槲寄生凝集素腹腔内给药对腹腔巨噬细胞功能的影响

目的：探讨小鼠腹腔内注射槲寄生凝集素对腹腔巨噬细胞功能的影响。方法：16只小鼠分为腹腔内给药组和空白组,分别取腹腔内巨噬细胞并计数,采用ELISA法测定TNF-α表达情况,Griess法测定NO表达,MTT法检测腹腔内巨噬细胞对肠癌HCT116细胞的杀伤作用。结果：腹腔内给槲寄生凝集素后可明显增加腹腔内巨噬细胞数,并增加TNF—α和NO的表达水平;激活的巨噬细胞对HCT116细胞的杀伤作用增强,其杀伤作用与TNF—α和NO表达时间一致。结论：腹腔内注射槲寄生凝集素可激活腹腔内巨噬细胞,激活的巨噬细胞对HCT116细胞有杀伤作用。     

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.     

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     

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。     

短小棒状杆菌免疫后可以看到粒细胞介导的巨噬细胞活化作用。

作者用短小棒状杆菌(CP)免疫小鼠,以阐明中性粒细胞在巨噬细胞被激活过程中的作用。实验采用贴壁巨噬细胞毒试验和细胞化学分析等方法,证明吞食CP的中性粒细胞有激活巨噬细胞的作用。试验用6～8周龄AB6F1小鼠,腹腔免疫CP700μg/0.1ml后5小时,将分离出的2～5×10~6个炎症中性粒细胞腹腔注射到正常小鼠。4天后收集受     

花色素苷诱导小鼠巨噬细胞一氧化氮生成及其机制

目的研究花色素苷对小鼠腹腔巨噬细胞一氧化氮(NO)合成的诱导作用及其机制。方法用CCK-8试剂检测花色素苷对小鼠脾细胞增殖的影响;硝酸盐还原酶法检测小鼠腹腔巨噬细胞NO含量;荧光法检测一氧化氮合酶(NOS)活性;RT-PCR检测iNOS mRNA的表达。结果花色素苷可促进小鼠脾细胞增殖,诱导小鼠腹腔巨噬细胞合成NO,提高NOS活性,其中矢车菊素-3-葡萄糖苷能够诱导iNOS mRNA的表达。结论花色素苷能够促进脾细胞的增殖,诱导小鼠腹腔巨噬细胞合成NO,使巨噬细胞激活,具有一定的免疫调节活性。     

雷公藤多甙诱导小鼠腹腔激活巨噬细胞凋亡的实验研究

目的体外观察雷公藤多甙对小鼠腹腔激活巨噬细胞凋亡的影响。方法分离小鼠腹腔巨噬细胞,以脂多糖(LPS)激活,与雷公藤多甙(TWP)进行共同孵育;分别采用夹心ELISA法检测巨噬细胞NF-кB的核内浓度及凋亡电泳试验显示巨噬细胞凋亡情况。结果 TWP在12.5～100μg/L范围内对LPS活化的巨噬细胞凋亡具有促进作用,呈剂量依赖关系。结论 TWP可以促进激活巨噬细胞的(Mф)凋亡。     

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。     

草乌甲素对小鼠腹腔巨噬细胞趋化功能的影响

目的研究草乌甲素（BLA）对小鼠腹腔巨噬细胞趋化功能的影响。方法小鼠腹腔巨噬细胞经分离后,制成细胞密度为1×10^6个·mL^-1的细胞悬液;而趋化液由4部分按不同比例组成,即细胞培养基RPMI-1640液,溶剂（0.01%DMSO）,BLA溶液（质量浓度分别为80、40、20、10、5μg·L^-1）和趋化因子（MCP-1或补体激活产物）。应用transwell 24孔趋化板检测腹腔巨噬细胞在不同趋化液中的趋化功能,以评价BLA对不同条件下细胞趋化作用的影响。结果 MCP-1和补体激活产物均可使腹腔巨噬细胞的趋化指数显著高于溶剂对照组（P〈0.01）,而该趋化指数可受到BLA溶液的抑制（P〈0.01）。但单用BLA（80、40、20、10、5μg·L^-1）对腹腔巨噬细胞的趋化指数与溶剂对照组相似,差异均无统计学意义（P〉0.05）。结论 BLA能抑制趋化因子对腹腔巨噬细胞的趋化作用,但对腹腔巨噬细胞无直接作用。     

何首乌对小鼠腹腔巨噬细胞功能的影响

为了解何首乌对小鼠腹腔巨噬细胞功能的影响,采用微量浊度法测定小鼠腹腔巨噬细胞介导的细胞毒活性;采用小鼠胸腺细胞检测法测定小鼠腹腔巨噬细胞白细胞介素-1的分泌活性;采用姬姆萨染色法检测小鼠腹腔巨噬细胞的吞噬功能。结果表明,各药物实验组小鼠腹腔巨噬细胞介导的细胞毒作用及泌白细胞介素-1的分泌活性明显增强(P<0.01);各药物实验组小鼠腹腔巨噬细胞的吞噬功能明显增强(P<0.01),提示何首乌能促进小鼠腹腔巨噬细胞功能。     

Macrophage activation reduces mobilization of arachidonic acid by guinea-pig and rat peritoneal macrophages in vitro

We have examined species differences in the mobilization of arachidonic acid and generation of prostacyclin in non-activated and activated peritoneal macrophages in vitro. Mobilization of 3H-arachidonic acid was reduced in rat activated macrophages compared with that in non-activated macrophages, but a similar difference was not observed in guinea-pig macrophages. In guinea-pig peritoneal macrophages, exposure to formyl-Methionyl-Leucyl-Phenylalanine (fMLP), platelet-activating factor (Paf), zymosan and A23187 increased the generation of prostacyclin. In contrast, in rat peritoneal macrophages, fMLP and Paf did not stimulate the mobilization of arachidonic acid or the generation of prostacyclin, whereas both zymosan and A23187 were effective stimuli. Pretreatment of either rats or guinea-pigs by intraperitoneal injection of C. Parvum reduced prostacyclin generation by peritoneal macrophages in vitro. We conclude that there may be species differences in receptor populations between guinea-pig and rat peritoneal macrophages. However, the reduction in eicosanoid generation induced by the inflammatory stimulus, C. Parvum is not species-dependent.     

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.     

Immunoactive peptides, FK 156 and FK 565. IV. Activation of mouse macrophages

We investigated the effects of the immunoactive peptides, FK 156 and FK 565, on functions of mouse macrophages. FK 156 and FK 565 given parenterally or orally to mice enhanced spreading of peritoneal macrophages, phagocytosis of latex particles and intracellular killing of bacteria by peritoneal macrophages. FK 156 and FK 565 also enhanced the production of superoxide anion and lysosomal enzyme activities of macrophages. The peptides also activated mouse spleen macrophages, and the kinetics of this activation differed from that of the peritoneal macrophages. In addition, both drugs directly enhanced the production of superoxide anion by mouse peritoneal macrophages treated in vitro and enhanced the functions of peritoneal macrophages of athymic nude mice. Both these phenomena suggest that direct activation might be one of the mechanisms of macrophage activation by the peptides.     

Human peritoneal macrophages: clinical models of inflammation and potential targets of antiinflammatory drugs

Human peritoneal macrophages have been obtained from patients with renal disease undergoing chronic peritoneal dialysis, patients with ascites and at laparoscopy. These macrophages in general have both morphological and enzymatic characteristics of activated macrophages, as judged by criteria derived from animal experiments. Human peritoneal macrophages produce a variety of eicosanoids, including leukotriene B4 and leukotriene C4. These cells are suitable for studies on in vitro and in vivo effects of drugs, and for investigation of changes in macrophage activity occurring in human diseases.     

Immunomodulating effect of killed, apathogenic Escherichia coli, strain Nissle 1917, on the macrophage system]

The influence of formaldehyde-killed Escherichia coli strain Nissle 1917 (SK 22) on macrophages of C57BL/6 mice was investigated in vitro. It has been shown that SK 22 activated macrophages derived from bone marrow produced Interleukin-6 with high efficiency. In addition, SK 22 stimulated macrophages to secrete tumor necrosis factor, as measured by a bioassay. Furthermore, macrophages were activated by SK 22 to produce a 3 fold amount of oxygen radicals compared to the spontaneous oxygen radical production. In contrast to this finding, the phagocytic capacity of these macrophages was only slightly increased. The specific lysis of P 815 tumor cells by peritoneal macrophages after coincubation with SK 22 was measured using tumor cells prelabelled with radioactive 51Cr. The results of the in vitro experiments presented clearly show that the E. coli preparation SK 22 is an efficient immunomodulator of the unspecific immune system.     

Inhibition of arachidonate release from rat peritoneal macrophage by biflavonoids

Biflavonoid is one of unique classes of naturally-occurring bioflavonoid. Previously, certain biflavonoids were found to possess the inhibitory effects on phospholipase A₂ activity and lymphocytes proliferation¹ suggesting their anti-inflammatory/immunoregulatory potential. In this study, effects of several biflavonoids on arachidonic acid release from rat peritoneal macrophages were investigated, because arachidonic acid released from the activated macrophages is one of the indices of inflammatory conditions. When resident peritoneal macrophages labeled with [³H]arachidonic acid were activated by phorbol 12-myristate 13-acetate (PMA) or calcium ionophore, A23187, radioactivity released in the medium was increased approximately 4.1∼7.3 fold after 120 min incubation compared to the spontaneous release in the control incubation. In this condition, biflavonoids (10 uM) such as ochnaflavone, ginkgetin and isoginkgetin, showed inhibition of arachidonate release from macrophages activated by PMA (32.5∼40.0% inhibition) or A23187 (21.7∼41.7% inhibition). Amentoflavone showed protection only against PMA-induced arachidonate release, while apigenin, a monomer of these biflavonoids, did not show the significant inhibition up to 10 uM. Staurosporin (1 uM), a protein kinase C inhibitor, showed an inhibitory effect only against PMA-induced arachidonate release (96.8% inhibition). Inhibition of arachidonate release from the activated macrophages may contribute to an anti-inflammatory potential of biflavonoidsin vivo.     

青蒿琥酯增加小鼠腹腔巨噬细胞内化内毒素、大肠埃希菌的作用及可能机制

目的:观察青蒿琥酯(artesunate,AS)对小鼠腹腔巨噬细胞内化清除脂多糖/内毒素(li-popolysaccharide/endotoxin,LPS)和吞噬大肠埃希菌的影响,并初步探讨其可能的作用机制。方法:MTT法检测不同浓度网格蛋白抑制剂(monodansylcadaverine,MDC)、内体酸化抑制剂氯喹(chloroquine,CQ)对小鼠腹腔巨噬细胞活力的影响,以选择恰当的药物工作浓度;激光共聚焦法检测青蒿琥酯及MDC、CQ对小鼠腹腔巨噬细胞内化FITC-LPS的影响;分别采用激光共聚焦和菌落集落形成计数实验观察青蒿琥酯对小鼠腹腔巨噬细胞内化大肠埃希菌的影响;逆转录PCR检测青蒿琥酯对小鼠腹腔巨噬细胞清道夫受体A(class A scavenger receptor,SR-A)mR-NA表达的影响。结果:MDC和CQ浓度分别小于25μg/mL和20μg/mL时对小鼠腹腔巨噬细胞活力无影响;MDC、CQ可抑制小鼠腹腔巨噬细胞内化LPS,青蒿琥酯可增加小鼠腹腔巨噬细胞对LPS的内化,而且青蒿琥酯可增加MDC和CQ处理的巨噬细胞内化LPS的功能。激光共聚焦和菌落集落形成计数实验均显示青蒿琥酯可增加小鼠腹腔巨噬细胞对大肠埃希菌的内化能力。逆转录PCR结果显示青蒿琥酯可增强LPS处理或未处理的小鼠腹腔巨噬细胞SR-A mRNA的表达。结论:青蒿琥酯可增强小鼠腹腔巨噬细胞内化LPS、大肠埃希菌的能力,该作用可能与SR-A mRNA表达升高有关。     

Multiplexed cytokine detection in microliter microdialysis samples obtained from activated cultured macrophages

Microdialysis sampling probes were used to collect cytokine samples from lipopolysaccharide (LPS)-stimulated macrophages. The probes were immersed into cell culture wells containing either RAW 264.7 or isolated peritoneal macrophages. Dialysates (15 microL) from these wells were subjected to a multiplexed cytokine sandwich immunoassay platform analyzed by flow cytometry that measures up to six separate cytokines, interleukin-6 (IL-6), interleukin-10 (IL-10), interleukin-12p70 (IL-12p70), interferon-gamma (IFN-gamma), macrophage chemoattractant protein-1 (MCP-1), and tumor necrosis factor-alpha (TNF-alpha) in a single 15-muL sample. In vitro microdialysis sampling relative recovery experiments showed that only IFN-gamma, IL-6, MCP-1, and TNF-alpha could be recovered across a commercially-available 100-kDa MWCO microdialysis membrane. Eleven hours after LPS addition (1 microg/mL), RAW 264.7 macrophages secreted greater than 8000 pg/mL of TNF-alpha and greater than 1000 pg/mL MCP-1. With the peritoneal macrophages, greater than 6000 pg/mL of IL-6, MCP-1, and TNF-alpha were obtained. The maximum dialysate concentrations obtained from the RAW macrophages were 1300 pg/mL for TNF-alpha and 55 pg/mL for MCP-1. Maximum cytokine concentrations from peritoneal macrophage dialysates reached approximately 2000 pg/mL, 1100 pg/mL and 500 pg/mL for TNF-alpha, MCP-1 and IL-6, respectively. Microdialysis sampling allowed for 20-min samples to be collected during the cytokine release from the activated macrophages. These results demonstrate that microdialysis sampling can be used for collection of selected cytokines with improved temporal resolution.     

吲哚美辛和美洛昔康对脂多糖诱导的小鼠腹腔巨噬细胞NF-κB的抑制作用

目的　研究非甾体抗炎药吲哚美辛(indomethacin)和美洛昔康(meloxicam)对细菌脂多糖(LPS)诱导表达的C57小鼠腹腔巨噬细胞核转录因子NF-κappaB(NF-κB)的抑制作用。方法　NF-κB的测定采用电泳迁移率改变法。结果　小鼠腹腔巨噬细胞经LPS诱导后细胞NF-κB含量明显增高。Indomethacin和meloxicam在10^-5-10^-7mol.L^-1浓度下可明显降低LPS激活的小鼠腹腔巨噬细胞NF-κB活化。结论　Indomethacin和meloxicam对NF-κB活化的抑制作用可能是两者的抗炎作用机理之一。