具有改善作用。     

玉郎伞多糖对体外BALB/C小鼠淋巴细胞和巨噬细胞分泌细胞因子的影响

目的:探讨玉郎伞多糖（YLSPS）体外对BALB/C小鼠淋巴细胞、巨噬细胞分泌肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）、γ-干扰素（IFN-γ）及白细胞介素-2（IL-2）的影响及作用机制。方法:分离BALB/C小鼠腹腔巨噬细胞和脾淋巴细胞,分为空白对照组、LPS或ConA对照组和各浓度YLSPS组(50,100,200 mg·L^-1),MTT法检测脾淋巴细胞增殖,ELISA法检测细胞培养液中TNF-α、IL-6、IFN-γ、IL-2的浓度,RT-PCR法检测TNF-α、IL-6、IFN-γ、IL-2 mRNA的表达。结果:各浓度YLSPS组能剂量依赖性地促进BALB/C小鼠脾淋巴细胞增殖和TNF-α、IL-6、IFN-γ、IL-2的分泌,显著增强这些细胞因子的mRNA的表达（P<0.01或P<0.05）。结论:YLSPS体外能促进BALB/C小鼠脾淋巴细胞和腹腔巨噬细胞分泌IFN-γ、IL-2、TNF-α和IL-6,其作用机制可能与上调上述细胞因子mRNA表达有关。     

肉苁蓉多糖的巨噬细胞活化作用

目的观察肉苁蓉多糖(CDPS)对巨噬细胞的活化作用。方法采用中性红法测定吞噬活性;Griess试剂法测定NO释放量;L929细胞法及小鼠胸腺细胞法测定TNF-α及IL-1释放。结果CDPS在6.25～50mg.L-1剂量范围内可剂量依赖性地增强BALB/c小鼠腹腔巨噬细胞吞噬中性红能力和促进NO释放;在2.8～100mg.L-1剂量范围内可使正常和免疫抑制的RAW264.7细胞NO释放明显增加;在0.56～7.2mg.L-1或3.6～10mg.L-1范围促进RAW264.7细胞TNF-α或IL-1产生,均具有良好的量效关系。结论CDPS可明显提高巨噬细胞吞噬及分泌功能,活化巨噬细胞。肉苁蓉免疫增强作用可能与此有关。     

枸杞糖缀合物和糖链对小鼠巨噬细胞功能的影响

目的确定巨噬细胞是枸杞糖缀合物和糖链免疫作用的靶细胞之一。方法应用中性红吞噬实验和鸡红细胞吞噬实验测定了枸杞糖缀合物LbGp4和糖链LbGp4-OL对小鼠腹腔巨噬细胞吞噬功能的影响;用硝酸根还原法、酶联免疫吸附实验(ELISA)和生物活性测定法测定了巨噬细胞产生一氧化氮(NO)、IL-1β和TNF-α含量和生物活性的变化。结果LbGp4和LbGp4-OL在(10~100)mg.L-1剂量范围内均可剂量依赖性地促进静息巨噬细胞吞噬中性红的能力,增加活化的巨噬细胞吞噬鸡红细胞的吞噬率和吞噬指数;增加巨噬细胞培养上清NO、IL-1β和TNF-α的浓度,并增强对L929细胞的杀伤活性,促进胸腺细胞的增殖反应。结论LbGp4和LbGp4-OL对静息和活化的腹腔巨噬细胞的吞噬功能具有明显的促进作用,对巨噬细胞产生NO、分泌IL-1β和TNF-α的含量和生物活性亦具有明显的促进作用,表明巨噬细胞是LbGp4和LbGp4-OL免疫作用的靶细胞之一。     

仿刺参糖胺聚糖对小鼠腹腔巨噬细胞功能的影响

目的 研究仿刺参糖胺聚糖（HGAG）对小鼠腹腔巨噬细胞（Mφ）功能的影响,探讨其对小鼠的免疫调节作用。方法 不同浓度HGAG作用于体外培养Mφ,MTT比色法测定代谢活力;中性红法测定吞噬功能;试剂盒测定Mφ中乳酸脱氢酶（LDH）活性;Griess法测定NO生成量;ELISA法测定培养上清中TNF-α、IL-1β的分泌水平。结果 HGAG在0.1~25 μg·mL?1范围内对Mφ的功能有一定的促进作用。在1~10 μg·mL?1范围内与空白对照组相比可显著增强Mφ代谢活力和吞噬能力（P<0.01）,提高LDH活力和NO生成量（P<0.05）,显著增加培养上清中TNF-α和IL-1β（P<0.01）的浓度。与阳性对照组相比,在1~10 μg·mL?1范围内对Mφ吞噬功能以及TNF-α、IL-1β的生成有更好的促进作用（P<0.05）,在高浓度（100μ g·mL?1）Mφ的各项功能指标均低于空白对照组。结论 HGAG在1~10 μg·mL?1浓度范围内可显著促进Mφ活性,并在一定程度上优于阳性对照组。表明HGAG能够激活Mφ,促进Mφ的功能,增强机体免疫。     

无花果多糖对免疫抑制小鼠腹腔巨噬细胞产生IL-1α、脾细胞体外增殖、脾细胞产生IL-2及其受体的影响

目的　探讨无花果多糖对小鼠腹腔巨噬细胞产生IL-1α、脾细胞体外增殖、脾细胞产生和分泌白细胞介素2(IL-2)和血清可溶性白细胞介素2受体(SIL-2R) 水平的影响。方法　以环磷酰胺致免疫低下小鼠为研究对象,分为大、小剂量(剂量分别为400,200 mg·kg^-1)的无花果多糖水溶液组,香菇多糖组(100 mg·kg^-1)及同体积生理盐水组,另设空白对照组,每组6只。造模同时给药,连续7 d,测定小鼠腹腔巨噬细胞IL-1α、脾细胞体外增殖、脾细胞IL-2及血清SIL-2R水平。结果　无花果多糖可促进免疫抑制小鼠腹腔巨噬细胞产生和分泌IL-1α,脾细胞产生和分泌IL-2,促进ConA和LPS刺激的脾细胞增殖,降低血清SIL-2R水平。结论　无花果多糖有较好的免疫增强作用。     

无花果多糖对免疫抑制小鼠腹腔巨噬细胞产生IL-1α、脾细胞体外增殖、脾细胞产生IL-2及其受体的影响

目的探讨无花果多糖对小鼠腹腔巨噬细胞产生IL-1α、脾细胞体外增殖、脾细胞产生和分泌白细胞介素2（IL-2）和血清可溶性白细胞介素2受体（SIL-2R）水平的影响。方法以环磷酰胺致免疫低下小鼠为研究对象,分为大、小剂量（剂量分别为400,200mg·kg^-1）的无花果多糖水溶液组,香菇多糖组（100mg·kg^-1）及同体积生理盐水组,另设空白对照组,每组6只。造模同时给药,连续7d,测定小鼠腹腔巨噬细胞IL-1α、脾细胞体外增殖、脾细胞IL-2及血清SIL-2R水平。结果无花果多糖可促进免疫抑制小鼠腹腔巨噬细胞产生和分泌IL-1α,脾细胞产生和分泌IL-2,促进ConA和LPS刺激的脾细胞增殖,降低血清SIL-2R水平。结论无花果多糖有较好的免疫增强作用。     

Toll-like receptor 4-mediated ROS signaling pathway involved in Ganoderma atrum polysaccharide-induced tumor necrosis factor-α secretion during macrophage activation

Ganoderma atrum has been used as Chinese traditional medicine and healthful mushroom for thousands of years. The polysaccharide is regarded as the major bioactive substances in G. atrum. To delineate the underlying mechanism and signaling cascade involved in the immunomodulatory property of G. atrum polysaccharide (PSG-1). Specifically, this study is designed to examine the possibility of TLR4 as a candidate receptor interacted with G. atrum polysaccharide (PSG-1) and elucidate the role of reactive oxygen species (ROS) in PSG-1-induced tumor necrosis factor-α (TNF-α) production during macrophage activation. Flow cytometric and confocal laser-scanning microscopy analysis showed that fluorescence-labeled PSG-1 bind specifically to the macrophages. Moreover, PSG-1 stimulated TNF-α secretion of peritoneal macrophages from C3H/HeN mice, but not from C3H/HeJ mice. PSG-1-indcued TNF-α production was suppressed by anti-TLR4 mAb. Furthermore, ROS production was mediated by TLR4, and NADPH oxidase-derived ROS act as upstream of phosphoinositide 3-kinase(PI3K)/Akt/mitogen-activated protein kinases(MAPKs)/nuclear factor(NF)-κB signaling pathway in the regulation of PSG-1 stimulated TNF-α production. Taken together, we conclude that PSG-1 induces TNF-α secretion through TLR4/ROS/PI3K/Akt/MAPKs/NF-κB pathways during macrophage activation. Our findings provide a molecular basis for the potential of PSG-1 as a novel immunomodulatory agent.     

灵芝多糖对小鼠巨噬细胞白介素1α和肿瘤坏死因子αmRNA表达的影响

采用 RT- PCR和凝胶图像吸光度分析系统检测灵芝多糖 (GLB7)对小鼠腹腔巨噬细胞白介素 1α(IL- 1α) ,肿瘤坏死因子α(TNF-α) m RNA表达的影响 .加入不同浓度的 GLB7(5,1 0 ,2 0 ,40 mg·L-1) ,培养 3及 3.5h后观察 GLB7浓度与 IL- 1 α和 TNF- α m RNA表达之间的量效关系 .发现GLB7能浓度依赖性诱发 IL- 1 α和 TNF- α m RNA的表达 ,而对照组未见有 IL- 1 α和 TNF- α m RNA的表达 ;3及 3.5h培养上清中 IL- 1 α和 TNF- α活性与对照组比较亦有明显升高 ,并具有一定的浓度依赖关系 .说明 GLB7免疫增强和抗肿瘤作用的基础与其在转录水平诱发 IL- 1 α和 TNF- α m RNA的表达有关 .     

灵芝多糖对小鼠巨噬细胞白介素1α和肿瘤坏死因子α mRNA表达的影响

采用RT-PCR和凝胶图像吸光度分析系统检测灵芝多糖（GLB₇）对小鼠腹腔巨噬细胞白介素1α(IL-1α),肿瘤坏死因子α(TNF-α) mRNA表达的影响. 加入不同浓度的GLB₇（5,10,20,40 mg·L^-1）, 培养3及3.5 h后观察GLB₇浓度与IL-1α和TNF-α mRNA表达之间的量效关系. 发现GLB₇能浓度依赖性诱发IL-1α和TNF-α mRNA的表达,而对照组未见有IL-1α和TNF- α mRNA的表达;3及3.5 h培养上清中IL-1α和TNF-α活性与对照组比较亦有明显升高,并具有一定的浓度依赖关系. 说明GLB7免疫增强和抗肿瘤作用的基础与其在转录水平诱发IL-1α和TNF-α mRNA的表达有关.     

DATS通过NF-κB抑制LPS诱导小鼠肺泡巨噬细胞促炎细胞因子表达

目的探讨二烯丙基三硫(DATS)抑制脂多糖(LPS)诱导小鼠肺泡巨噬细胞肿瘤坏死因子-α(TNF-α)及白介素-1β表达的信号转导机制。方法体外培养MH-S细胞,用DATS和(或)LPS进行干预。反转录PCR检测细胞中TNF-α、IL-1β mRNA表达,电泳迁移率改变分析(EMSA)检测细胞核因子-κB(NF-κB)活性,Western blot检测细胞磷酸化(p-IκB)及非磷酸化IκB的表达。结果LPS刺激MH-S细胞可导致TNF-α、IL-1β mRNA、p-IκB表达增加及NF-κB活性升高。用DATS(0.1、0.5、2.5、5.0mg.L-1)预处理细胞30min后再给予LPS刺激,可使TNF-α、IL-1β mRNA表达降低,并呈剂量依赖性;升高的NF-κB活性及p-IκB表达均显不同程度的抑制。单独DATS对TNF-α、IL-1β mRNA表达及NF-κB活性无影响。结论DATS可通过抑制IκB磷酸化及NF-κB活化,进而下调LPS诱导小鼠肺泡巨噬细胞TNF-α、IL-1β mRNA表达。     

猪苓多糖通过Toll样受体4对小鼠腹腔巨噬细胞的活化作用

目的观察猪苓多糖(PPS)对小鼠腹腔巨噬细胞的活化作用,并研究Toll样受体4(TLR4)在其中的作用。方法用PPS12.5,25,50及100mg·L-1刺激C3H/HeN小鼠和TLR4缺陷的C3H/HeJ小鼠脾细胞72h或腹腔巨噬细胞24h,以[3H]TdR掺入法检测脾细胞增殖反应;以Griess法测定腹腔巨噬细胞培养上清一氧化氮(NO)水平,ELISA检测白细胞介素1β(IL-1β)和肿瘤坏死因子α(TNF-α)含量;以溴化氰活化多糖的方法制备荧光胺(Flu)标记的PPS(Flu-PPS)及Flu-葡聚糖,应用流式细胞仪及激光共聚焦显微镜检测Flu-PPS与小鼠腹腔巨噬细胞的结合。结果 PPS可明显促进C3H/HeN小鼠脾细胞增殖并诱导腹腔巨噬细胞分泌NO,IL-1β和TNF-α,且具有浓度依赖性(r=0.999,P<0.01)。用抗TLR4单抗20mg·L-1与C3H/HeN小鼠腹腔巨噬细胞预孵育1h后加入PPS50mg·L-1,与单独PPS50mg·L-1孵育比较,巨噬细胞培养上清IL-1β和TNF-α浓度分别由(0.38±0.06)μg·L-1和(0.29±0.05)μg·L-1降至(0.18±0.01)μg·L-1和(0.12±0.02)μg·L-1,降幅达52.6%和58.6%(P<0.01)。PPS对C3H/HeN小鼠的脾细胞增殖、腹腔巨噬细胞产生IL-1β和TNF-α的作用明显强于C3H/HeJ小鼠:PPS12.5～100mg·L-1组脾细胞增殖分别增加了2.4,1.7,1.5和22.2倍,IL-1β增加了0.9,1.3,1.2和1.1倍,TNF-α增加了1.3,0.9,0.6和0.6倍,差异均有统计学意义(P<0.01)。流式细胞仪及激光共聚焦显微镜分析结果表明,Flu-PPS1mg·L-1与小鼠腹腔巨噬细胞结合的荧光强度显著高于Flu-葡聚糖,增加Flu-PPS浓度至5mg·L-1,荧光强度并未相应增强,表明Flu-PPS与小鼠腹腔巨噬细胞的结合具有饱和性;未标记的PPS100mg·L-1及抗TLR4单抗200mg·L-1可明显阻断Flu-PPS与巨噬细胞的结合。结论 PPS可能通过TLR4活化小鼠腹腔巨噬细胞。     

吲哚硒对免疫性肝损伤小鼠一氧化氮和炎性细胞因子的影响

目的　研究吲哚硒(IND -Se)对免疫性肝损伤小鼠的影响。方法　建立BCG+LPS诱导免疫性肝损伤小鼠模型,分光光度法检测血清中丙氨酸氨基转移酶(alanineaminotrans ferase,ALT)、天门冬氨酸氨基转移酶(aspartateaminotrans ferase,AST)活性,Griess法检测血清中NO水平,放免法检测TNF α含量, MTT法检测腹腔巨噬细胞培养上清液中IL 1水平。结果　IND Se(5、10、20mg·kg-1 )灌胃给药可明显降低血清中ALT、AST活性,减轻肝组织坏死范围及程度,减少炎细胞浸润;进一步研究发现IND Se可降低血清中升高的NO和TNF -α水平,抑制免疫性肝损伤小鼠腹腔巨噬细胞TNF -α和IL-1的产生。结论　IND Se可减轻免疫性肝损伤小鼠的病理改变,其机制与降低血清NO和TNF- α水和抑制腹腔巨噬细胞炎性细胞因子的产生有关。     

Polysaccharide from Ganoderma atrum induces tumor necrosis factor-α secretion via phosphoinositide 3-kinase/Akt,mitogen-activated protein kinase and nuclear factor-κB signaling pathways in RAW264.7 cells

The aim of this study was to investigate the signaling pathways involved in the macrophage activation by Ganoderma atrum polysaccharide (PSG-1) and elucidate the molecular mechanism of PSG-1-induced signal transduction in the regulation of tumor necrosis factor (TNF)-α secretion. Our results illustrated that the mitogen-activated protein kinase (MAPK) pathways were simultaneously activated and involved in PSG-1-induced TNF-α secretion in RAW264.7 cells. Moreover, our results also demonstrated that the phosphoinositide 3-kinase (PI3K)/Akt pathway was stimulated and played an important role in the PSG-1 induced TNF-α secretion. Additionally, the present study showed that nuclear factor (NF)-κB activation by PSG-1 was triggered by PI3K/Akt/MAPK pathway and NF-κB participated in PSG-1 stimulated TNF-α production. In conclusion, we have elucidated the mechanism of PSG-1-mediated immunomodulatory activities, and provide a theoretical basis for the potential of PSG-1 as a novel immunomodulating agent.     

Rhynchophylline prevents cardiac dysfunction and improves survival in lipopolysaccharide-challenged mice via suppressing macrophage I-κBα phosphorylation

Myocardial dysfunction is a common complication during sepsis and significantly contributes to the mortality of patients with septic shock. However, none of the available therapeutic strategies proven to be effective in patients with severe sepsis are designed specifically to target myocardial dysfunction. The purpose of the present study is to investigate the effect of rhynchophylline (Rhy) on LPS-induced myocardial dysfunction in mice. We found that pretreatment with Rhy significantly improved cardiac systolic dysfunction, increased stroke volume and cardiac output in mice challenged with LPS. LPS induced cardiac inhibitor-κBα (I-κBα) phosphorylation, tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) mRNA expression, and in turn increased cardiac TNF-α and IL-1β protein production, all of which were attenuated by pretreatment with Rhy. Immunohistochemistry revealed that TNF-α was found in infiltrated macrophages (F4/80(+)) and myocardium, and Rhy reduced TNF-α immunostaining in cardiac infiltrated macrophages in LPS-challenged mice. Furthermore, Rhy inhibited LPS-induced I-κBα phosphorylation and TNF-α production in cultured mouse peritoneal macrophages, but not in neonatal mouse cardiomyocytes. Pretreatment with Rhy significantly decreased the mortality of LPS-challenged mice. These results indicate that Rhy reduces cardiac dysfunction and improves survival via suppression of macrophage I-κBα phosphorylation in LPS-challenged mice, and suggest that Rhy may be a potential agent for the treatment of septic cardiac dysfunction.     

Salvianic acid A inhibits induction of inflammatory mediators by blocking Nuclear Factor-kB activation in macrophages

Objective To investigate the anti-inflammation effect and possible mechanism of Salvianic acid A (SAA) in mouse peritoneal macrophages. Methods Peritoneal macrophages were obtained from BALB/c mice. LPS induced nitric oxide (NO), tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in supernatant, protein expression of inducible nitric oxide synthase (iNOS), matrix metalloproteinase-9 (MMP-9) and activation of nuclear factor-kappa B (NF-kB) in the extract were measured. Results SAA strongly inhibited the excessive production of NO, TNF-α and IL-6 in LPS-induced peritoneal macrophages in a concentration-dependent manner and blocked the expression of iNOS and MMP-9. Treatment with LPS alone increased the translocation of NF-kB (1065) from cytosol to the nucleus, but the SAA inhibited the translocation of NF-kB (p65). Conclusions The results showed that SAA had strong anti-inflammatory effects in LPS-stimulated peritoneal macrophages. The important mechanism is due to its inhibition of NF-kB activation.