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目的 探讨脂多糖(LPS)对BV2细胞肿瘤坏死因子α(TNF-α)、簇分化抗原11b(CD11b)和白细胞介素1β(IL-1β)表达的影响.方法 分别用LPS 0、1、10、100μg/ml和LPS 10μg/ml 以0、2、4、8、12、24h刺激BV2细胞,ELISA法检测细胞培养上清液中分泌的TNF-α表达量,RT-PCR 检测CD11b及IL-1β mRNA表达变化.结果 用LPS 10μg/ml刺激BV2细胞4h后,上清液中TNF-α的释放量达到峰值,同时CD11b及IL-1β mRNA表达明显增加.结论 LPS能促进BV2细胞高效表达CD11b、IL-1β和分泌TNF-α,从而为BV2细胞作为免疫活性细胞在中枢神经系统中发挥免疫调节作用提供依据. 相似文献
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目的测定基因重组人白细胞介素-Ⅱ中氨苄青霉素残留量.方法采用抗生素微生物学测定法测定基因重组人白细胞介素-Ⅱ中氨苄青霉素残留量,在测定结果为阳性时用HPLC法作氨苄青霉素鉴定.结果微生物学测定法最低检出量为0.005μ/ml,HPLC法最低检出量为0.002μg.结论本法灵敏度高,简便可行. 相似文献
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目的比较不同麻醉药物对白细胞介素(IL)-1β和IL-6的影响。方法选取健康自愿者6名,取其外周血以分层液密度梯度离心法分离出外周血单核细胞(PBMCs)。以内毒素(LPS)1μg/ml(终浓度)刺激,分为8组:①空白组(PBMCs+磷酸盐缓冲液);②损伤组(PBMCs+LPS 1μg/ml);③异丙酚组(PBMCs+LPS 1μg/ml+异丙酚4μg/ml);④氯胺酮组(PBMCs+LPS 1μg/ml+氯胺酮250μg/ml);⑤芬太尼组(PBMCs+LPS 1μg/ml+芬太尼5ng/ml);⑥咪唑安定组(PBMCs+LPS 1μg/ml+咪唑安定500ng/ml);⑦利多卡因组(PBMCs+LPS 1μg/ml+利多卡因2μg/ml);⑧布比卡因组(PBMCs+LPS 1μg/ml+布比卡因500 ng/ml)。经孵育离心后,采用酶联免疫吸附试验(ELISA)测定IL-1β和IL-6的生成。结果临床治疗浓度的异丙酚和氯胺酮对LPS刺激的PBMCs表达IL-6均有明显抑制作用,氯胺酮还能明显抑制IL-1β的表达。低浓度芬太尼、咪唑安定、利多卡因、布比卡因对IL-1β和IL-6的表达无明显影响。结论静脉麻醉药异丙酚和氯胺酮可抑制LPS刺激的PBMCs细胞因子的表达。 相似文献
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娄竞 《国际生物制品学杂志》1988,(5)
作者应用12-0-+四酰佛波醇13-乙酸(TPA)和钙离子载体A23187激活灰黄层淋巴细胞以大规模制备并纯化人白细胞介素2(IL-2)。方法如下:(1)TPA和A23187溶解于无水乙醇,浓度分别为20μg/ml及100μg/ml。(2)每一灰黄层得自献血者24小时内450ml全血,每次试验共需50个灰黄层;体积为1600~1800ml,约含1×10~(11)白细胞。(3)用血细胞洗脱仪去除灰黄层内的红细胞,11次实验去除90%的红细胞,并可使淋巴细胞与粒细胞的比率有所升高。从而解决了应用灰黄层作为大规模生产IL-2的人淋巴细胞的来源。随后用羟乙基淀粉(HES)梯度离心进一 相似文献
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咪喹莫特对致敏大鼠支气管旁淋巴结细胞培养Th1/Th2类细胞因子的影响 总被引:1,自引:0,他引:1
目的探讨免疫调节剂咪喹莫特的作用机制。方法建立致敏大鼠支气管旁淋巴结(PBLN)细胞培养体系,培养0、3、6、12、24和28h,采用酶联免疫吸附试验(ELISA)测定三种咪喹莫特浓度(0.1、1.0和10μg/ml),对培养细胞上清液中辅助性T淋巴细胞亚群Th1和Th2类细胞因子产生的影响。结果1.0μg/ml和10μg/ml的咪喹莫特对Th1类细胞因子白细胞介素(IL)-12和干扰素(IFN)-γ的产生有明显的增强作用,而对Th2类细胞因子IL-4、IL-5的产生有抑制作用,与仅有卵白蛋白(OVA)刺激组相比,差异显著(P<0.05)。此作用在培养6h开始,12h达峰值,持续至24h。结论咪喹莫特从细胞学水平即增加Th1类细胞因子产生,抑制Th2类细胞因子产生,有望为变态反应性疾病的免疫治疗提供一种新方法。 相似文献
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本文建立了一种在同一色谱条件下同时测定头孢泊肟酯干混悬剂含量及有关物质的 HPL C方法 ,并对其进行了方法学验证。以 Kromasil 10 0 -5C18(2 50 mm× 4.6mm,5μm)为色谱柱 ;0 .0 2 mol/ L乙酸铵 -乙腈 (60∶ 40 ,V/ V)为流动相 ;流速为1.0 ml/ min;检测波长 2 3 5nm;柱温为室温。结果表明 ,方法专属性良好 ;头孢泊肟酯在 12 5~ 875μg/ ml(r=0 .9995,n=7)及相关物质 2 .5~ 17.5μg/ ml (r=0 .9992 ,n=7)的浓度范围内呈线性 ;最低检出限 (L OD)为 0 .3 8μg/ ml;最低定量限 (L OQ)为 1.2 6μg/ ml;方法在高、中、低三个浓度下的平均回收率 (n=3 )及日内 (n=3 )、日间 (n=5)精密度良好 ;样品溶液于 4℃保存时 5d内稳定。本文建立的方法也适合于头孢泊肟酯原料药的分析 相似文献
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雷公藤红素对IL—1和IL—2活性及PGE2释放的抑制作用 总被引:19,自引:0,他引:19
雷公藤红素0.1~1.0μg/ml在试管内能降低LPS诱导的小鼠腹腔巨噬细胞外和细胞内白细胞介素-1(IL-1)的活性,也能抑制ConA诱导的小鼠脾细胞产生白细胞介素-2(IL-2).动态观察表明,雷公藤红素经预处理8h和3h后已能分别抑制IL-1和IL-2的产生。此外,雷公藤红素能降低A23187刺激家兔滑膜细胞释放前列腺素E_2(PGE_2)。 相似文献
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乔丽华 《国际医药卫生导报》2017,23(11)
目的 分析雾霾对慢性阻塞性肺疾病(COPD)患者呼出气冷凝液中炎性指标水平的影响.方法 选择经本院诊断为稳定期的COPD患者62例为研究对象,根据患者所住区域分为城区组(n=30例)、郊区组(n=32例),并收集患者呼出气冷凝液(EBC),检测EBC中白细胞介素(IL)-4、IL-6、肿瘤坏死因子(TNF)-α、8-异前列腺素(8-iso-PG)及白三烯B4(LTB4)水平.结果 城区组PM2.5、PM10、SO2、NO2分别为(128.56±65.32)μg/m3、(101.38±63.17) μg/m3、(76.85±44.32)μg/m3、(293.12±171.38) μg/m3,显著高于郊区组[(92.72±40.41)μg/m3、(72.53±45.32)μg/m3、(56.20±33.57)μg/m3、(202.46± 115.63) μg/m3],差异有统计学意义(均P<0.05);城区组IL-4、IL-6、TNF-α、8-iso-PG及LTB4水平分别为(6.27±1.78)pg/ml、(5.81±1.60) pg/ml、(15.43±2.82) pg/ml、(14.15±2.83)pg/ml、(48.35±9.42) pg/ml,郊区组分别为(5.37±1.72) pg/ml、(5.03±1.35) pg/ml、(14.06±2.43)pg/ml、(12.32±1.57)pg/ml、(40.32±6.47) pg/ml,城区组各指标均显著高于郊区组(均P<0.05).结论 COPD患者体内存在一定的炎性反应,当机体受到雾霾刺激后可加重炎性反应,危害机体健康,临床应对雾霾提高警惕,可通过收集患者呼出气冷凝液对病情进行监测. 相似文献
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Immunomodulatory activity of dioscorin, the storage protein of yam (Dioscorea alata cv. Tainong No. 1) tuber. 总被引:1,自引:0,他引:1
Yen-Wenn Liu Huey-Fang Shang Chung-Kwe Wang Feng-Lin Hsu Wen-Chi Hou 《Food and chemical toxicology》2007,45(11):2312-2318
The purified dioscorin from yam (Dioscorea alata L. cv. Tainong 1) tuber was previously reported (Hsu et al., 2002. J. Agric. Food Chem., 50, 6109-6113). In this report, we evaluated its immunomodulatory ability in vitro in the presence of polymyxin B (50 microg/ml) to eliminate lipopolysaccharide (LPS) contamination. Dioscorin (5-100 microg/ml) was able to stimulate nitric oxide production (expressed as nitrite concentrations) in RAW264.7 cells. The stimulation index on the phagocytosis of RAW264.7 cells against E. coli and the oxidative burst (determined by the intensity of rhodamine fluorescence) of RAW264.7 cells were both enhanced by different concentrations of dioscorin (5-100 microg/ml). The cytokine production, including IL-6, TNF-alpha, and IL-1beta in dioscorin-treated RAW264.7 cells or human monocytes, was measured in the cultured medium. Dioscorin (5-100 microg/ml) was found able to induce IL-6, TNF-alpha, and IL-1beta production in RAW264.7 cells and human monocytes. To evaluate the effects of dioscorin on the proliferation of spleen cells from BALB/c mice, phytohemagglutinin (PHA, 2 microg/ml) alone or PHA mixed with different concentrations of dioscorin (10, 25, and 50 microg/ml) was used to treat spleen cells for 24h. The stimulated proliferation index of splenic cells ranged from 1.38- to 1.48-fold of PHA alone for PHA mixed with different concentrations of dioscorin (10, 25, and 50 microg/ml). We suggest that the tuber storage protein of yam dioscorin functions as an immunomodulatory substance. 相似文献
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《Immunopharmacology》1994,27(3):207-214
Recombinant human interleukin 8 (IL-8) enhanced the release of inflammatory cytokines including interleukin 1β (IL-1β), interleukin 6 (IL-6) and tumor necrosis factor-α (TNF-α) from normal human mononuclear cells in a dose-related manner (from 1 ng/ml to 10 ng/ml with a maximal effect at 5 ng/ml( when the cells incubated with IL-8 for 24 h. This cytokine-releasing activity of IL-8 is temperature-dependent and required protein synthesis since low temperature (4°C) and cycloheximide (100 μg/ml) minimized the cytokine release from MNC. However, when IL-8 concentration was greater than 20 ng/ml, the cytokine release was supressed. For further investigating the subcellular mechanism of the adverse effect of high dose IL-8 (20 ng/ml) in cytokine synthesis, human mononuclear cells (1 × 106/ml) were stimulated with PHA (1 μg/ml) in the presence of 20 ng/ml IL-8 for 3 days. We found not only [3H]thymidine incorporation of MNC was tremendously inhibited but DNA fragmentation appeared. Subsequently, the cell cycle of PHA-stimulated MNC retarded in the phase of G0/G1. These results suggest that in low concentration (5–10 ng/ml) IL-8 not only activated neutrophil phagocytosis but facilitated the release of inflammatory cytokines from mononuclear cells. Higher dose of IL-8 (more than 20 ng/ml) conversely suppressed these cytokine release from damaged cells by its cytotoxic effect. This newly found cytokine-releasing activity of IL-8 may play a role in the modulation of inflammation. 相似文献
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人参三醇皂甙促进人白细胞介素6转译的生物学效应 总被引:2,自引:0,他引:2
将PHA激活淋巴结细胞作为研究对象,观察了人参三醇皂甙(PTGS)对人白细胞介素6(IL-6)分泌的促进效应。应用同位素示踪技术观察细胞RNA合成和蛋白质合成与IL-6分泌的关系,结合IL-6mRNA体外转译体系,探讨了PTGS促IL-6诱生的机理。结果表明PTGS对IL-6基因的转录环节无明显作用,但对IL-6基因的转译环节具明显促进效应。 相似文献
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目的 考察小豆蔻明对人淋巴细胞分泌白细胞介素-2水平的影响,探讨其作为无免疫抑制作用的细胞增殖抑制剂的可能性.方法 分离健康人静脉血淋巴细胞,制得淋巴细胞混悬液,实验细胞按正常培养及植物血凝素刺激培养后,分别加入不同药物小豆蔻明、雷帕霉素、环孢素、槲皮素、葛根素、黄豆苷元及黄芩苷进行干预,采用ELISA法测定人淋巴细胞因子白细胞介素-2的分泌水平.结果 对于植物血凝素诱导的人淋巴细胞增殖反应,低剂量小豆蔻明促进白细胞介素-2的分泌,高剂量小豆蔻明具有明显的抑制作用.结论 小豆蔻明是一种潜在的细胞增殖抑制剂,具有较大的研究和应用前景. 相似文献
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Alessandro Casini Pierangelo Geppetti Carlo Alberto Maggi Calogero Surrenti 《Naunyn-Schmiedeberg's archives of pharmacology》1989,339(3):354-358
Summary (1)We have studied the ability of some regulatory peptides to induce a mitogenic (incorporation of tritiated thymidine) response in human peripheral blood mononuclear cells (PBMC) and to modify the response produced by phytohaemagglutinin (PHA), a well known PBMC mitogen. (2) Human calcitonin gene-related peptide (hCGRP), human or salmon calcitonin (hCT, sCT), neurokinin A (NKA) and neurokinin (4–10) (up to 1 M for each peptide) did not produce per se any significant PBMC stimulation. (3) hCGRP (0.1 nM –1 M) produced a concentration dependent enhancement of the response to a submaximal concentration of PHA (1 g/ml). On the other hand, hCGRP decreased the mitogenic response to a maximal concentration of PHA (25 g/ml). (4) Neither hCT nor sCT (0.1 nM–1 M) had a significant influence on the response to PHA (1–25 g/ml). (5) Both NKA and NKA (4–10) produced a concentration-dependent (1 fM –10 pM) enhancement of the response to 1 g/ml PHA, while these compounds had no effect on the response to 25 g/ml PHA. (6) These findings suggest a potent modulatory action of CGRP and NKA, two peptides present in sensory and other nerves, on immune function which is possibly mediated via C2 receptors for CGRP and NK-2 tachykinin receptors, respectively. 相似文献
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Shin HY Jeong HJ Na HJ Kim HJ Moon G Shin TY Yang DC Hong SH Kim HM 《Biological & pharmaceutical bulletin》2005,28(7):1177-1182
Gamisanghyulyunbueum (GSHYBE) has been used clinically to treat skin related disease in South Korea. We investigated GSHYBE-mediated changes in downstream T cell signal transduction. To determine the mechanism of inhibition, we have studied many of the major pathways in phytohemagglutinin (PHA)-activated T cell. We show that among the mitogen-activated protein kinase family activation of phosphorylation of extra cellular signal-regulated kinase 1/2 (ERK1/2, p44/42) and p38, but not c-jun NH2-terminal kinase is inhibited. In activated MOLT-4 cells, the nuclear localization of nuclear factor of activated T cells (NFATc) was blocked by GSHYBE (1 mg/ml). Also, degradation of inhibitor kappaB-alpha and transactivation by nuclear factor-kappaB (NF-kappaB)/Rel A were impaired by GSHYBE (1 mg/ml). Furthermore, interlukin (IL)-2, IL-4 and Interferen (IFN)-gamma secretion by PHA activated MOLT-4 cells and peripheral blood mononuclear cells (PBMC) were significantly diminishes following GSHYBE treatment (1 mg/ml). Also, oral administration of GSHYBE inhibited IL-2 secretion in skin allergic reaction. In conclusion, our data indicate that GSHYBE treatment of T cells inhibits ERK1/2 and p38 activation and nuclear translocation of NFATc, NF-kappaB, resulting in diminished secretion of IL-2. 相似文献
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The effect of the environmental pollutants, diesel exhaust particles (DEP) and formaldehyde (FA), on the production of pro-inflammatory cytokines (interleukin (IL)-1alpha, IL-1beta, tumor necrosis factor (TNF)-alpha and IL-8) by normal human dermal keratinocytes (hKCs) was investigated. Normal hKCs were incubated with various concentrations of DEP (0.4, 0.8, 4, or 20 microg/ml) or FA (0.25, 0.5, 1, or 5 microg/ml), and cytokine production was then determined by enzyme-linked immunosorbent assay (ELISA). DEP (20 microg/ml) induced IL-1beta production without altering cell growth. The increased production of IL-1beta induced by this concentration of DEP was further enhanced by the presence of phorbol 12-myristate 13-acetate (PMA), although PMA alone did not affect the levels of IL-1beta. IL-8 production was also increased by DEP (0.4 and 0.8 microg/ml), which is consistent with the results that these concentrations of DEP increased the number of cells significantly after 72 h incubation. Although FA alone did not stimulate the production of IL-1beta or IL-8 by keratinocytes, FA (0.5 microg/ml and 5 microg/ml) significantly increased IL-8 and IL-1beta production, respectively, in cells stimulated with PMA. IL-1alpha production was not modulated by FA or DEP even in the presence of PMA. TNF-alpha was produced by unstimulated keratinocytes at barely detectable levels after 48 h incubation. Although basal levels of TNF-alpha in the culture supernatants were increased after stimulation with PMA, neither pollutant alone nor combination with PMA affected the levels of TNF-alpha. These in vitro findings suggest that environmental pollutants may act as modulating factors of cutaneous inflammation by affecting the ability of keratinocytes to release pro-inflammatory cytokines. 相似文献
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R D Maca 《Immunopharmacology》1983,6(4):267-277
The effects of PGE2 on cultured T lymphocytes (CTC) stimulated by either Con A, PHA, or lectin-free IL-2 were studied. PGE2, in a concentration ranging from 100 to 1 ng/ml, consistently and significantly inhibited the proliferation of CTC induced by either PHA or Con A. PGF2 alpha was essentially without effect. Although the degree of inhibition of PHA-treated CTC was increased with suboptimal amounts of PHA, significant inhibition still resulted with optimal PHA concentrations. PGE2, but not PGF2 alpha, was also effective in significantly inhibiting the proliferation of IL-2-treated CTC in a dose-related fashion; however, the addition of suboptimal amounts of IL-2 did not result in greater increases in the degree of inhibition by PGE2. Depleting the CTC of either OKT-4 or OKT-8 phenotypic cells did not abrogate this PGE2 inhibitory effect, indicating that PGE2 does not suppress the proliferative response solely by the activation of suppressor cells with the OKT-8 phenotype. PGE2 also was found to inhibit the production of IL-2 by fresh lymphocytes treated by either optimal or suboptimal amounts of PHA, however, this decrease in production by PGE2 was not necessarily associated with a decrease in the proliferation of these stimulated lymphocytes. Only with low PHA concentrations, where IL-2 production was markedly reduced and barely detectable, was lymphocyte proliferation appreciably reduced by PGE2. In additional experiments, LiCl was added to PGE2 containing cultures to determine whether LiCl could modulate the inhibitor effect of PGE2 of either PHA- or IL-2-stimulated CTC. In these studies, LiCl, in concentrations of 1-10 mM was found to lessen or completely abrogate the reduced PHA proliferative response induced by PGE2. This effect was more pronounced with suboptimal concentrations of PHA than with optimal PHA amounts. In contrast, the PGE2-induced inhibition of IL-2-stimulated CTC was not modified or altered by the addition of LiCl. Thus, these results suggest that LiCl acts at the level of IL-2 production instead of IL-2 action, and that PGE2 inhibits IL-2-induced proliferation of CTC by a different or additional mechanism than for PHA-treated cells. In conclusion, these results, taken as a whole, indicate that PGE2 suppresses the proliferation of stimulated CTC by at least two different mechanisms: 1) by reducing the production of IL-2 by stimulated lymphocytes; and 2) by directly acting on the responding CTC.(ABSTRACT TRUNCATED AT 400 WORDS) 相似文献
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Zidovudine (ZDV), an anti-human immunodeficiency virus (HIV) therapy, has been associated with reduction in mortality and improvement of patients with acquired immunodeficiency syndrome (AIDS). The ZDV recipients, however, experience a multitude of side effects of which bone marrow suppression is the most noteworthy, especially among patients with low CD4 cell counts. The effect of ZDV and interleukin-2 (IL-2) on phytohemagglutinin (PHA)-induced proliferative response of peripheral blood mononuclear cells (PBMs) from patients with HIV infection was investigated. ZDV 0.5 micrograms inhibited 40% of PHA-induced thymidine uptake in PBMs from healthy donors or patients with HIV, irrespective of their CD4 cell counts. However, IL-2 (10 U/ml) had differential effect on PHA-induced thymidine uptake that appeared to be dependent on absolute CD4 cell counts. While PBMs from patients with CD4 cell counts of 400/mm3 or more did not respond to IL-2 (low responders), IL-2 enhanced the PHA-induced thymidine uptake in PBMs from patients with CD4 cell counts less than 400/mm3 at an average of 60% (high responders). Moreover, IL-2 restored the ZDV-induced inhibition by almost 100% in the high responder group while it did not affect counts in the low responder group. The production of IL-2 in vitro, in response to PHA or recall antigens, was equivalently inhibited in both groups. These data suggest that ZDV and IL-2 could have an additive effect on immune parameters in certain groups of patients infected with HIV. The differential effect of IL-2 was independent of IL-2 receptor expression. 相似文献