脑梗死患者血清白介素-6、白介素-8、肿瘤坏死因子-α及白介素-1β水平的研究

目的研究脑梗死患者急性期血清 IL -6、IL-8、TNF-α、IL -1β水平的变化 ,特别是 IL -6水平与脑梗死面积及神经功能缺损的关系。方法采用双抗体夹心 ELISA法检测 3 4例脑梗死急性期患者不同时间内血清 IL-6、IL -8、TNF-α、IL -1β水平 ,并随机抽取 10例健康人进行对照。结果脑梗死患者血清 IL -6水平在发病 6小时内明显升高 ,2 4～ 3 6小时达高峰 ,7天后基本降至正常 ,大面积梗死组IL-6水平明显高于小面积梗死组 ( 6小时内 ,P <0 .0 5 ;2 4～ 3 6小时 ,P <0 .0 5 ) ,且血清 IL -6水平与神经功能评分呈正相关( r=0 .87,P<0 .0 1) ,七叶皂甙钠治疗 3天后血清 IL-6水平下降明显 ( P<0 .0 1) ,脑梗死患者急性期血清 IL-8水平亦升高( P<0 .0 5 ) ,TNF-α、IL-1β水平则无明显变化。结论急性脑梗死患者血清 IL-6水平升高 ,而 IL-6升高的水平反映应激反应程度 ,与梗死面积及神经功能缺损程度有一定相关性。     

硫酸镁对大鼠急性颅脑损伤后白介素-1β表达的影响

目的：研究硫酸镁对急性颅脑损伤后白介素－1β（Inter leukin-1β,IL－1β）表达的影响,探讨镁离子保护创伤神经元的可能机理。方法：建立大鼠脑外伤模型,采用免疫组化方法观察镁离子对颅脑损伤后IL－1β表达的影响。结果：实验大鼠大脑局部损伤及区及周围神经细胞中IL－1β阳性产生物表达明显增加,伤后4h,8h,12h较1h升高非常显著（P＜0．01）;伤后给予硫酸镁可以抑制IL－1β的过高表达,其组织学中神经组织变性、坏死及血管炎性反应也明显减轻。结论：颅脑损伤后,受损脑组织中IL－1β表达增加,硫酸镁可以通过抑制后IL－1β表达,起到保护创伤神经元的作用。     

慢性血液透析患者血清白介素-1β、白介素-6及肿瘤坏死因子-α变化及意义

血液透析(HD)通过血液、透析膜、透析液间相互反应可激活外周单个核细胞并影响到机体细胞因子产生,而细胞因子白细胞介素-1β(IL-1β)、白细胞介素(IL-6)为炎性介质,在致热原反应中起一定作用,同时已知IL-6、肿瘤坏死因子-α(TNF-α)是导致肾小球基质增生、系膜细胞增殖、肾小球硬化及病变进展、肾功能恶化重要因素之一.此项研究采用酶免疫吸附(ELISA)双夹心抗体法测定30例健康对照组,30例慢性肾功能不全(CRF)未血液透析及60例CRF血液透析患者透析前后白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)及肿瘤坏死因子-α(TNF-α)水平.并检测HD患者第一次血液透析时,单次透析前后上述指标改变,以探讨CRF患者细胞免疫功能的改变及透析对其影响.     

白介素-1β及其受体拮抗蛋白在肠缺血-再灌注所致急性肺损伤中的作用研究

目的：探讨肠缺血－再灌注（I／R）损伤时肺损伤的机制和白介素－1β及其受体拮抗蛋白（IL－1ra)的作用。方法：用大鼠肠系膜上动脉夹闭造成I／R损伤模型,利用放射免疫分析法测定损伤前后和各处理组血液、肺组织灌洗液中IL－1β和IL-1ra水平。同时采用逆转录－聚合酶链反应（RT－PCR）法测定肺组织中IL－1β和IL－1ra mRNA的表达含量。经光镜和电镜观察肺局部白细胞浸润及组织损伤情况。结果：I／R损伤后6小时血液中IL－1β和IL－1ra有所增加,损伤组IL－1β比对照组明显增加,而IL-1ra不明显;但损伤组和对照组IL－1ra伤后6小时比伤前自身对照组均明显增加。损伤组肺灌洗液中IL－1β明显增多,而L－1ra不明显。肺组织中IL－1β的mRNA在损伤后表达明显增加,而IL－1ra表达没有明显变化。经非特异性磷脂酶A2（PLA2）阻断剂（氯喹和三氯拉嗪）、血小板活化因子受体阻断剂SR27417A、环氧化酶抑制剂消炎痛以及抗氧化剂愈创木酸等处理后,血清、肺灌洗液和肺组织中上述指标有不同程度的改变。肺内白细胞的聚集和浸润明显减少。结论：IL－1β和IL－1ra可能参与肠I／R介导的急性肺损伤,并且和局部PLA2活化有一定关系。     

格雷夫斯病患者外周血IL—1β和IL—6的测定及意义

目的 探讨格雷夫斯病（Graves病，简称GD）时外周血IL－1β和IL－6的变化及其相关关系。方法 采用双抗体夹心ELISA法测定Graves病患外周血IL－1β及IL－6的水平。结果 GD组血清IL－1β及IL－6水平与正常对照组比较均显增多（P＜0．01），IL－1β与IL－6呈显正相关关系。结论 IL－1β和IL－6在Graves病的发生发展过程中起了重要作用。     

养血清脑颗粒对偏头痛患者血中白介素—1β的影响研究

目的 探讨白介素－1ββ（IL－1β）在偏头痛发病机制中的作用及养血清脑颗粒对血中IL－1β的影响。方法 分别检测40例正常健康人及40例偏头痛患者治疗前后血中白介素－1β浓度。结果 偏头痛患者发作期血中IL－1β浓度（25．09&;#177;15.17)pg/ml，较正常健康人组高（8．50&;#177;3．50）pg/ml(P&;lt;0.001)，养血清脑颗粒治疗前后偏头痛患者血中IL－1β浓度明显降低（9．52&;#177;3.15)pg/ml(P&;lt;0.001)。结论 研究说明IL－1β在偏头痛发作时起着炎症介导作用，同时说明养血清脑颗粒可能有抑制偏头痛发作时的炎症反应。     

急性脑出血患者脑脊液中白介素-6及可溶性白介素-6受体的检测

白介素 6 (IL 6 )被认为在中枢神经系统中具有神经保护和神经营养作用 ,其重要性倍受关注。有研究报道 ,在脑出血患者外周血单核细胞及脑脊液中IL 6的表达显著增高〔1 ,2〕,而对脑出血患者脑脊液 (CSF)中可溶性 IL 6受体(s IL 6 R)的变化尚未见报道。我们对急性脑出血患者 CSF中 IL 6和 s IL 6 R进行联合测定 ,以了解其临床意义及两者的相互关系。报告如下。1　资料与方法1.1　病例 :1999年 1月— 2 0 0 1年 8月我院神经外、内科收治的经临床和 CT证实的自发性脑出血患者 2 5例 ,男 13例 ,女 12例 ;年龄 12～ 75岁 ,平均 49岁。脑…     

血清白介素1β，白介素6，白介素8，白介素10和肿瘤坏死因子α参考区间的初步建立

目的初步建立本实验室Immulite 1000分析系统检测血清白介素1β(IL-1β)、白介素6(IL-6)、白介素8(IL-8)、白介素10(IL-10)和肿瘤坏死因子α(TNF-α)的参考区间。方法收集表观健康成年人的血清,采用化学发光免疫分析检测血清中的IL-1β、IL-6、IL-8、IL-10和TNF-α水平。结果 IL-1β的95%参考区间为5 pg/mL,IL-6为2.95 pg/mL,IL-10为5 pg/mL。IL-8的95%参考区间男性85.3 pg/mL,女性67 pg/mL;TNF-α的95%参考区间男性10.96 pg/mL,女性9.77 pg/mL。结论初步建立了本实验室的血清IL-1β、IL-6、IL-8、IL-10和TNF-α在Immulite 1000分析系统的参考区间。     

CAPD患者腹腔白介素-1β和前列腺素变化及意义

为了探讨持续非卧床腹膜透析(CAPD)患者在伴发腹膜炎情况下腹腔透出液(PDE)中白介素1β(IL-1β)和前列腺素变化及意义,对17例伴发腹膜炎的CAPD患者进行了从炎症发作至消退(对照)连续10日的随访.结果显示,在炎症发作当日,PDE中IL-1β,血栓素B2(TXB2)和6-酮-前列环素(6-keto-PGF1α)均明显升高(112.8±23.40vs13.3±3.42pg/ml、370.5±52.37vs121.5±42.31ng/ml、378.9±21.66vs40.3±11.78ng/ml,P＜0.01).IL-1β于第2日迅速下降至23.9±4.56pg/ml,第2日后与对照无差异(P＜0.05).TXB2与6-keto-PGF1α于第2～4日逐渐下降,至第5日以后与对照无差异(P＞0.05),其变化过程与腹膜炎的恢复基本同步.提示IL-1β的介导CAPD患者腹膜急性炎症反应中有重要作用.而前列腺素的延迟恢复可能与腹膜的炎症防御作用有关.     

白介素6对人Th17细胞的免疫调节作用

本研究旨在探讨白介素6(IL-6)对人Th17细胞的调节作用.应用免疫磁珠分离正常人CD4+ T细胞并培养.实验分为2组,实验组(IL-6+):CD4+ T细胞(1×106/ml)经重组人IL-6(20ng/ml)刺激4天;对照组(IL-6-):不经IL-6刺激.应用酶联免疫吸附试验(ELISA)检测上清中IL-17蛋白水平,流式细胞术(FCM)检测Th17细胞数量.结果发现,与对照组相比,经IL-6刺激后的CD4+ T细胞上清中IL-17蛋白水平明显升高(337.05±189.09pg/ml vs 15.07±12.70 pg/ml)(p<0.05);进一步FCM发现,IL-6剌激组Th17细胞数量明显高于对照组[(4.05± 0.30)% vs(2.81±0.44)]%,(p<0.01).结论:IL-6促进人Th17细胞的增殖.     

The increased release of prostaglandin E2 by Kupffer cells from burned guinea pigs

Prostaglandin E2 (PGE2) is a very important immunosuppressive substance that is synthesized and released by all macrophages including Kupffer cells (KCs). In this study the changes of PGE2 released by KCs were evaluated in a burned guinea pig model. Prostaglandin E2 was released by KCs from burned guinea pigs at a consistently and significantly high level when stimulated with endotoxin. On postburn day 8, KCs that were cocultured with hepatocytes released significantly less PGE2. There was also a significant decrease in antibody-dependent cell-mediated cytotoxicity on postburn day 1. Kupffer cells can mediate immune suppression in burn injury by a prolonged increased production of PGE2 that has immunosuppressive effects on other cells and by a decreased cytotoxic effect soon after burn injury.     

Glycyrrhizin prevents liver injury by inhibition of high-mobility group box 1 production by Kupffer cells after ischemia-reperfusion in rats

High-mobility group box 1 (HMGB1) acts as an early mediator of inflammation and organ damage in hepatic ischemia-reperfusion (I/R) injury. Glycyrrhizin is a natural anti-inflammatory and antiviral triterpene in clinical use. The purpose of this study was to investigate the effect of glycyrrhizin on liver injury caused by I/R and production of HMGB1 by Kupffer cells in rats. In the first test period, rats were given saline or glycyrrhizin 20 min before segmental hepatic warm I/R. Serum alanine aminotransferase and HMGB1 levels and hepatic histopathological findings were evaluated after I/R. Furthermore, expression of HMGB1 in the liver was assessed by immunohistochemical staining after I/R. Kupffer cells were isolated by collagenase digestion and differential centrifugation, and production of HMGB1 was assessed. In another set of experiments, the effect of inhibition of Kupffer cells by injection of liposome-entrapped dichloromethylene diphosphonate (lipo-MDP) on liver injury and expression of HMGB1 were investigated after I/R. Liver injury was prevented in the glycyrrhizin group compared with the control group. Furthermore, serum HMGB1 levels were also significantly blunted in the glycyrrhizin group compared with the control group. Cells expressing HMGB1 were detected in the hepatic sinusoid by immunohistochemistry and recognized morphologically as Kupffer cells. Furthermore, the expression of HMGB1 was reduced in the glycyrrhizin group compared with the control group. Production of HMGB1 was reduced in Kupffer cells isolated from the glycyrrhizin group compared with the control group. It is noteworthy that treatment with lipo-MDP significantly blunted serum HMGB1 levels and prevented liver injury after I/R. These results suggest that glycyrrhizin has the therapeutic potential to prevent warm I/R-induced injury during hepato-biliary surgery.     

Differential effect of anti-TNF-alpha antibody on proinflammatory cytokine release by Kupffer cells following liver ischemia and reperfusion.

To study the role of tumor necrosis factor-alpha (TNF-alpha) for induction of the proinflammatory cytokine cascade after liver ischemia and reperfusion (I/R), rats were injected intraperitoneally with anti-TNF-alpha monoclonal antibodies (mAb) or placebo (IgG1) 30 min prior to global hepatic ischemia. Blood levels of TNF-alpha, interleukin (IL)-1alpha and -6 were determined. In addition, Kupffer cells (KC) were harvested after 60 min of reperfusion and spontaneous cytokine release was measured. Sham-operated animals were used as controls. Levels of proinflammatory cytokines in serum and KC supernatants were detected using specific bioassays and ELISA. Liver I/R resulted in increased (p < .01) serum levels of TNF-alpha, IL-1alpha, and IL-6, which was associated with an enhanced (p < .05) release of these cytokines by KC. In vivo pretreatment with anti-TNF-alpha mAb led to complete neutralization of TNF-alpha serum levels and decreased (p < .01) IL-6 levels (-62%). Moreover, anti-TNF-alpha mAb markedly (p < .05) decreased the release of TNF-alpha (-69%) and IL-6 (-56%) by KC, while IL-1alpha was not affected. These data indicate that TNF-alpha produced early after liver I/R triggers both its own secretion as well as IL-6 release by KC during reperfusion while the release of IL-1alpha occurs independent from TNF-alpha.     

Activated neutrophils induce nitric oxide production in Kupffer cells

Neutrophils (PMN) are proposed to contribute to hepatic dysfunction during sepsis. Transmigrating PMN have been demonstrated to adhere to and injure parenchymal cells (hepatocytes); however, the effect of sepsis-activated PMN on hepatic macrophages or Kupffer cells (KC) is poorly characterized. We hypothesize that PMN influence KC inflammatory mediator production, including nitric oxide. Rodent KC were co-cultured with PMN obtained from controls (Norm-PMN) or endotoxemic rats [lipopolysaccharide (LPS)-PMN] for 18 h. After an 18-h incubation, supernatants and cell lysates of the KC were analyzed for nitric oxide (NO) production. Co-cultures with LPS-PMN/KC demonstrated significantly increased production of nitrite and up-regulation of inducible nitric oxide synthase (iNOS) protein compared to KC alone or Norm-PMN/KC co-cultures. Immunohistochemistry revealed preferential iNOS protein staining in the cytoplasm of KC cultured with LPS-PMN compared to controls. Nitrite production in co-cultures of KC and LPS-PMN where cell contact was inhibited by a cell impermeable but diffusable membrane was significantly reduced. These data provide evidence that KC can be stimulated directly by activated PMN for production of NO. Further, they suggest another mechanism by which PMN modulate hepatic function during sepsis.     

NMDA受体过度激活对严重创伤大鼠血清炎性细胞因子水平的影响

摘要：目的 观察N-甲基-D-天冬氨酸（N-methyl-D-aspartate， NMDA）受体的过度激活对大鼠严重创伤血清炎性细胞因子水平中的影响，为探索从中枢的某一环节着手，来抑制严重创伤后的炎性反应失调提供理论依据。方法 以30% TBSA Ⅲ度烧伤为严重创伤模型，利用ELISA方法检测激活NMDA受体对血清炎性细胞因子TNF-α、IL－1β、IL6水平的影响；通过膜片钳技术检测严重创伤能否导致大鼠神经元NMDA受体的过度开放；再观察阻断NMDA受体能否抑制严重烧伤后血清炎性细胞因子TNF-α、IL－1β、IL6水平的上升。结果 ①与对照组相比，使用NMDA 0.5mg/kg激活NMDA受体，血清TNF－α，IL－1β、IL6明显升高，加大剂量（2mg/kg ）可以使血清TNF－α，IL－1β、IL6进一步升高；②在35pS电导水平的开放中，烧伤使通道开放概率增加非常显著，在100pS电导水平的开放中，开放时间常数τ1、通道开放概率增加非常显著；③腹腔注射MK-801（3mg/kg）阻断NMDA受体可以抑制烧伤后血清TNF-α、IL－1β、IL6水平的上升，加大注射剂量（5mg/kg）可以进一步抑制烧伤后血清TNF-α、IL－1β、IL6水平的上升。结论 NMDA受体是严重创伤后（烧伤）大鼠血清炎性细胞因子过度升高的重要环节。     

Spontaneous production of tumor necrosis factor alpha by Kupffer cells of MRL/lpr mice

We report that freshly isolated, unstimulated Kupffer cells (KC) from MRL/lpr female mice in short-term culture spontaneously produce high levels of TNF-alpha. TNF production was first detected in KC cultures at age 6 wk and increased with the age of the mice. Moreover, the levels of spontaneous TNF production by KC directly correlated with the age of the MRL/lpr mice. Although TNF production by KC could be induced with C. parvum in vivo or LPS in vitro in all nonautoimmune C3H/HeN, BALB/c, DBA/2, C57B16 mice, the only other strain in which spontaneous TNF production by KC was observed was MRL/++ mice greater than 10 mo old.     

Involvement of Kupffer cells in the interaction between neutrophils and sinusoidal endothelial cells in rats

During endotoxic liver injury, large numbers of neutrophils infiltrate the liver, and serum levels of tumor necrosis factor-alpha (TNF-alpha) become elevated. The object of this study was to assess the roles of TNF-alpha secreted by Kupffer cells in the interaction between neutrophils and sinusoidal endothelial cells (SECs). Rat neutrophils were perfused onto SECs that were stimulated with either TNF-alpha or supernatant from lipopolysaccharide (LPS)-stimulated Kupffer cells using an in vitro flow system. Numbers of adhered or migrated neutrophils were counted, and the effect of an antibody against intercellular adhesion molecule-1 (ICAM-1) was studied. Compared with controls (200 +/- 21 cells/mm2), neutrophil adhesion to SECs was significantly increased by both TNF-alpha (342 +/- 26 cells/mm2; P < 0.05) and LPS-stimulated Kupffer cell supernatant (331 +/- 29 cells/mm2; P < 0.05). Anti-ICAM-1 significantly inhibited neutrophil adhesion (139 +/- 10 cells/mm2; P < 0.05) and decreased the migration rate of neutrophils on SECs treated with LPS-stimulated Kupffer cell supernatant (P < 0.05). LPS-stimulated Kupffer cells secreted TNF-alpha in an LPS dose-dependent manner, and they significantly enhanced ICAM-1 expression on SECs (P < 0.05 vs. control). In addition, dexamethasone suppressed TNF-alpha production by LPS-stimulated Kupffer cells and decreased ICAM-1 expression and neutrophil adhesion on SECs. These findings suggest that Kupffer cells are involved in neutrophil adhesion and migration in hepatic sinusoids via TNF-alpha production and induction of ICAM-1 expression on SECs during liver injury associated with endotoxemia.     

早期药膳饮食对严重烧伤大鼠肠道细菌内毒素移位的影响

目的 探讨早期药膳饮食对严重烧伤早期肠道细菌内毒素移位的影响,探索保护胃肠黏膜,减少肠道细菌移位,预防脓毒症的发生方法 .方法 将健康Wistar大鼠100只,随机分成烧伤药膳喂养组(药膳组,n=30)、烧伤肉汤喂养组(肉汤组,n=30)、烧伤常规饲养组(常规组,n=30)和正常对照组(对照组,n=10).药膳组、肉汤组、常规组于伤后第1、3、7天各取10只,无菌条件下取材检测细菌移位率、血浆内毒素含量.结果 与对照组比较,药膳、肉汤及常规组细菌移位率伤后第1、3天明显升高(P<0.01或P<0.05);血浆内毒素水平烧伤后各时间点均明显升高(P<0.01).与肉汤、常规组比较,药膳组细菌移位率及内毒素水平伤后各时间点均下降(P<0.01或P<0.05).结论 早期应用药膳饮食明显降低了严重烧伤大鼠的肠道细菌和内毒素移位,表明其对肠道屏障功能有保护作用.     

抑制枯否细胞对脓毒症大鼠肝脏微循环的影响

目的 探讨抑制枯否细胞(Kupffer cells,KCs)对脓毒症大鼠肝脏微循环的影响.方法 采用盲肠结扎穿孔法(CLP)制备大鼠脓毒症急性肝损伤模型.造模前1 d和2 d经尾静脉注射三氯化钆(GdC13)去除KCs.将60只健康SD大鼠随机分为四组,即对照组、假手术组(Sham组)、模型组(CLP组)和GdC13组(CLP+GdC13组).各组动物于术后24 h处死.检测血中丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、内毒素(ET)、内皮素-1(ET-1)及一氧化氮(NO)的水平.提取肝组织RNA,采用RT-PCR方法检测ET-1、eNOS、iNOS及HO-1 mRNA的表达.结果 与对照组比较,脓毒症大鼠血中ALT、AST、ET、ET-1及NO水平显著升高(P<0.05);去除KCs后,脓毒症大鼠血中ALT、AST、ET、ET-1及NO水平显著降低(P<0.05).脓毒症大鼠肝组织ET-1、eNOS、iNOS及HO-1 mRNA 表达水平较对照组显著升高(P<0.05);去除KCs后,肝组织中iNOS及HO-1 mRNA表达水平显著降低(P<0.05).结论 KCs在脓毒症时肝脏的微循环障碍中起重要作用.