Lys-2-rhTNFα及~(125)Ser-rhIL-2促进小鼠腹腔巨噬细胞的肿瘤杀伤活性的研究

研究了 L ys- 2 - rh TNFα(1)及 1 2 5 Ser- rh IL- 2 (2 )对小鼠腹腔巨噬细胞杀伤肿瘤的促进作用。2单用能激活小鼠巨噬细胞对小鼠肥大细胞瘤 P815的细胞毒性 ,1单独无此作用 ,但与 2联合使用能促进 2对巨噬细胞的激活作用。腹腔非粘附细胞的存在是 2或 2联合 1激活巨噬细胞活性的重要条件     

槲寄生凝集素抑制人大肠癌HCT116细胞环氧化酶-2表达的研究

目的：探讨槲寄生凝集素对人大肠癌HCT116细胞环氧化酶（COX-2）表达的影响。方法：采用RT-PCR、Western blotting和ELISA等方法检测不同浓度（0.1～1mg/L）槲寄生凝集素对细胞COX-2及其产物前列腺素E2（PGE2）表达的影响。结果：槲寄生凝集素可抑制HCT116细胞的COX-2mRNA、蛋白表达及PGE2的水平,其抑制COX-2蛋白的表达与PGE2的水平相一致,只在大剂量（1mg/L）时对COX-1mRNA有抑制作用。结论：槲寄生凝集素具有抑制HCT116细胞的COX-2基因转录、蛋白表达和功能活性等作用,在一定浓度（0.1～1mg/L）和时间范围（3～24h）内,表现出时效与量效关系。     

黑灵芝多糖对体外培养的小鼠腹腔巨噬细胞功能的影响

目的研究黑灵芝多糖对小鼠腹腔巨噬细胞功能的影响。方法用不同浓度的黑灵芝多糖作用于正常的和LPS活化的腹腔巨噬细胞;测定巨噬细胞代谢MTT活力;Griess法测定NO的产生;ELISA法检测巨噬细胞培养上清中TNF-α、IL-1β的分泌水平。结果黑灵芝多糖对细胞代谢MTT活力有增强作用;在20～160mg·L-1范围内,多糖呈剂量依赖性地促进正常的巨噬细胞分泌NO、TNF-α、IL-1β,增强免疫;而巨噬细胞经LPS激活后,与LPS组比较,多糖不同程度地抑制NO、TNF-α、IL-1β的过量分泌。结论黑灵芝多糖能有效地增强小鼠腹腔巨噬细胞的免疫,可改善LPS对小鼠腹腔巨噬细胞的诱导作用。     

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。     

Lys-2-rhTNFα及125Ser-rhIL-2促进小鼠腹腔

研究了Lys-2-rhTNFα(1)及 125Ser-rhIL-2(2)对小鼠腹腔巨噬细胞杀伤肿瘤的促进作用.2单用能激活小鼠巨噬细胞对小鼠肥大细胞瘤P815的细胞毒性,1单独无此作用,但与2联合使用能促进2对巨噬细胞的激活作用.腹腔非粘附细胞的存在是2或2联合1激活巨噬细胞活性的重要条件.     

多抗甲素诱导大鼠腹腔巨噬细胞对癌细胞杀伤增强机制的研究

目的：探讨多抗甲素增强大鼠腹腔巨噬细胞对癌细胞杀伤作用的机制。方法：大鼠腹腔内分别注射0．5，1．0，1．5mg的多抗甲素。2d后处死大鼠分离巨噬细胞，计数并测定乳酸脱氢酶(LDH)和酸性磷酸酶(ACP)的活性、巨噬细胞吞噬活力、一氧化氮(NO)的分泌及巨噬细胞对人K562细胞的细胞杀伤性。结果：大鼠腹腔巨噬细胞数量、LDH和ACP的活性、吞噬活力、NO的分泌以及细胞的杀伤性随着多抗甲素剂量的增加而增加。结论：多抗甲素通过3种方式增加腹腔巨噬细胞杀伤癌细胞作用，(1)诱导大量的巨噬细胞进入腹腔；(2)增强这些细胞的吞噬、清除能力；(3)增强巨噬细胞的非接触性细胞杀伤性。     

槲寄生通过抑制NF-κB活性来诱导人大肠癌HCT116细胞凋亡

目的：探讨槲寄生诱导人大肠癌HCT116细胞凋亡所涉及的信号转导途径。方法：采用MTT法测定细胞的增殖活力,琼脂糖凝胶电泳观察细胞调亡,流式细胞仪进行细胞周期分析,Western blotting和DIG-EMSA研究细胞调亡途径。结果：槲寄生对HCT116细胞有明显的生长抑制作用,琼脂糖凝胶电泳检测到典型的DNA梯状条带;槲寄生可通过抑制IκBα蛋白磷酸化进而抑制NF-κB;槲寄生可增强羟基喜树碱对HCT116细胞诱导的凋亡作用。结论：槲寄生对HCT116细胞有明显的生长抑制和诱导调亡作用,并且通过抑制NF-κB活性来诱导人大肠癌HCT116细胞凋亡。     

黄芪皂苷Ⅳ对小鼠T、B淋巴细胞增殖和腹腔巨噬细胞功能的影响

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L9292细胞杀伤法测定。结果：1）ASI50－200mg/kg ig7天能够促进T淋巴细胞增殖和抗体生成,而ASI50－100mg/kg 能够促进B淋巴细胞增殖,但是200mg/kg对B淋巴细胞增殖无影响;（2）ASI体外仅在200nmol/L对T,B淋巴细胞有促进作用;（3）ASI 1nmol/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内分泌IL－1和TNF－α。     

海带多糖对小鼠腹腔巨噬细胞的激活作用

本文研究了海带多糖对Ｃ５７ＢＬ／６小鼠腹腔巨噬细胞的激活作用,结果表明,腹腔注射海带多糖能够明显激活小鼠腹腔巨噬细胞,增强其细胞溶解作用。海带多糖激活小鼠腹腔巨噬细胞,在有ＬＰＳ存在的条件下,能在体外分泌肿瘤坏死因子。     

Lys—2—rhTNFα及^125Ser—rhIL—2促进小鼠腹腔巨噬细胞的肿瘤杀伤活性的研究

研究了Lys-2-rhTNFα(1)及^125Ser-rhIL-2(2)对小鼠腹腔巨噬细胞杀伤肿瘤的促进作用。2单用能激小鼠巨噬细胞对小鼠肥大细胞瘤P815的细胞毒性，1单独无此作用，但与2联合使用能促进2对巨噬细胞的激作用，腹腔非粘附细胞的存在是2或2联合1激活巨噬细胞活性的重要条件。     

Liposomal Induction of a Heat-stable Macrophage Priming Factor to Induce Nitric Oxide in Response to LPS

Purpose. The effects of liposomes on nitric oxide (NO) production from mouse peritoneal macrophages following intraperitoneal injection of liposomes were investigated. Methods. Mouse peritoneal macrophages were collected following intraperitoneal injection of liposomes and cultured with and without lipopolysaccharide (LPS). Peritoneal washing fluid was also collected from the mice injected with liposomes. NO production was evaluated by measuring the concentration of nitrite in the macrophage culture supernatant by Griess reagent. Results. NO production stimulated by LPS was observed in peritoneal macrophages obtained from the liposome-treated mice, but liposomes did not activate macrophages directly to induce NO in response to LPS. NO production was higher in the liposomes composed of phospha-tidylcholine than that of negatively charged liposomes composed of phosphatidylserine. Peritoneal washing fluid obtained from mice injected with liposomes has a capacity to induce NO production in the macrophages from naive mice. This capacity was not diminished by heat-treatment at 100°C for 5 min. Conclusions. Peritoneal macrophages were activated to produce NO in response to LPS following intraperitoneal injection of liposomes. They did not activate macrophages directly, and the induction of heat-stable macrophage priming factor, but not cytokines, is suggested.     

Activation of macrophage function by intraperitoneal administration of the streptococcal antitumor agent OK-432

Motility, adhesiveness, IL1 production, and inhibition of tumor cell growth in vitro were examined in murine peritoneal macrophages obtained after intraperitoneal injection of a streptococcal preparation OK-432, heat-inactivated OK-432 (HI-OK-432), and thioglycollate medium (TG). By varying the interval between intraperitoneal injection of OK-432 and the harvest of peritoneal macrophages, it was found that OK-432 induced a time-dependent multi-step alteration of these properties: step I increased motility on day 1: step II increased adhesiveness on day 2; and step II increased inhibition of tumor cell growth and IL1 production. During step III, the peritoneal macrophage population, including Ia-bearing cells, increased dramatically in the peritoneal cavities of OK-432-treated mice. In contrast, injection of either HI-OK-432 or TG, which lack antitumor activity in vivo, initiated steps I and II, but not step III. The Ia-bearing macrophages induced by OK-432 showed high ability of IL1 production, but low growth inhibitory activity against tumor cells. Based on these results, OK-432 seems to be performing a dual function: eliciting a new population of macrophages to the site of injection (heat stable function), and inducing two different populations of antitumor macrophages and Ia-bearing macrophages (heat unstable function).     

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.     

Release of tumor necrosis factor (TNF) into mouse peritoneal fluids by OK-432, a streptococcal preparation

A cytotoxic factor was induced in peritoneal fluid by injection of OK-432 in mice which had been primed with OK-432. Two-step stimulation (priming and eliciting) was always necessary to induce the cytotoxic factor. OK-432-primed mice did not produce soluble cytotoxic factor spontaneously and no cytotoxic activity was detected in the mice treated by a single injection of OK-432 as an eliciting agent. This observation was also confirmed by in vitro experiments. Only when activated macrophages were incubated with OK-432 (or with lipopolysaccharide) was cytotoxin released into medium supernatant. High doses of OK-432 were required to prime mice for the production of cytotoxic factor, whereas a small amount was enough to elicit. The peritoneal cytotoxic factor obtained by OK-432 injection appears to be identical to tumor necrosis factor in the serum for the following reasons: The two factors are similar in mode of cytotoxic action. Both are produced from macrophages. They are similar in physicochemical characteristics. The cytotoxicity of the peritoneal cytotoxic factor was totally abolished by anti-TNF serum.     

灵芝多糖肽对小鼠腹腔巨噬细胞一氧化氮产生的影响

目的　研究灵芝多糖肽 (GLPP)对小鼠腹腔巨噬细胞一氧化氮产生的影响并探讨其作用机制。方法　以Griess法 ,观察GLPP对LPS诱导小鼠腹腔巨噬细胞一氧化氮(NO)产生的影响 ;以免疫组化法检测诱导型一氧化氮合成酶 (iNOS)的表达 ,观察GLPP对iNOS的影响。结果　GLPP(2 5～ 2 0 0mg·kg-1)灌胃给药 5d或体外给药 (3 12 5～ 2 0 0mg·L-1)均可促进巨噬细胞NO释放 ,但对LPS刺激NO的释放影响不大 ;GLPP(10 0mg·kg-1)灌胃给药 5d或体外给药 (10mg·L-1)均可使巨噬细胞iNOS含量增加。结论　GLPP可增加小鼠腹腔巨噬细胞NO产生 ,其机制可能与其促进巨噬细胞iNOS合成有关。     

糖萜素对小鼠腹腔巨噬细胞产生一氧化氮的影响

目的：探讨糖萜素对小鼠腹腔巨噬细胞产生NO的影响。方法：在饲料中添加不同剂量糖萜素喂养NIH小鼠，以基础饲料为对照，检测环磷酰胺免疫抑制组与非抑制组不同时间段小鼠腹腔巨噬细胞产生NO的含量。结果：含糖萜素饲料组于不同时间测得NO含量均较基础饲料组显著增高（P＜0．05）。免疫抑制状态下，含糖萜素饲料组NO含量也较基础饲料组高（P＜0．05）。体外糖萜素不能直接影响NO的生成。结论：糖萜素能显著提高小鼠体内巨噬细胞产生NO。体外则不能直接影响NO的生成。     

In vivo activation of functional properties in mouse peritoneal macrophages by Nocardia rubra cell wall skeleton

Exudative cells in the peritoneal cavity of mice, particularly polymorphonuclear leukocytes significantly increased 1 day after intraperitoneal injection of Nocardia rubra cell wall skeleton (N-CWS). Macrophages and lymphocytes significantly increased 4 to 7 days after injection. N-CWS also enhanced peritoneal macrophage functions such as phagocytosis of latex particles, production of superoxide anion, production and secretion of lysosomal enzymes such as beta-glucuronidase, lysozyme and acid phosphatase, phagocytosis and intracellular killing of bacteria, and in vitro chemotaxis. The phagocytic function of the reticuloendothelial system was also enhanced. These results indicate that macrophages were activated in vivo by N-CWS.     

Effects of mancozeb on the activities of murine peritoneal macrophagesin vitro andex vivo

Mancozeb (MCZ) is known to have detrimental effects on the reproductive system, but the toxicity of MCZ on immune responses has not been systematically investigated. We investigated the effects of MCZ exposure on the activities of murine peritoneal macrophages through evaluation of MCZ-induced alteration of nitric oxide (NO) production and tumor necrosis factor-alpha (TNF-alpha) synthesis. Macrophages were examined ex vivo from mice orally treated with various doses of MCZ for 5 consecutive days per week for 4 weeks (subacute exposure, 250, 1000, 1500 mg/kg/day) followed by culture for 2 (TNF-alpha) or 3 days (NO) in the presence of LPS plus IFN-gamma. Macrophages from naive mice were also cultured with various concentrations of MCZ (0.05, 0.25, 0.5, 1 and 2 microg/mIL in the presence of LPS plus IFN-gamma for 2 (TNF-alpha) or 3 days (NO) in vitro. NO production was decreased with the in vitro exposure to all concentrations of MCZ. However, the amount of NO production by peritoneal macrophages from MCZ-subacutely exposed mice was increased in comparision with that of control group. In vitro, MCZ suppressed TNF-alpha secretion with significant reduction at 2 microg/mL MCZ. Conversely, TNF-alpha release was enhanced ex vivo. This study provides the substantial evidence on MCZ-induced alternation in macrophage activity. In order to clearly understand the contrasting effect of MCZ on peritoneal macrophage activity, it is necessary to further investigate the influence of major metabolite of MCZ (ETU) exposure on the NO production and TNF-alpha synthesis.     

Potentiation of tumoricidal properties of murine macrophages by Nocardia rubra cell wall skeleton (N-CWS)

Intraperitoneal injection of squalene-treated cell wall skeleton of Nocardia rubra (N-CWS) caused increase in number of peritoneal exudate cells (PEC). Adherent macrophages obtained from N-CWS-treated PEC suppressed growth of methylcholanthrene-induced fibrosarcoma (Meth-A), when injected intradermally with the tumor cells into BALB/c mice. The macrophages showed strong cytotoxicity against Meth-A cells in vitro. When treated with 10 micrograms/ml of N-CWS in vitro, proteose peptone-induced macrophages acquired tumoricidal property but resident macrophages showed no cytotoxicity after the treatment. In the supernatant of spleen cells cultured for 72 hours in the presence of N-CWS (10 micrograms/ml), the presence of (a) factor(s) with macrophage activating effect was observed. This factor, shown to be identical to macrophage activating factor (MAF) in molecular weight, showed synergy with N-CWS in potentiating macrophage cytotoxicity against tumor cells.