重组人胸腺素α原体外对IFN-γ,IFN-α及TNF-α的影响

目的　体外观察重组人胸腺素α原 (prothymosinα ,ProTα)对几种重要细胞因子分泌的影响。方法　用脾淋巴细胞、脾巨噬细胞及腹腔巨噬细胞 ,以ELISA法检测ProTα对IFN γ ,IFN α和TNF α分泌的影响。结果　 1× 10 - 7mol·L- 1 ProTα明显促进脾细胞分泌IFN γ(P <0 0 5 ) ,该浓度的ProTα也明显刺激小鼠脾巨噬细胞分泌IFN α(P <0 0 1) ;在小鼠腹腔巨噬细胞中 ,ProTα能明显刺激IFN α和TNF α的分泌 (P <0 0 1)。结论　ProTα对细胞因子IFN γ ,IFN α和TNF α的分泌均有促进作用。     

重组人胸腺素α原体外对IFN-γ、IFN-α及TNF-α的影响

目的　在体外实验中考察克隆并表达的人胸腺素α原 (prothymosinα ,ProTα)对几种重要细胞因子IFN γ、IFN α和TNF α分泌的影响。方法　在体外分离脾淋巴细胞、脾巨噬细胞及腹腔巨噬细胞 ,用ELISA法检测ProTα对IFN γ、IFN α和TNF α分泌的影响。结果　 10 - 7mol·L- 1 ProTα明显促进脾细胞分泌IFN γ(P <0 .0 5 ) ,该浓度的ProTα也明显刺激小鼠脾巨噬细胞分泌IFN α(P <0 .0 1) ;在小鼠腹腔巨噬细胞中 ,ProTα能明显刺激IFN α和TNF α的分泌 (P <0 .0 1)。结论　在体外实验中 ,ProTα对细胞因子IFN γ、IFN α和TNF α的分泌均有促进作用 ,这为ProTα进一步体内研究奠定了基础     

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。     

雷帕霉素对小鼠脾淋巴细胞走后门殖及产生细胞因子的影响

目的：研究雷帕霉素（RAPA）对ConA、PHA、LPS诱导的小鼠脾细胞增殖反应,混合淋巴细胞反应,脾T淋巴细胞亚型及细胞因子IL-2、IFN-γ、TNF-α含量的影响。方法：MTT法测定淋巴细胞增殖实验、混合淋巴细胞反应,流式细胞法测定脾T淋巴细胞亚型,结晶紫染色法测定TNF-α,E-LASI法测定IL-2,IFN-γ。结果：RAPA可显著抑制ConA、PHA和LPS诱导的小鼠脾细胞增殖和混合淋巴细胞反应（MLR）,对小鼠脾T淋巴细胞亚型无明显影响。RAPA还可明显抑制小鼠脾淋巴细胞分泌IL-2,IFN-γ,但对小鼠腹腔巨噬细胞分泌TNF-α无效。结论：RAPA抑制小鼠免疫功能的作用机制与CsA不同。     

蝎毒多肽提取物对5-Fu干预H22荷瘤小鼠免疫功能的影响

目的 探讨蝎毒多肽提取物(PESV)对5-Fu干预H₂₂荷瘤小鼠免疫功能的影响。方法 Balb/c小鼠随机分为对照组、模型组、化疗组及PESV高、低剂量组,模型组接种H₂₂肝癌,化疗组ip 20 mg/kg 5-Fu,PESV组化疗同时ig 40、10 mg/kgPESV,连续干预28 d后处死,计算瘤质量、抑瘤率、脾脏指数和胸腺指数。采用鸡红细胞吞噬实验检测H₂₂荷瘤小鼠腹腔巨噬细胞吞噬功能;流式细胞术检测小鼠脾细胞中T细胞亚群、NK细胞和NKT细胞的变化;采用ELISA法检测小鼠外周血中IFN-γ和IL-4的分泌水平。结果 与化疗组比较,PESV高剂量组瘤质量明显减轻(P < 0.05);低、高剂量组的脾脏指数、高剂量组胸腺指数、腹腔巨噬细胞吞噬率和吞噬指数、高剂量组CD4⁺T/CD8⁺T值及高、低剂量组NK/NKT细胞均显著升高(P < 0.05、0.01、0.001)。与化疗组比较,PESV高、低剂量组小鼠血清中IFN-γ水平明显增高,IL-4水平显著降低(P < 0.05、0.01)。结论 PESV可逆转5-Fu干预H₂₂荷瘤小鼠导致的免疫抑制作用。     

鲨鱼软骨粉对S_(180)荷瘤小鼠免疫细胞调节功能的影响

目的研究鲨鱼软骨粉(SCP)对S180荷瘤小鼠免疫细胞功能的影响。方法鲨鱼软骨粉300、200、100mg·kg-1小鼠灌胃给药10 d。MTT法检测其对S180荷瘤小鼠脾淋巴细胞增殖功能的影响;流式细胞仪检测T淋巴细胞亚群的变化;ELISA法测定荷瘤小鼠血清中细胞因子TNF-α、IFN-γ含量。结果 SCP能提高小鼠脾淋巴细胞的增殖活性;不同程度地提高小鼠脾细胞CD4+细胞数,增加CD4+/CD8+细胞比值;明显提高小鼠巨噬细胞的吞噬活性;促进外周血清中TNF-α和IFN-γ的分泌水平。结论 SCP能提高T细胞介导的细胞免疫应答,提高巨噬细胞调节免疫应答的功能,促进TNF-α、IFN-γ等细胞因子的分泌,从而发挥其抗肿瘤作用。     

土茯苓对环孢素A诱导的免疫抑制小鼠免疫功能的影响

目的 探讨土茯苓水提液对环孢素A诱导的免疫抑制小鼠免疫功能的影响。方法 小鼠腹腔注射环孢素A 5 d致免疫抑制模型,造模成功后随机分为模型组、左旋咪唑组（3.79 g·kg^-1,阳性药）和土茯苓低、中、高剂量组（4.55,9.1,13.65 g·kg^-1）（n=16）,连续给药治疗5 d后,处死小鼠,取脾、胸腺,称重计算脾指数和胸腺指数,流式细胞术检测CD3⁺CD4⁺,CD3⁺CD8⁺;酶联免疫吸附法检测IL-2,IL-6,IFN-γ。结果 造模后小鼠脾指数显著高于正常对照组（P<0.05）,胸腺指数明显降低,脾T细胞CD3⁺CD4⁺显著降低（P<0.05）;与模型组比较,各治疗组小鼠脾指数均有不同程度降低,土茯苓水提液中剂量组差异显著（P<0.05）,但胸腺指数无显著变化;左旋咪唑组、土茯苓中、高剂量组脾T细胞CD3⁺CD4⁺数目显著增加（P<0.01）,各治疗组脾脏IFN-γ均显著下降（P<0.05）。结论 土茯苓水提液能通过降低脾指数、增加脾T细胞CD3⁺CD4⁺的数目,增强机体免疫功能,并下调脾脏内较高的IFN-γ,其调节机制可能与左旋咪唑类似。     

毛蚶多肽提取物的体外免疫活性

目的考察毛蚶多肽提取物(PEAS)的体外免疫活性。方法 MTT法测定PEAS对小鼠脾淋巴细胞增殖的影响及对小鼠NK细胞杀伤活性的影响;中性红吞噬法测定PEAS对小鼠腹腔巨噬细胞吞噬功能的影响;流式细胞术测定PEAS所致小鼠脾淋巴细胞周期的变化;双抗体夹心ELISA法测定PEAS对主要细胞因子IFN-γ和IL-4分泌水平的影响。结果 PEAS能够显著促进小鼠脾淋巴细胞增殖(P<0.05或0.01),能够增强小鼠腹腔巨噬细胞吞噬中性红活性和小鼠NK细胞杀伤活性(P<0.05或0.01);PEAS能够促进脾淋巴细胞G0/G1期向DNA合成期(S期)转化;PEAS协同Con A作用,能够增加IFN-γ的分泌(P<0.05或0.01),并且能够抑制IL-4的分泌(P<0.05或0.01)。结论 PEAS体外可促进小鼠脾淋巴细胞、腹腔巨噬细与NK细胞的免疫功能,其机制可能与促进脾淋巴细胞DNA合成,增加IFN-γ的分泌量有关。     

壮药透骨香抗类风湿关节炎药物活性部位筛选及机制研究

目的 探讨壮药透骨香抗类风湿关节炎作用的药物活性部位及作用机制。方法 体外培养法制备BALB/c小鼠脾淋巴细胞,经刀豆蛋白A（CoA）刺激作用后,以透骨香各药物部位进行干预, 细胞计数试剂盒-8（CCK-8）法检测细胞增殖情况,计算刺激指数;体外培养小鼠脾细胞,刀豆蛋白A刺激T淋巴细胞增殖,采用酶联免疫吸附实验（ELISA）法检测观察透骨香活性部位对细胞因子白细胞介素-2（IL-2）、白细胞介素-10（IL-10）含量的影响。结果 小鼠脾细胞经刀豆蛋白A刺激后,细胞计数试剂盒-8检测光密度（OD）值与刺激指数显著升高（P＜0.05）,透骨香正丁醇部位及水溶部位能明显降低光密度值及刺激指数,其中水溶部位（5 μg·mL^-1）作用最强（P＜0.05）;刀豆蛋白A刺激脾淋巴细胞增殖后,细胞因子白细胞介素-2、白细胞介素-10分泌显著增强,透骨香水溶部位干预后能明显抑制白细胞介素-2分泌,提高白细胞介素-10含量（P＜0.05）。结论 透骨香水溶部位对T淋巴细胞增殖具有较强抑制作用,是透骨香抗RA活性部位,其机制可能与调节细胞因子白细胞介素-2、白细胞介素-10分泌有关。     

OK-432肿瘤疫苗对DBA/2小鼠脾脏细胞中Th1细胞因子以及TNF-α分泌的影响

目的利用DBA/2小鼠移植舌癌荷瘤模型,探讨OK-432肿瘤疫苗对DBA/2小鼠脾脏细胞中Th1细胞因子以及TNF-α分泌的影响。方法 ELISA定量检测脾脏淋巴细胞所产生的IFN-γ,IL-2,TNF-α等细胞因子的分泌水平。结果 OK-432肿瘤疫苗组中IFN-γ、IL-2以及TNF-α的含量明显高于实验对照组(P<0.05)。结论 OK-432肿瘤疫苗可刺激荷瘤小鼠脾脏细胞Th1细胞及TNF-α的分泌,增强DBA/2小鼠的抗肿瘤免疫功能。     

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。     

美洛昔康抑制正常人中性粒细胞与滑膜细胞粘附的作用机理研究

目的研究美洛昔康对正常人中性粒细胞(polymorphonuclear leukocyte,PMN)与滑膜细胞(human synovial cell,HSC)粘附的抑制作用及其作用机理。方法用MTT比色法检测PMN与HSC的粘附,分别用Cell-ELISA和RT-PCR法检测粘附分子ICAM-1和VCAM-1的蛋白及基因表达,用EMSA法检测NF-κB的活性。结果美洛昔康可显著的并以剂量依赖的方式抑制TNF-α(50 u·mL^-1)和IL-1β(50 u·mL^-1)作用12 h诱导的PMN与HSC粘附,其IC₅₀分别为3.38×10^-7和3.56×10^-6 mol·L^-1。进一步研究发现美洛昔康在1×10^-6～1×10^-5 mol·L^-1时还可在蛋白水平及mRNA水平抑制TNF-α(50 u·mL^-1)诱导的HSC细胞ICAM-1的表达,但对VCAM-1蛋白及mRNA表达均未见显著影响;同时美洛昔康还可显著抑制50 u·mL^-1 TNF-α诱导的NF-κB的活化。结论美洛昔康抑制NF-κB的活化,进而抑制ICAM-1的表达可能是其抑制PMN与HSC粘附的机制之一。     

氢化可的松抑制人中性粒细胞与滑膜细胞粘附机理研究

目的研究氢化可的松对正常人中性粒细胞(PMN)与滑膜细胞(HSC)粘附的作用及其机理.方法MTT比色法检测PMN与HSC的粘附,Cell-ELISA和RT-PCR法检测HSC粘附分子的表达,EMSA研究核转录因子NF-κB的活化.结果氢化可的松可显著抑制50U·mL^-1rhTNF-α与IL-1β刺激的HSC与PMN的粘附;显著抑制HSC表面VCAM-1的表达及VCAM-1mRNA表达,但对ICAM-1mRNA的表达无显著影响;同时对TNF-α诱导的NF-κB活化有显著抑制作用.结论氢化可的松显著抑制PMN与HSC的粘附,其作用机理可能是通过抑制NF-κB的活化,进而抑制滑膜细胞中VCAM-1mRNA及蛋白表达而实现的.     

苯并异硒唑酮磺酰胺衍生物对环氧酶的抑制作用

目的　研究苯并异硒唑酮磺酰胺衍生物对环氧酶的抑制作用。方法　放免法;RT-PCR法。结果　化合物A和B对COX-1和COX-2的代谢产物TXB2和PGE2的IC₅₀ 比值分别为1000和560 ,化合物A和B可抑制LPS诱导的大鼠腹腔巨噬细胞COX-2 mRNA生成,而对COX-1的mRNA生成无影响。结论　化合物A和B为COX-2选择性抑制剂,可抑制COX-2 mRNA的生成     

异丹叶大黄素对人滑膜细胞白细胞介素-8生成及mRNA表达的影响

目的研究异丹叶大黄素(isorhapotigenin,Iso)对肿瘤坏死因子α(TNF-α)诱导的人滑膜细胞(humansynovialcell,HSC)白细胞介素-8(IL-8)生成和mRNA表达的影响.方法用RIA方法测定IL-8的含量,以RT-PCR法测IL-8mRNA.结果Iso在1×10^-6-1×10^-5mol·L^-1浓度范围内对TNF-α诱导的人滑膜细胞IL-8生成有抑制作用,并抑制TNF-α诱导的HSCIL-8mRNA表达.结论Iso抑制TNF-α诱导的HSCIL-8生成,可能与影响IL-8mRNA表达有关.     

Changes in serum levels of cytokines in mice injected with an immunostimulator C3bgp isolated from Cuscuta europea

The effect of C3 binding glycoprotein (C3bgp), isolated from Cuscuta europea seeds on induction of in vivo cytokine synthesis was investigated. Different groups of mice were stimulated with 30 microg C3bgp per mouse, injected intraperitoneally. The quantitative determination of IL-1alpha, IL-2, IL-4, IL-6, IL-10, TNF-alpha and IFN-gamma was performed in mouse sera by ELISA. The quantities of these cytokines were measured at different hours: 1, 2, 3, 4, 5, 6, 7, and 24 h after injection. No significant changes in serum level of IL-2, IL-4 and TNF-alpha in experimental animal groups were found. A little increase of IL-1alpha, moderate elevation of IL-10 and IFN-gamma (5- to 6-fold more) and strong release, more than 10-fold of IL-6 in sera of C3bgp-treated mice were detected. The results obtained from C3bgp stimulated cultures of mouse peritoneal macrophages and mouse splenocytes suggest that C3bgp binds to mouse peritoneal macrophages and induces production mainly of IL-6, followed by IFN-gamma and in a very low degree of IL-1alpha and IL-10. Based on the results presented, we conclude that the increased level of IL-6 was the basic after injection of C3bgp and that the mouse macrophages were the major cell targets for the C3bgp effect.     

Immunoregulatory abnormalities of T cells and hyperreactivity of B cells in the<Emphasis Type="Italic">In Vitro</Emphasis> immune response in pristane-induced lupus mice

Systemic lupus erythematosus (SLE) is characterized by overactive B cells that differentiate into autoantibody-forming cells, aberrant T cell function that provides helping B cells produce autoantibodies, and overproduction of proinflammatory cytokines. However, immunodysregulation in lupus pathogenensis remains incomplete. We examined mitogen-stimulated production of proinflammatory cytokines, cell proliferation, T cell activation, and T cell apoptosis in vitro in pristane-induced lupus BALB/c mice compared to normal mice. LPS-stimulated production of IL-6 and IL-10 by splenocytes and macrophages from pristane-induced lupus mice were remarkably up-regulated compared to normal mice, whereas production of macrophage TNF-alpha was significantly down-regulated. Moreover, in vitro production of IL-2, IL-6, IL-10 and IFN-gamma by Con A-stimulated splenocytes, cell proliferation in LPS- or Con A-stimulated- thymocytes and splenocytes, and expression of CD69+CD4+ T cells in Con A-stimulated splenocytes were greatly increased in cells derived from pristane-induced lupus mice compared to normal mice. In addition, splenic T cells and CD4+ T cells in thymocytes from pristane-induced lupus mice were more resistant than nonautoimmune normal cells to Con A-induced apoptosis. Our findings indicate that immunoregulatory abnormalities of T cells and hyperreactivity of B cells in the in vitro immune responses in pristane-induced lupus mice may explain some of lupus pathogenesis.     

牛膝多糖的免疫调节作用

为深入探讨牛膝多糖(ABPS)对免疫系统的作用机制,用体内、外的方法,对老年鼠一些免疫功能进行了研究。研究表明,ABPS在体外可以提高老年小鼠T淋巴细胞的增殖能力和IL-2的分泌。体内能显著提高老年大鼠T淋巴细胞和血清中TNF-β或TNF-α及NO的产生和NOS的活性,降低其sIL-2R的产生;ABPS50～800mg·L^-1体外给药或100mg·kg^-1ip可提高老年大鼠PMΦTNF-α及NO的产生和NOS的活性,ABPS100mg·kg^-1ip并能提高LPS诱导的PMΦTNF-α及NO的产生和NOS的活性。ABPS对老年大鼠大脑皮层NO的产生及NOS的活性无影响。提示ABPS可以启动和活化MΦ,纠正老年鼠的免疫低下状态,是免疫调节剂。     

Effect of interleukin-4 and interleukin-10 on leucocyte migration and nitric oxide production in the mouse.

1. The effect of systemic treatment of mice with murine recombinant interleukin-4 (IL-4) or interleukin-10 (IL-10) on neutrophil infiltration into a specific tissue site and nitric oxide (NO) production from peritoneal macrophages was investigated. 2. Intravenously (i.v.) administered IL-4 (0.01-10 micrograms per mouse, approximately 0.3-300 micrograms kg-1, i.v.) and IL-10 (0.01-1 micrograms per mouse, approximately 0.3-30 micrograms kg-1, i.v.) dose-dependently inhibited neutrophil accumulation into a 6-day-old murine air-pouch induced by local application of interleukin-1 beta (IL-1 beta, 5 ng), with approximate ED50s of 0.35 and 0.90 micrograms, respectively. Neither IL-4 (1 micrograms, 30 micrograms kg-1, i.v.) nor IL-10 (1 micrograms, 30 micrograms kg-1, i.v.) prevented leucocyte accumulation in the mouse air-pouches when interleukin-8 (IL-8, 1 micrograms) was used as chemoattractant. Similarly, neither cytokine had any effect on the in vitro up-regulation of CD11b antigen on the surface of murine circulating neutrophils. 3. Treatment of mice with lipopolysaccharide (LPS, 0.3 mg kg-1, i.p.) caused an increase in the formation of NO (measured as nitrite accumulation) in the supernatant of peritoneal macrophages ex vivo. Pretreatment of mice with IL-4 (0.01-1 micrograms i.v., 20 min before LPS), but not with IL-10 (1 micrograms i.v., 20 min before LPS), caused a dose-dependent reduction in this LPS-stimulated formation of nitrite by peritoneal macrophages ex vivo. 4. Activation of murine macrophages with LPS (1 microgram ml-1 for 24 h) in vitro caused a significant increase in nitrite release in the supernatant of these cells.(ABSTRACT TRUNCATED AT 250 WORDS)     

Modulation of macrophage protease activity by acute administration of O,O,S trimethyl phosphorothioate

Previous studies showed that acute administration of O,O,S trimethyl phosphorothioate (OOS-TMP), a contaminant in malathion, acephate and fenitrothion, led to increases in metabolic activities, such as, secretion of interleukin 1 and nonspecific esterase, of splenic and peritoneal macrophages. In this report, the effect of OOS-TMP administration on the levels of the neutral proteases, elastase, collagenase and plasminogen activator, in cultures supernatants of peritoneal and splenic macrophages is presented. Acute administration of OOS-TMP elevated collagenase levels only at day 3 following treatment with 10 or 20 mg/kg OOS-TMP. Levels of elastase in culture supernatant of peritoneal and splenic macrophages, on the other hand, was elevated at days 1, 3, 5 and 7 following administration of OOS-TMP. The effect on elastase secretion was dose-dependent at days 5 and 7 after treatment. Levels of plasminogen activator activity in the culture supernatants of splenic macrophages was elevated at day 5 following treatment with both doses of OOS-TMP. At days 1 and 3, the level of plasminogen activator inhibitor was suppressed. However, at days 5 and 7 plasminogen activator inhibitory activity was close to control values. These data show that OOS-TMP administration led to an elevation in the levels of neutral proteases in culture supernatants of peritoneal and splenic macrophages. This elevation indicates that acute OOS-TMP administration alters another parameter of macrophage function, which is elevated following exposure to acute inflammatory stimuli.