糖皮质激素对肿瘤坏死因子引起的大鼠中性粒细胞粘附的影响

采用中性粒细胞与玻璃珠粘附和ＰＭＮ与血管内皮细胞粘附两种模型，以肿瘤坏死因子，作为ＰＭＮ的刺激因子，研究糖皮质激素对ＴＮＦ引起的大量ＰＭＮ粘附的影响，同时给予糖皮质激素受体阻断剂ＲＵ３８４８６观察ＧＲ在粘附中的作用，结果发现，ＴＮＦ能明显增强大鼠ＰＭＮ的粘剂；Ｄｅｘ不能抑制经ＴＮＦ预处理的ＰＭＮ的粘附（Ｐ＞０．０５），但有一定的预防作用；经ＴＮＦ预处理再同时给予Ｄｅｘ和ＲＵ３８４８６的ＰＭＮ粘附同     

糖皮质激素受体阻断对烫伤大鼠肺、肾血管壁通透性变化的影响

我们以糖皮质激素受体（ＧＲ）的竞争性拮抗剂ＲＵ３８４８６（简称ＲＵ４８６）阻断大鼠体内的ＧＲ，并通过检测肺、肾组织匀浆中荧光标记白蛋白的含量，观察了烫伤后１２ｈ大鼠肺、肾血管壁通透性的变化以及阻断ＧＲ对这种变化的影响。结果显示：烫伤后１２ｈ，大鼠肺、肾组织匀浆中荧光标记白蛋白含量明显高于对照组（肺：Ｐ＜０．０５；肾：Ｐ＜０．００１）；阻断ＧＲ后再烫伤大鼠，其肺、肾组织匀浆中的荧光白蛋白含量又显著高于烫伤组（Ｐ＜０．０５）。提示：①烫伤后１２ｈ，大鼠肺、肾血管壁通透性明显升高。②ＧＲ减少可加重烫伤所致的血管壁通透性升高，并可逆转内源性糖皮质激素（ＧＣ）稳定血管壁通透性的作用。     

糖皮质激素受体阻断对烫伤大鼠肺‘肾血管壁通透性变化…

我们以糖皮质激素受体的竞争性拮抗剂ＲＵ３８４８６？（简称ＲＵ４８６）阻断大鼠体内的ＧＲ，并通过检测肺、肾组织匀浆中荧光标记白蛋白的含量，观察了烫后１２ｈ大鼠肺、肾血管壁通透性的变化以及阻断ＧＲ对这种变化的影响。结果显示：烫伤后１２ｈ，大鼠肺、肾组织匀浆中荧光标记白蛋白含量明显高于对照组（肺：Ｐ＜０．０５；肾：Ｐ＜０．００１）；阻断ＧＲ后再烫伤大鼠，其肺、肾组织均浆中的荧光白蛋白含量又显著高于烫伤组     

慢性肾衰患者血红蛋白浓度变化对血清细胞因子活性的影响作用

目的为了探讨贫血与慢性肾衰（ＣＲＦ）免疫功能状态的关系。方法检测了ＣＲＦ患者单纯输血及用红细胞生成素（ＥＰＯ）治疗前后细胞因子白细胞介素２（ＩＬ２）、可溶性白细胞介素２受体（ｓＩＬ２Ｒ）、肿瘤坏死因子（ＴＮＦ）及γ干扰素（γＩＦＮ）水平，并与肾功能正常肾小球肾炎患者（ＧＮ）组及对照组（Ｃ）进行比较分析。结果ＣＲＦ组血清ＩＬ２，ＴＮＦ和γＩＦＮ水平均显著低于ＧＮ组及Ｃ组（Ｐ＜００１），ｓＩＬ２Ｒ水平则较ＧＮ组及Ｃ组明显升高（Ｐ＜００１）。ＣＲＦ组血清ＩＬ２，ＴＮＦ和γＩＦＮ水平与血红蛋白浓度存在直线正相关，而ｓＩＬ２Ｒ水平与血红蛋白浓度呈负相关性，单纯输血或应用ＥＰＯ治疗能改变这些细胞因子活性水平。结论ＣＲＦ免疫功能低下与贫血有关，及时治疗和改善贫血状态可部分纠正这种免疫异常。     

肾病综合征儿童淋巴细胞糖皮质激素受体测定

用放射配基受体结合法测定淋巴细胞糖皮质激素受体（ＧＣＲ）。３３例健康儿童的均值为３５２７±２１６１，２３例肾病综合征（ＮＳ）儿童的均值为１３６５８±１３６６５．Ｐ＜０．０１。１７例ＮＳ患儿淋巴细胞ＧＣＲ≥正常值，其中１２例对激素治疗敏感。６例ＮＳ患儿淋巴细胞ＧＣＲ＜正常值，３例对激素不敏感，３例早期复发，用淋巴细胞ＧＣＲ预测ＮＳ患儿对激素治疗反应的符合率为７８％。     

小儿肾脏疾病血,尿内皮素的变化

目的研究小儿肾脏疾病血、尿内皮素（ＰＥＴ、ＵＥＴ）的水平及其相互关系。方法采用同位素放免方法检测了肾病综合征（ＮＳ），肾小球肾炎（ＧＮ），肾功能衰竭（ＲＦ）共７２例患儿血及尿中ＥＴ，血心钠素（ＡＮＰ）水平。结果ＮＳ，ＧＮ，ＲＦ三组的ＰＥＴ及ＵＥＴ值明显高于对照组，尤其ＲＦ组（Ｐ＜００５，Ｐ＜０．０１）。ＡＮＰ值在ＧＮ组和ＲＦ组明显高于对照组（Ｐ＜００１）。８例ＡＲＦ患儿恢复期血ＥＴ水平下降，６例ＣＲＦ患儿虽经治疗，但血ＥＴ水平不降或上升。结论ＥＴ在小儿肾脏疾病发病机理及病情进展中可能起重要作用，其值高低与病情严重程度及预后有关。     

地塞米松对创伤性急性肺损伤家兔肿瘤坏死因子-α的干预作用

目的 探讨地塞米松（Ｄｅｘ）对创伤性急性肺损伤（ＡＬＩ）治疗作用的可能机制。方法 采用逆转录聚合酶链反应（ＲＴ－ＰＣＲ）、酶联免疫吸附法（ＥＬＩＳＡ）检测２４只大耳白兔肺组织肿瘤坏死因子－α基因（ＴＮＦ－αｍＲＮＡ）表达及肺泡巨噬细胞（ＡＭ）培养上清液中肿瘤坏死因子－α（ＴＮＦ－α）、白细胞介素（ＩＬ）－６水平。结果 创伤性ＡＬＩ兔肺组织ＴＮＦ－αｍＲＮＡ表达及ＡＭ培养上清液中ＴＮＦ－α,ＩＬ－６含量较正常对照组比较明显升高（Ｐ〈０．０１）。Ｄｅｘ治疗后能显著下调ＴＮＦ－αｍＲＮＡ的表达（６８％,Ｐ〈０．０１）,降低ＡＭ分泌ＴＮＦ－α（Ｐ〈０．０５）及ＩＬ－６水平（Ｐ〈０．０１）。结论 Ｄｅｘ能缓解创伤性ＡＬＩ的发生、发展。其机制与其对ＴＮＦ－α、ＩＬ－６等炎性介质的调节作用密切相关。     

生长因子对间皮细胞增殖及合成细胞外基质的影响

目的　探讨血小板源生长因子ｂｂ（ＰＤＧＦｂｂ） 、转化生长因子β（ＴＧＦβ） 对人腹膜间皮细胞（ＨＰＭＣ）增殖及合成细胞外基质（ＥＣＭ） 的影响。方法　建立ＨＰＭＣ体外培养体系。３ＨＴｄＲ 掺入法测定细胞增殖；放射免疫法检测细胞上清中Ⅲ型前胶原（ ⅢＰＣ） 。结果　ＰＤＧＦｂｂ 促进、但ＴＧＦβ抑制ＨＰＭＣ增殖，均呈剂量依赖性（ Ｐ＜ ０－０１） 。２ 者均能刺激ＨＰＭＣ 产生ⅢＰＣ（ Ｐ＜ ０－０１）。结论　ＣＡＰＤ相关性腹膜炎时腹腔中高浓度的ＰＤＧＦｂｂ 、ＴＧＦβ能调节间皮层的修复和重塑，刺激其合成分泌ＥＣＭ，后者可能是腹膜炎（ＰＩ）反复发生最终导致腹膜硬化的重要机制之一。     

肿瘤坏死因子α对膀胱肿瘤细胞作用的影响因素探讨

目的 探讨肿瘤坏死因子抗膀胱癌细胞作用的影响因素。方法 将肿瘤坏死因子α（ＴＮＦ－α）和不同浓度的丝裂霉素（ＭＣ），顺铂（ＣＤＤＰ）以及干扰素α（ＩＦＮ－α）作用于体外培养的膀胱癌细胞系ＢＩＵ－８７，用噻唑蓝（ＭＴＴ）法测定细胞毒效果。结果 ＴＮＦ－α对ＢＩＵ－８７细胞有明显的浓度依赖性抑制作用，ＭＣ（２．５～５．０ｍｇ／Ｌ）或ＩＦＮ－α（２５００Ｕ－５０００Ｕ／ｍｌ）与ＴＮＦ－α（１００００Ｕ／     

糖尿病纤维连接蛋白血小板聚集功能改变临床分析

为探讨ＮＩＤＤＭ患者Ｆｎ、ＰＡＧ改变与缺血性心脑血管病的关系，分析５３例ＮＩＤＤＭ患者Ｆｕ、ＰＡＧ测定结果，并与５３名健康人比较。患者组Ｆｎ明显高于（Ｐ〈０．０１），前者ＰＡＧ（１）、ＭＡＲ、Ｉ与后者有显著统计学差异（Ｐ〈０．０１）。患者组中并发症组与无并发症组Ｆｎ、Ｉ较对照组有明显统计学差异（Ｐ〈０．０１），并发症组ＰＡＧ（１）、ＭＡＲ明显高于对照组（Ｐ〈０．０５）。ＮＩＤＤＭ患者Ｆｎ降低、ＰＡ     

糖皮质激素对肿瘤坏死因子引起的大鼠中性粒细胞粘附的影响

采用中性粒细胞(PMN)与玻璃珠粘附和PMN与血管内皮细胞(EC)粘附两种模型,以肿瘤坏死因子(TNF),作为PMN的刺激因子,研究糖皮质激素(GC)对TNF引起的大鼠PMN粘附的影响,同时给予糖皮质激素受体(GR)阻断剂RU38486观察GR在粘附中的作用。结果发现,TNF能明显增强大鼠PMN的粘附(P<0.01);Dex不能抑制经TNF预处理的PMN的粘附(P>0.05),但有一定的预防作用;经TNF预处理再同时给予Dex和RU38486的PMN粘附同样明显增强(P<0.01)。     

Tumor necrosis factor-enhanced leukotriene B4 generation and chemotaxis in human neutrophils

In an in vivo study of five normal volunteers infused with endotoxin (20 U/kg of US reference endotoxin lot EC-5), increased neutrophil (PMN) generation of leukotriene B4 and chemotaxis to leukotriene B4 were found concomitantly with elevated plasma tumor necrosis factor (TNF) levels. To clarify the role of TNF in PMN activation, neutrophil responsiveness after in vitro treatment with TNF was examined. Neutrophils from seven normal subjects were incubated with TNF for 30 minutes and tested for chemotaxis to leukotriene B4, formyl-methionyl-leucyl-phenylalanine and zymosan-activated serum, or the calcium ionophore A23187 to assess leukotriene B4 generation. A range of 10(-13) to 10(-9) mol/L of TNF was used for these assays. When 10(-9) mol/L of TNF was used, the amount of leukotriene B4 that was produced was significantly greater than in control cells. The effect of TNF on PMN chemotaxis was uniformly inhibitory for the three stimuli at 10(-10) mol/L compared with untreated cells. At a picomolar range, PMN migration to leukotriene B4, but not to zymosan-activated serum or formyl-methionyl-leucyl-phenylalanine, was significantly increased over that of PMNs not exposed to TNF. This suggests that TNF has a specific facilitatory effect on PMN responsiveness for both leukotriene B4 production and chemotaxis to leukotriene B4 and may be the same signal for this phenomenon in endotoxemic patients.     

CD18 adhesion receptors, tumor necrosis factor, and neutropenia during septic lung injury.

Sequestration of neutrophils (PMNs) in the pulmonary microvasculature and associated neutropenia are characteristic features of experimental models of septic lung injury. The etiology of altered PMN kinetics during septic lung injury is uncertain, but may be partially due to increased adhesiveness of activated PMNs to pulmonary endothelium. This study examines the relationship between the expression of PMN CD18 adhesion receptors, the evolving neutropenia, and plasma tumor necrosis factor (TNF) activity in a porcine model of septic lung injury. Acute lung injury was induced by infusion of live Pseudomonas aeruginosa (5 x 10(8) CFU/ml at 0.3 ml/20 kg/min) for 60 min (Group Ps, n = 6). Control animals (Group C, n = 3) received a 60-min infusion of sterile 0.9% saline. CD18 expression of circulating PMNs was measured by quantitative immunofluorescent flow cytometry. Plasma TNF activity was measured by L929 fibroblast cytolytic assay. Group Ps developed a significant neutropenia by 30 min (14.9 +/- 2.5 vs 23.4 +/- 3.3 x 10(3) cells/microliter at baseline, P less than 0.05, ANOVA) with circulating neutrophils exhibiting significantly increased CD18 expression by 60 min (6.34 +/- 0.72 vs 5.01 +/- 0.52 equivalent soluble fluorescence molecules (ESFM) x 10(3) at baseline, P less than 0.05, ANOVA). Group Ps demonstrated a significant increase in plasma TNF activity by 30 min (2.5 +/- 0.9 vs 0.7 +/- 0.3 U/ml at baseline). There was no significant change in PMN count, PMN CD18 expression, or plasma TNF activity in Group C. In complimentary in vitro studies, porcine PMNs stimulated with recombinant human TNF-alpha (n = 5) demonstrated a time- and dose-dependent increase in CD18 expression.(ABSTRACT TRUNCATED AT 250 WORDS)     

Estrogen‐dependent efficacy of limb ischemic preconditioning in female rats

Our aim was to examine the effects of ischemic preconditioning (IPC) on the local periosteal and systemic inflammatory consequences of hindlimb ischemia‐reperfusion (IR) in Sprague–Dawley rats with chronic estrogen deficiency (13 weeks after ovariectomy, OVX) in the presence and absence of chronic 17beta‐estradiol supplementation (E2, 20 µg kg^?1, 5 days/week for 5 weeks); sham‐operated (non‐OVX) animals served as controls. As assessed by intravital fluorescence microscopy, rolling and the firm adhesion of polymorphonuclear neutrophil leukocytes (PMNs) gave similar results in the Sham + IR and OVX + IR groups in the tibial periosteal microcirculation during the 3‐h reperfusion period after a 60‐min tourniquet ischemia. Postischemic increases in periosteal PMN adhesion and PMN‐derived adhesion molecule CD11b expressions, however, were significantly reduced by IPC (two cycles of 10′/10′) in Sham animals, but not in OVX animals; neither plasma free radical levels (as measured by chemiluminescence), nor TNF‐alpha release was affected by IPC. E2 supplementation in OVX animals restored the IPC‐related microcirculatory integrity and PMN‐derived CD11b levels, and TNF‐alpha and free radical levels were reduced by IPC only with E2. An enhanced estrogen receptor beta expression could also be demonstrated after E2 in the periosteum. Overall, the beneficial periosteal microcirculatory effects of limb IPC are lost in chronic estrogen deficiency, but they can be restored by E2 supplementation. This suggests that the presence of endogenous estrogen is a necessary facilitating factor of the anti‐inflammatory protection provided by limb IPC in females. The IPC‐independent effects of E2 on inflammatory reactions should also be taken into account in this model. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:97–105, 2018.

     

Cytokines enhance the production of a chemotactic factor for polymorphonuclear leukocytes by rat renal glomerular epithelioid cells

Human recombinant interleukin-1 beta (IL-1) and tumor necrosis factor-alpha (TNF) dose-dependently enhanced the production of PMN chemotactic factor, which was trypsin-sensitive and heat-stable, by the epithelioid cells of rat renal glomeruli. Dexamethasone dose-dependently suppressed the chemoattractant production. Molecular weight and isoelectric point of this factor were 10 kDa and over 10, respectively. These results suggest that PMN migration to glomeruli occurs without participation of complement system when TNF or/and IL-1 are produced by activated cells in glomeruli.     

烧伤后白细胞与内皮细胞粘附分子介导作用的研究

为研究白细胞粘附依赖于细胞粘附分子的介导作用,作者通过离体实验观察分析了烧伤血清对外周血中性粒细胞（ＰＭＮ）ＣＤ１１ｂ／ＣＤ１８表达的影响及ＣＤ１１ｂ／ＣＤ１８在烧伤后ＰＭＮ粘附中的介导作用。结果表明：（１）烧伤血清使ＰＭＮＣＤ１１ｂ／ＣＤ１８分子表达和ＰＭＮ与肺微血管内皮细胞（ＰＭＥＣ）的粘附率增高。（２）ＣＤ１１ｂ／ＣＤ１８单抗不仅能使正常ＰＭＮ与ＰＭＥＣ的粘附率下降５０％,也使烧伤血清激活的ＰＭＮ与ＰＭＥＣ粘附率下降至烧伤血清激活前水平。我们认为,严重烧伤能使外周血ＰＭＮＣＤ１１ｂ／ＣＤ１８表达明显增高,ＣＤ１１ｂ／ＣＤ１８不但介导基础状态下的ＰＭＮ粘附,更是介导烧伤后引起的大量ＰＭＮ与ＰＭＥＣ粘附的主要分子。     

烟雾吸入伤血清对中性粒细胞粘附及迁移作用的体外研究

目的 通过鼠烟雾吸入伤血清体外刺激培养内皮细胞和中性粒细胞（ＰＭＮ），了解ＰＭＮ粘会与迁移变化，以及抗ＣＤ１１ａ，抗ＣＤ１１ｂ和抗ＩＣＡＭ－１对ＰＭＮ粘附与迁移的影响，探讨粘附因子在烟雾吸入性损伤中的作用。方法 在体外实验中用荧光标记ＰＭＮ，测定ＰＭＮ与烟雾吸入性损伤血清刺激的内皮细胞的粘附率，在５μｍ滤膜上培养仙皮细胞测定ＰＭＮ迁移量，并采用抗体阻断技术，测定了粘附因子ＣＤ１１ａ，ＣＤ１１ｂ和Ｉ     

烧伤后粘附分子介导中性粒细胞粘附对肺血管通透性的影响

目的观察比较烧伤血清及烧伤后中性白细胞（ＰＭＮ）对肺血管通透性的影响,分析ＰＭＮ粘附及粘附分子ＣＤ１１ｂ／ＣＤ１８在该影响中的介导作用。方法应用离体肺灌流技术,通过肺重量增加（ＬＷＧ）、液体滤过系数（Ｋｆ）和通透性表面积乘积（ＰＳ）分别观察肺水肿程度、肺血管对小分子物质和大分子物质的通透性。结果烧伤血清能使ＬＷＧ、Ｋｆ和ＰＳ明显增加,以Ｋｆ增加最为明显;烧伤后ＰＭＮ也能使Ｋｆ和ＰＳ增加,以ＰＳ增加为明显;用单抗封闭烧伤后ＰＭＮ膜上ＣＤ１１ｂ／ＣＤ１８后,ＰＭＮ在肺血管内的滞留减少,Ｋｆ和ＰＳ值增加被抑制,并以ＰＳ改变为显著。结论（１）被激活的ＰＭＮ释放的介质类物质如氧自由基、蛋白酶等物质对肺微血管内皮细胞（ＰＭＥＣ）的损伤作用部分依赖于ＰＭＮ与内皮细胞的粘附。（２）烧伤后被激活的ＰＭＮ释放的介质类物质主要介导肺血管对小分子物质通透性的影响。烧伤后ＰＭＮ与肺微血管内皮细胞（ＰＭＥＣ）的粘附除了使介质类物质的作用放大外,还介导肺血管对大分子物质通透作用。（３）ＰＭＮ膜上ＣＤ１１ｂ／ＣＤ１８分子可能通过与ＰＭＥＣ细胞间粘附分子的结合,本身具有对内皮细胞的生物学调控作用     

Alterations in immunocyte tumor necrosis factor receptor and apoptosis in patients with congestive heart failure

OBJECTIVE: To examine systemic immune cell proinflammatory receptor expression and apoptosis in patients with congestive heart failure (CHF). SUMMARY BACKGROUND DATA: Prior studies have demonstrated that CHF is associated with a chronic myocardial inflammatory state, including increased plasma proinflammatory cytokine and soluble receptor expression. By contrast, it has also been shown that tumor necrosis factor (TNF) receptor protein expression is decreased in the failing myocardium. However, no studies to date have examined systemic immune cell proinflammatory receptor expression or function as disease markers in patients with heart failure. METHODS: Twenty-nine patients were studied prospectively over an 8-month period at a single institution. One group (n = 16) had a history of clinical symptoms of CHF and moderate to severe left ventricular dysfunction. The second group (n = 13) consisted of patients who had coronary artery disease without symptoms of CHF and documented preservation of left ventricular function. Blood samples were analyzed for polymorphonuclear cell (PMN) and monocyte TNF and CD95 membrane-associated receptor expression, spontaneous and CD95 (Fas)-mediated PMN apoptosis, and plasma cytokine and soluble TNF receptor levels. Isolated PMNs were incubated for 6 hours with or without CH 11, a CD95 agonist. Propidium iodide/RNAase staining and flow cytometry was used to assess apoptosis, defined as PMNs expressing hypodiploid DNA (<2 n DNA). Membrane-associated TNF receptor and CD95 were also measured by flow cytometry. Plasma levels of TNF, interleukin (IL)-6, IL-10, and soluble TNF receptors 1 and 2 were quantified using enzyme-linked immunosorbent assay. RESULTS: Compared to patients without CHF, circulating PMN and monocyte TNF receptor levels were significantly decreased in patients with CHF. By contrast, PMN and monocyte CD95 expression was not significantly changed in patients with CHF versus those without CHF. Patients with CHF had a 60% decrease in spontaneous PMN apoptosis compared to patients without CHF, whereas no significant difference in CD95-mediated apoptosis was observed between the two groups. Pearson-product movement correlation of monocyte TNF receptor expression and spontaneous PMN apoptosis rates versus patients' ejection fraction was performed and was statistically significant. Plasma levels of soluble TNF receptor 2 (p75) were elevated in CHF patients versus patients without CHF, while there was no significant difference in soluble TNF receptor 1 (p55), TNF, IL-6, and IL-10 between the two groups. CONCLUSIONS: These data demonstrate a systemic alteration in immune cell phenotype and apoptosis in patients with CHF. These findings provide support for the concept that inflammatory mediators either contribute to myocardial dysfunction or are elaborated systemically by left ventricular compromise. This present study suggests that immune cell TNF receptor expression and diminished PMN apoptosis may serve as biologic markers of myocardial failure.