灵芝多糖对小鼠腹腔巨噬细胞蛋白激酶C活性的影响

目的　观察灵芝多糖在体外对小鼠腹腔巨噬细胞(MΦ)蛋白激酶C(PKC)活性是否有影响。方法　采用反相离子对高效液相色谱法测定PKC活力。结果　灵芝多糖GLB7( 40mg·L-1)能引起小鼠腹腔MΦ中PKC活性明显升高 ,30min达峰值 ,2h恢复到基础水平 ;GLB7还可引起MΦ中PKC发生质膜转位 ,并拮抗Staurosporine对MΦ中PKC的抑制作用。结论　灵芝多糖的免疫增强作用与其增强PKC活性有关     

灵芝多糖对小鼠腹腔巨噬细胞cGMP含量的影响

目的 :研究灵芝多糖 (GLB7)对巨噬细胞 (MΦ)内cGMP含量的影响 ,为其免疫增强作用机制提供依据。方法 :采用竞争性蛋白结合分析法测定培养的小鼠腹腔MΦ中cGMP含量。结果 :GLB7 能剂量依赖性引起小鼠腹腔中cGMP浓度快速升高 ,10min达峰值 ,之后缓慢下降 ,至30min时基本恢复至原来水平。结论 :GLB7 引起小鼠MΦ内cGMP浓度快速升高 ,可能与其增强小鼠免疫功能有关     

灵芝多糖对小鼠T细胞蛋白激酶A和蛋激酶C活性的影响

:目的 :观察灵芝多糖体外对小鼠T细胞蛋白激酶A (PKA)和蛋白激酶C(PKC)活性是否有影响。方法 :采用反相离子对高效液相色谱法测定PKA和PKC活力。结果 :灵芝多糖GLB7 能引起T细胞中PKA和PKC活性明显增强并具有剂量依赖性 ,达峰时间分别为5min和20min ,于20min及1h分别恢复到基础水平 ;GLB7 还可引起T细胞中PKC发生质膜转位 ,并拮抗Stau rosporine对T细胞中PKC的抑制作用。结果 :灵芝多糖的免疫增强和抗肿瘤作用与其增强PKA和PKC活性有关     

螺旋藻多糖对小鼠腹腔巨噬细胞游离Ca~(2+)浓度的影响

目的 :研究探讨螺旋藻多糖 (SPP)的免疫调节机制。方法 :采用荧光分光光度法 ,测定SPP对小鼠腹腔巨噬细胞 (MΦ)游离Ca2 +浓度 ([Ca2 +]i)的影响。结果 :SPP能剂量依赖性地引起小鼠腹腔MΦ[Ca2 +]i 明显升高 ,[Ca2 +]i 的升高是由胞外钙内流和胞内钙释放共同作用的结果。结论 :SPP所产生的免疫调节作用与其能够使巨噬细胞[Ca2 +]i 升高有关。     

芪附汤对小鼠巨噬细胞活性及介质的影响

目的:芪附汤对小鼠巨噬细胞活性及介质的影响.方法:制备阳虚小鼠模型.用芪附汤对小鼠进行对照性喂养14d后,测定其腹腔巨噬细胞的活性及分泌的介质.结果:芪附汤对阳虚组和正常对照组影响相比:腹腔MΦ活性下降(P<0.01),分泌IL-1和TNF的能力下降(P<0.05),分泌NO增加与正常对照组相比有差异(P<0.05).对阳虚组精氨酸酶的活性和SOD的含量与正常对照组相比明显降低(P<0.01).结论:芪附汤可增强小鼠、特别是阳虚小鼠腹腔MΦ的活性,分泌大量的NO、IL-1、TNF发挥效应.     

毛木耳多糖对小鼠腹腔巨噬细胞胞浆游离Ca^2＋浓度的影响

为探讨毛木耳多糖（APSP－A）对免疫细胞信号转导的影响，采用荧光分光光度法，测定毛木耳多糖（APSP－A）对小鼠腹腔巨噬细胞（Mφ）胞浆游离Ca2 浓度（[Ca2 ]i）的影响。结果表明：APSP-A能剂量依赖性地引起小鼠腹腔Mφ[Ca2 ]i明显升高，[Ca2 ]i的升高是由胞外钙内流和胞内钙释放共同作用的结果;与Ca2 通道有关，而与细胞膜电位无关。     

毛木耳多糖的抗氧化作用

目的：观察毛木耳多糖的抗氧化作用。方法：毛木耳多糖按剂量小鼠腹腔注射,ｑｄ,连续３５ｄ。以分光光度法测定小鼠全血中ＬＰＯ含量,按邻苯三酚自氧法测定小鼠全血ＳＯＤ的活力。结果：毛木耳多糖能剂量依赖性降低小鼠血清中ＬＰＯ含量,并可剂量依赖性地提高小鼠全血ＳＯＤ活力。结论：毛木耳多糖表现出较好的抗氧化作用,提示其具有一定的抗衰老作用。     

防芪汤及其组方对免疫抑制小鼠免疫功能的影响

目的:探讨防芪汤及其组方对免疫抑制小鼠免疫功能的影响。方法:建立受氢化可的松(HC)免疫抑制动物模型,应用防芪汤及其组方对免疫抑制小鼠进行治疗,观察其对免疫抑制小鼠免疫功能的影响。结果:防芪汤可明显增强免疫抑制小鼠腹腔巨噬细胞(MΦ)的活性和红细胞C_3b受体(RBC—C_3bR)花环形成率、脾细胞产生抗体的能力及白介素-2(IL—2)的活性,增强红细胞免疫复合物(RBC-IC)花环形成率和脾细胞增埴反应。黄芪配伍水煎液能促进免疫抑制小鼠的腹腔MΦ的活性和IL—2的活性,增强脾细胞产生抗体的能力,但作用不及防芪汤;防已配伍水煎液能促进免疫抑制小鼠脾细胞产生抗体,作用与黄芪配伍水煎液相近。结论:防芪汤能增强受HC免疫抑制小鼠的各项免疫功能。     

北沙参粗多糖的提取及对阴虚小鼠的免疫调节作用

目的研究北沙参粗多糖(GLP)的滋阴和免疫调节作用。方法提取GLP,制备阴虚小鼠模型,观察对小鼠体重变化、脾脏抗体生成细胞(AFC)、迟发型超敏反应(DTH)和腹腔巨噬细胞吞噬功能的影响。结果GLP可使阴虚小鼠体重明显增加;亦能显著增加阴虚小鼠脾脏AFC的数量,增强DTH反应,而对腹腔MΦ的吞噬百分率和吞噬指数无明显影响。结论GLP具有滋阴补虚作用,可增强体液免疫和细胞免疫功能。     

芪附汤对小鼠巨噬细胞活性及介质的影响

目的芪附汤对小鼠巨噬细胞活性及介质的影响.方法制备阳虚小鼠模型.用芪附汤对小鼠进行对照性喂养14d后,测定其腹腔巨噬细胞的活性及分泌的介质.结果芪附汤对阳虚组和正常对照组影响相比腹腔MΦ活性下降(P<0.01),分泌IL-1和TNF的能力下降(P<0.05),分泌NO增加与正常对照组相比有差异(P<0.05).对阳虚组精氨酸酶的活性和SOD的含量与正常对照组相比明显降低(P<0.01).结论芪附汤可增强小鼠、特别是阳虚小鼠腹腔MΦ的活性,分泌大量的NO、IL-1、TNF发挥效应.     

甲壳胺体外对小鼠巨噬细胞PKC和DAG的作用研究

目的 观察甲壳胺体外对小鼠腹腔巨噬细胞蛋白激酶C(PKC)活性和二酰基甘油(DAG)合成的影响。方法分别采用反相离子对高效液相色谱法与放射免疫测定技术测定PKC活力和DAG含量。结果 ①甲壳胺剂量依赖性引起巨噬细胞中PKC活性明显增强并使之发生质膜转位,达峰时间为25min,1.5 h恢复到基础水平;②甲壳胺促进巨噬细胞中DAG的合成,达峰时间为30 s,3 min恢复到基础水平。结论 甲壳胺的免疫增强作用与其增强巨噬细胞中PKC活性和DAG合成有关,DAG/PKC信号途径参与了其对巨噬细胞的活化过程。     

Immunological and anti-inflammatory effects of clarithromycin: inhibition of interleukin 1 production of murine peritoneal macrophages

The immunological and anti-inflammatory effects of clarithromycin (CAM), a new oral macrolide antibiotic, were examined in in vitro models such as lymphocyte transformation (LTF) of murine spleen cells, interleukin 1 (IL-1) production of murine peritoneal macrophages and IL-1-induced proliferation of C3H/HeJ mice thymocytes; the results were compared with those achieved by erythromycin (EM). CAM suppressed these responses much more than EM. Murine peritoneal macrophages precultured with CAM showed diminished IL-1 production, but macrophages precultured with EM did not, indicating that CAM has suppressive effects on the early phase of IL-1 production of murine peritoneal macrophages. Suppressive effects of CAM on IL-1 production by macrophages and proliferation of lymphocytes were independent of prostaglandin biosynthesis, since this drug had no effect on cyclooxygenase activity. Additional immunosuppressive and anti-inflammatory activities of CAM may explain its superior clinical effect.     

Modulation of murine macrophage function by methamphetamine

The abuse of methamphetamine (MA) is an increasingly growing problem globally and produces serious side effects. In the present study, the immunomodulating effects of MA were examined on murine peritoneal macrophages after MA (5 mg/kg body weight) was administered daily orally for 2 wk. When purified macrophages were stimulated with lipopolysaccharide (LPS), the tumoricidal activity induced by LPS was significantly suppressed by MA. MA also inhibited poly I:C-induced antiviral activity in macrophages and decreased the number of peritoneal macrophages. FACS analysis showed that the expression of CD14 was markedly decreased by MA in LPS-stimulated macrophages. The production of nitric oxide (NO) and tumor necrosis factor (TNF)-alpha: which are known to be major effector molecules in macrophage-mediated cytotoxicity, was decreased by MA. MA produced a significant effect on phagocytosis and interleukin-1 (IL-1) and IL-6 at 14 d. In addition, the level of hydrogen peroxide (H2O2) was not altered by MA. Taken together, these data indicate that MA has a differential immunomodulating effect on macrophage secretory and cellular activities.     

Stimulation of various functions in murine peritoneal macrophages by high mannuronic acid-containing alginate (HMA) exposure in vivo

High mannuronic acid-containing alginate (HMA) was tested to affect murine peritoneal macrophages. In the present study, we measured various functions of murine peritoneal macrophages that were isolated 20 h after intraperitoneal injection with HMA (25 and 100 mg/kg). HMA increased the number of peritoneal macrophages and phagocytosis. Macrophages from HMA-treated mice significantly inhibited growth of tumor cells compared to macrophages from control mice. In addition, supernatants from macrophages of HMA-treated mice contained nitric oxide (NO), hydrogen peroxide (H2O2) and TNF-alpha. The increased production of these cytotoxic molecules induced by HMA is consistent with tumoricidal activity of activated macrophages. Furthermore, HMA-induced tumoricidal activity was partially abrogated by anti-TNF-alpha, inhibitors of NO and the scavenger of reactive oxygen. Thus, the tumoricidal activity induced by HMA appeared to be mediated by the production of TNF-alpha, NO and H2O2. Taken together, these results suggest that HMA has the immunostimulating effect on macrophages after in vivo exposure of it.     

银杏内酯B对血小板活化因子引起的巨噬细胞趋化及细胞骨架改变的影响

目的考察银杏内酯B(ginkgolide B，BN52021)对血小板活化因子(PAF)引起的小鼠腹腔巨噬细胞趋化反应和丝状肌动蛋白(F-actin)聚合作用的影响。方法Boyden小室法检测化合物对巨噬细胞趋化的影响；流式细胞术检测特异性标记的巨噬细胞中F-actin的变化。结果PAF可显著刺激小鼠腹腔巨噬细胞产生趋化效应，PAF受体拮抗剂BN52021(0.01 nmol·L^-1～0.1 μmol·L^-1)可明显抑制该作用。此外，在含钙缓冲液中，BN52021可显著抑制PAF引起的丝状肌动蛋白聚合。结论BN52021可能通过抑制丝状肌动蛋白的聚合作用，从而抑制PAF引起的巨噬细胞趋化，并且这种作用是钙依赖性的。表明BN52021的抗炎作用途径之一是抑制PAF诱导的趋化作用。     

赤芍801对小鼠巨噬细胞COX-1和COX-2活性、mRNA及蛋白表达的影响

目的研究赤芍801(propyl gallate,PrG)对环氧酶(cy-c looxygenase,COX)活性、mRNA及蛋白表达的影响。方法采用基于小鼠腹腔巨噬细胞的COX-1和COX-2体外筛选模型,用钙离子导入剂(calc ium ionophore A23187)短时刺激小鼠腹腔巨噬细胞,测定培养上清液中的6酮--前列腺素F1α(6-keto-PGFlα)的量反映COX-1活性;用脂多糖(lipopolysac-charide,LPS)长时间刺激细胞,测定培养上清液中的前列腺素E2(PGE2)的量反映COX-2活性。半定量RT-PCR法检测PrG对LPS刺激的Raw小鼠巨噬细胞COX-1、COX-2 mRNA表达的影响。W estern b lot法检测PrG对LPS刺激的Raw小鼠巨噬细胞COX-1、COX-2蛋白表达的影响。结果PrG体外10,5μmol.L-1浓度下不影响6-keto-PGFαl生成(P>0.05),而在1,0.5,0.1,0.05μmol.L-1浓度下则诱导6-keto-PGF1α生成(P<0.01),并呈较好的剂量依赖性。PrG体外1,10μmol.L-1可抑制PGE2生成(P<0.05)。PrG不同浓度对LPS刺激的Raw小鼠巨噬细胞COX-1、COX-2 mR-NA表达无明显影响。PrG(100,10,1μmol.L-1)浓度下可抑制COX-2蛋白表达,但对COX-1蛋白表达无影响。结论在0.05~10μmol.L-1浓度范围内,PrG低浓度时促进6-keto-PGF1α生成,高浓度时抑制PGE2生成,提示其高浓度时抑制COX-2,低浓度时激活COX-1;不同浓度PrG对COX-1、COX-2 mRNA表达均无明显影响。PrG(1~100μmol.L-1)浓度下可抑制COX-2蛋白表达,但对COX-1蛋白表达无影响。     

In vitro investigation on the effect of a plant preparation with antiviral activity on the functions of mice phagocyte cells

A polyphenol extract from the aerial roots of the medicinal plant Geranium sanguineum L. (PC) inhibited the reproduction of influenza viruses type A and B in vitro and in ovo and protected mice from mortality in the experimental influenza infection. The in vivo protective effect was connected with multiple biological activities of the preparation. The present paper focuses on the in vitro effects of the polyphenol extract on the functions of peritoneal and alveolar macrophages and blood polymorphonuclear leucocytes (PMNs), isolated from healthy ICR mice. It was found that PC in doses of 12.5 and 25 microg ml(-1) stimulated the phagocytic activity of peritoneal macrophages and blood PMNs. PC in the same doses did not significantly affect the phagocytic activity of alveolar macrophages, the migration of alveolar and peritoneal macrophages or the adherent activity of PMNs. Used in concentrations of 3.1-25.0 microg ml(-1), PC suppressed spontaneous NO production from peritoneal macrophages, while inducible NO production, provoked by LPS-, Ifn-gamma and LPS + Ifn-gamma inductions was not affected. The cell-toxic concentration of 100 microg ml(-1) increased spontaneous and LPS-inducible NO production. The experimental results demonstrated a stimulating effect of PC on the phagocytic activity of murine PMNs and peritoneal macrophages as well as a beneficial effect of the preparation on spontaneous NO production.     

灵芝多糖对小鼠腹腔巨噬细胞活性氧自由基的影响

为进一步探讨灵芝多糖 ( GLP)免疫调节作用机理 ,采用激光扫描共聚焦显微镜技术 ,动态监测灵芝多糖均一体 GLB7对小鼠腹腔巨噬细胞 ( M )活性氧自由基含量的影响 .以 GLB7( 2 0 mg· L-1)刺激 M ,发现探针 DCHF- DA的荧光强度明显下降 ,与静息水平相比 ,平均下降 ( 36± 6) % ( n=6,P<0 .0 5) ;同时还能拮抗 PMA引起的 M 呼吸爆发 ,荧光强度下降幅度为 ( 1 9± 4) % ( n=6,P<0 .0 5) .结果证明 :GLB7能减少 M 内活性氧自由基的生成 ,具有清除活性氧的作用 ,这也是 GLB7产生免疫增强和抗衰老作用的重要途径 .