姜黄素通过IL-6/STAT3信号通路调控Th17/Treg平衡治疗溃疡性结肠炎

目的:观察姜黄素对溃疡性结肠炎(UC)模型小鼠的疗效及对IL-6/STAT3信号通路中相关分子表达和结肠组织中Treg、Th17细胞水平的影响,探究姜黄素是否通过IL-6/STAT3信号通路调控Th17/Treg平衡以有效治疗UC。方法:采用葡聚糖硫酸钠(DSS)制备UC小鼠模型,将50只健康雌性BALB/c小鼠随机分为5组:正常对照组,UC模型组及姜黄素低、中、高剂量组。HE染色观察小鼠结肠黏膜组织损伤程度并评分,Western blot检测结肠组织中IL-6、STAT3及p-STAT3的蛋白水平,流式细胞术分析结肠组织中Treg和Th17细胞的水平,Western blot检测结肠组织中Treg转录因子FOXP3和Th17转录因子RORγt的表达情况。结果:UC小鼠结肠部位溃疡形成,镜下可见炎症和充血等病理改变,姜黄素中剂量组的黏膜损伤基本恢复正常,只有少量炎性细胞。与正常组相比,UC小鼠结肠组织IL-6、STAT3和p-STAT3的蛋白水平均显著增高;与模型组相比,姜黄素中剂量组IL-6、STAT3和p-STAT3的蛋白水平均明显降低,接近正常组,并且中剂量姜黄素能显著降低Th17细胞的水平和转录因子RORγt的表达,而显著升高Treg细胞的水平和转录因子FOXP3的表达。姜黄素低剂量组和高剂量组对上述指标均无明显影响。结论:姜黄素可以有效治疗DSS诱导的小鼠UC,其作用机制可能是通过IL-6/STAT3信号通路来调控Th17/Treg平衡。     

金纳多对小鼠急性实验性结肠炎的保护作用及可能机制

目的观察金纳多对小鼠急性实验性结肠炎的保护机制并对比分析其与柳氮磺胺吡啶(SASP)的疗效。方法用3%DSS建立小鼠急性实验性结肠炎动物模型。小鼠随机分为4组:正常组、造模组、金纳多治疗组、柳氮磺胺吡啶(SASP)治疗组。记录各组小鼠一般情况(饮食、便血、体重)、DAI评分、肠管长度和肠组织学损伤评分;通过免疫组化、Real-time PCR和Western blot检测各组小鼠肠组织中炎症因子(gp130,STAT3, p-STAT3, ROR-γt,IL-6, IL-17, IL-23)的表达。结果金纳多可改善小鼠一般情况(饮食、便血、体重),与造模组比较,可有效降低小鼠DAI评分、组织学损伤评分、炎症因子(gp130, STAT3, p-STAT3, ROR-γt,IL-6, IL-17, IL-23)的表达水平(P0.01)。与SASP治疗组比较,金纳多治疗组小鼠DAI评分、组织学损伤评分、炎症因子(gp130, STAT3, p-STAT3, ROR-γt,IL-6, IL-17, IL-23)的表达均高于SASP治疗组(P0.05)。结论金纳多对小鼠急性实验性结肠炎的保护作用与抑制IL-6/STAT3通路和IL-23/IL-17轴相关,但疗效弱于柳氮磺胺吡啶。     

基于IL-6/JAK2/STAT3信号通路探讨灰树花提取物对溃疡性结肠炎大鼠结肠组织炎症反应的影响

目的：探讨灰树花提取物通过调控白细胞介素6(IL-6)/蛋白酪氨酸激酶2(JAK2)/信号转导和激活因子3(STAT3)信号通路,对溃疡性结肠炎（UC）大鼠结肠组织炎症反应的影响及作用机制。方法：将40只SD大鼠随机分为空白对照组、UC模型组、灰树花治疗组、西药治疗组、联合治疗组,每组8只。采用3%DSS自由饮用法7 d建立UC大鼠模型后,治疗组分别灌胃灰树花提取物10 mg/（kg·d）、柳氮磺吡啶0.3 g/（kg·d）及等量两种药物,连续14 d。实验期间观察大鼠一般状态,计算疾病活动指数（DAI）评分;采用HE染色法观察大鼠结肠组织病理改变;Western blot检测大鼠结肠组织IL-6、JAK2、STAT3、pSTAT3蛋白表达水平;ELISA检测大鼠血清中IL-6含量;免疫组化法测定大鼠结肠组织中IL-6R、MPO蛋白含量及表达情况。结果：与空白对照组比较,UC模型组大鼠一般状态欠佳,DAI评分升高,HE染色可见明显的组织黏膜损伤与炎症细胞浸润;结肠组织IL-6、JAK2、STAT3、p-STAT3蛋白表达水平及IL-6R、MPO含量均显著升高（P<0.01）;血清...     

IL-18在脓毒症肺损伤发生发展过程中的作用及机制研究北大核心CSCD

目的:研究IL-18在脓毒症肺损伤发展过程中的作用及调控机制。方法:成年雄性野生型C57BL/6(WT)和IL-18基因敲除(IL-18-/-)小鼠分为野生型小鼠对照组(WT组)、脂多糖(LPS)处理的野生型小鼠组(WT LPS组)、IL-18基因敲除的小鼠对照组(IL-18-/-组)、LPS处理的IL-18基因敲除小鼠组(IL-18-/-LPS组)。腹腔注射LPS(15 mg/kg)建立小鼠脓毒症模型,对照组注射等量生理盐水。观察各组小鼠72 h生存率并处死小鼠,HE染色观察肺部病理组织变化,RT-PCR及Western blot检测各组小鼠肺组织IL-18 mRNA及蛋白表达,免疫荧光检测肺组织IL-18表达及定位,TUNEL染色检测肺组织细胞凋亡,流式细胞术检测肺组织Treg/Th17比例,ELISA检测炎症因子TNF-α、IL-17A、TGF-1β、IL-10表达。Western blot检测各组小鼠肺组织RORγt、FoxP3蛋白表达及STAT3磷酸化水平。结果:LPS诱导后,小鼠肺组织高表达IL-18。与WT LPS组相比,IL-18-/-LPS组小鼠生存率显著提高,肺组织病理损伤减轻,凋亡细胞显著减少,Treg/Th17比例提高,抑炎因子TGF-1β、IL-10表达增加,而促炎因子TNF-α、IL-17A表达明显减少,肺组织RORγt蛋白表达增加,而STAT3磷酸化水平降低。结论:IL-18可通过上调STAT3磷酸化水平,促进Treg/Th17免疫失衡及炎症因子分泌,从而加剧脓毒症急性肺损伤。     

温补脾肾法对脾肾阳虚型UC大鼠JAK2/STAT3-SOCS6信号通路相关蛋白及基因表达的影响北大核心CSCD

目的:观察以温补脾肾法为指导的理中汤合四神丸对脾肾阳虚型溃疡性结肠炎(UC)大鼠模型病变结肠组织中JAK2、STAT3、p-STAT3、SOCS6蛋白及基因表达的影响。方法:采用病证结合造模方法复制脾肾阳虚型UC模型,成模大鼠随机分为模型组、柳氮磺砒啶组(SASP组)、理中汤合四神丸高、中、低剂量组,16只/组,同时设16只大鼠作为空白组。空白组与模型组给予蒸馏水灌胃,各治疗组给予对应的药物及剂量灌胃治疗,连续21 d。HE染色观察大鼠结肠黏膜组织病理变化;免疫组化法检测大鼠结肠黏膜组织中JAK2、STAT3、p-STAT3、SOCS6蛋白表达情况;RT-qPCR检测大鼠JAK2、STAT3、SOCS6mRNA表达情况;ELISA检测大鼠组织和血清中TGF-β1、ICAM-1含量。结果:与空白组相比,模型组大鼠结肠黏膜组织JAK2、STAT3、p-STAT3蛋白表达显著升高(P<0.01),JAK2、STAT3、p-STAT3蛋白着色深,表达呈强阳性,SOCS6蛋白表达显著降低(P<0.01),SOCS6蛋白着色浅,表达呈弱阳性。JAK2、STAT3 mRNA表达显著升高,SOCS6 mRNA表达显著降低(P<0.01),组织和血清中TGF-β1含量显著降低(P<0.01),ICAM-1含量显著升高(P<0.01);与模型组相比,中药高、中剂量组及SASP组JAK2、STAT3、p-STAT3蛋白表达显著降低(P<0.01),JAK2、STAT3、p-STAT3蛋白着色浅,表达呈弱阳性,SOCS6蛋白表达显著升高(P<0.01),SOCS6蛋白着色深,表达呈强阳性。JAK2、STAT3 mRNA表达显著降低(P<0.01),SOCS6 mRNA表达显著升高(P<0.01),组织和血清中TGF-β1含量显著升高(P<0.01),ICAM-1含量显著降低(P<0.01)。结论:以温补脾肾法为指导的理中汤合四神丸可能通过抑制JAK2/STAT3-SOCS6信号通路的激活,调节免疫系统平衡,修复受损结肠黏膜,达到治疗UC的目的。     

养血解毒汤对咪喹莫特诱导的STAT3转基因小鼠银屑病样皮损的干预作用

目的:观察养血解毒汤对咪喹莫特诱导的STAT3转基因小鼠银屑病样皮损的干预作用。方法:STAT3转基因小鼠24只,随机分为正常对照组、模型组、养血解毒汤组和甲氨蝶呤组,每组6只,采用外用咪喹莫特诱导皮肤银屑病样模型。分别观察皮损面积和疾病严重程度(psoriasis area and severity index,PASI),光镜下观察皮损组织形态学变化和表皮层厚度;采用皮肤水分油分测试笔检测小鼠背部皮肤水分和油分含量;免疫组织化学法检测皮损中增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)和T淋巴细胞表面标志CD3表达;采用实时荧光定量PCR技术检测皮损Th17类细胞因子(IL-17A、IL-17C、IL-22和RORγt)mRNA表达;采用Western blot检测皮损STAT3通路蛋白(STAT3、p-STAT3、JAK3、p-JAK3和SOCS3)的表达水平。结果:(1)养血解毒汤组PASI评分明显低于模型组,HE染色观察发现养血解毒汤组皮损表皮增生较模型组少,角化不全的细胞明显减少,皮肤厚度降低(P0.01);(2)养血解毒汤组皮损油分和水分含量均明显高于模型组(P0.05);(3)养血解毒汤组PCNA阳性表达远低于模型组(P0.01);(4)养血解毒汤组脾脏重量低于模型组(P0.05);(5)养血解毒汤组Th17相关因子(IL-17A、IL-17C、IL-22和RORγt)表达水平均低于模型组,此外,p-STAT3和p-JAK3表达量亦低于模型组,而SOCS3的表达高于模型组。结论:养血解毒汤可能通过抑制STAT3的磷酸化,减少Th17相关因子(IL-17A、IL-17C、IL-22和RORγt)的分泌,由此改善咪喹莫特诱导的STAT3转基因小鼠银屑病样皮损改变。     

地塞米松对哮喘小鼠肺组织中转录因子RORγt mRNA表达的干预作用

目的:初步探讨地塞米松对哮喘小鼠模型肺组织中Th17细胞转录因子维甲酸相关孤儿受体γt(RORγt)mRNA表达的影响.方法:采用卵清蛋白致敏方法建立支气管哮喘小鼠模型,BALB/c小鼠30只随机分为正常对照组、哮喘组、地塞米松治疗组各10只.采用酶联免疫吸附试验(ELISA)技术检测小鼠肺泡灌洗液(BALF)、血清中白细胞介素-17(IL-17)水平;无创肺功能体描仪检测小鼠气道反应性;HE染色评价各组小鼠气道炎症程度;逆转录聚合酶链反应(RT-PCR)方法检测肺组织IL-17、RORγt mRNA表达水平.结果:哮喘组小鼠肺组织RORγt mRNA、IL-17水平高于对照组(P<0.01),地塞米松治疗组RORγt mRNA、IL-17水平低于哮喘组(P<0.05).结论:地塞米松可抑制哮喘小鼠肺组织RORγt表达,阻止Th17的分化,从而减少IL-17的分泌,减轻哮喘气道炎症反应.     

蚓激酶对哮喘小鼠免疫相关转录因子T-bet、GATA-3、RORγt、Foxp3的影响北大核心CSCD

目的:观察蚓激酶对哮喘小鼠炎症因子IL-6、TGF-β、IL-1β、TNF-α水平和免疫相关转录因子T-bet、GATA-3、RORγt、Foxp3 mRNA及蛋白表达的影响。方法:40只BALB/c雄性小鼠随机分为4组:空白对照组(空白组)、哮喘模型组(模型组)、地塞米松组(地米组)和蚓激酶组,每组10只。第1、14天模型组、地米组和蚓激酶组腹腔注射OVA+Al(OH)3致敏,21~27 d雾化吸入5%OVA建立哮喘模型,每次雾化激发1 h后,地米组、蚓激酶组分别灌胃给予地塞米松溶液、蚓激酶溶液治疗7 d,空白组、模型组灌胃等量生理盐水。末次给药24 h后处死取材,HE染色观察肺组织病理学改变,ELISA检测支气管肺泡灌洗液(BALF)中IL-6、IL-1β、TGF-β、TNF-α水平,RT-PCR检测肺组织T-bet、GATA-3、RORγt、Foxp3 mRNA水平,Western blot检测肺组织T-bet、GATA-3、RORγt、Foxp3蛋白表达。结果:肺组织病理观察显示,模型组小鼠管腔及周围炎症细胞浸润明显,黏膜水肿增厚,地米组和蚓激酶组小鼠肺部炎症浸润及黏膜增厚水肿均较模型组好转。与空白组相比,模型组IL-6、TGF-β、IL-1β、TNF-α水平显著升高,GATA-3、RORγt mRNA及蛋白表达增加(P<0.01),T-bet、Foxp3 mRNA及蛋白表达降低(P<0.01)。与模型组相比,地米组和蚓激酶组IL-6、TGF-β、IL-1β、TNF-α水平降低(P<0.05,P<0.01),T-bet、Foxp3 mRNA表达升高(P<0.01),GATA-3、RORγt mRNA及蛋白表达降低(P<0.01)。结论:蚓激酶可降低炎症因子IL-6、TGF-β、IL-1β及TNF-α水平,抑制GATA-3、RORγt mRNA及蛋白表达,诱导T-bet、Foxp3 mRNA及其蛋白表达,且可能通过降低相关前炎症因子水平调节免疫细胞转录因子表达。     

哮喘小鼠肺组织中转录因子RORγt的表达与气道炎症的关系

目的:探讨Th17细胞转录因子RORγt在支气管哮喘小鼠肺组织中的表达及其与哮喘气道炎症的关系.方法:采用卵清蛋白(OVA)致敏方法建立支气管哮喘小鼠模型;BALB/c小鼠30只随机分为对照组、哮喘组、地塞米松治疗组各10只.采用酶联免疫吸附试验(ELISA)方法检测小鼠肺泡灌洗液(BALF)、血清中白细胞介素17(IL-17)水平;HE染色评价各组小鼠气道炎症情况;逆转录聚合酶链反应(RT-PCR)方法检测肺组织IL-17、RORγt mRNA表达水平;免疫印记(Western blot)方法检测肺组织RORγt蛋白表达水平.结果:哮喘组小鼠肺组织RORγt mRNA、蛋白水平及IL-17水平均明显高于对照组和地塞米松治疗组(P＜0.05),RORγt蛋白表达量与嗜酸性粒细胞数、淋巴细胞数、中性粒细胞数、BALF、外周血中IL-17含量、肺组织IL-17 mRNA表达量均呈正相关关系(r＝0.789、0.795、0.902、0.669、0.806、0.883,P值均＜0.01).结论:RORγt在支气管哮喘小鼠肺组织呈高表达,其表达水平与气道炎症密切相关,参与了哮喘气道炎症的发生过程.     

The effect of anode curvature on radiographic heel effect

Previous computer modeling of radiographic heel effect has shown that a simple model can give an accurate representation of heel effect for flat anode surfaces. This paper details the extension of this model to allow curved anode surfaces. A fixed-anode x-ray tube with a demountable anode was constructed and used to test the predictions of the computational model at 25 kVp using stainless-steel anodes. The computer model was also used to estimate the magnitude of the heel effect for a tungsten anode at 60 kVp. This work allows determination of the effect of anode curvature, which is an inherent feature of certain recently proposed tube designs for scanning slit radiography.     

Corticosteroid effect on immunoglobulins

The corticosteroid (prednisone) effect on serum immunoglobulins in 9 atopic asthmatic patients who required corticosteroids for the control of asthma was evaluated. Serum immunoglobulins were determined before corticosteroids were administered, an average of 15 days while on corticosteroids, and again an average of 22 days after corticosteroids were discontinued. While on corticosteroids (averaging 16.8 mg prednisone daily) for 15 days, mean serum IgG was significantly decreased (?22%, p ≤ 0.01), mean serum IgA tended to be decreased (?10%), and mean serum IgM was essentially unchanged. Serum IgE was initially significantly increased (p < 0.01) when compared to levels of other serum immunoglobulins (IgG,A,M). An average of 22 days after corticosteroids were discontinued, mean serum IgG was still significantly decreased (p < 0.05), and mean serum IgA again tended to be decreased. Serum IgM remained unchanged and mean serum IgE now was significantly decreased (p < 0.01). Corticosteroids appear to have a significant effect on levels of some serum immunoglobulins.     

Lifestyle effect on total IgE

Recent studies have demonstrated an increasing number of cases of allergy in economically developed societies. Many of these allergic reactions are mediated by IgE antibodies whose levels are under genetic control. However, this large increase of allergy prevalence cannot be explained by genetic mutations; comprehensive lifestyles may also play roles in determining total IgE levels. Total IgE levels were determined from serum samples taken from 706 hard metal workers who responded to a questionnaire including 17 items related to physical and mental health practices. Five factors tending to normalize IgE upward and three tending to normalize IgE downward when we keep good lifestyles were extracted using MANOVA analysis. We therefore constructed a Health Practice Index in Allergic reactions (HPIA); eight physical and mental health practices were significantly (P less than 0.05) associated with cumulative elevations or suppressions of total IgE levels when sex, age and environmental factors were controlled for. Multivariate analyses demonstrated a dramatic synergism (Relative Risk greater than 2.0, P less than 0.05) of unhealthy lifestyles determining a bipolar high risk for elevated or suppressed IgE levels which in turn may result in allergic diseases or immuno-suppression. Maintenance of healthy lifestyles may prove beneficial for reducing the prevalence of allergic disorders.     

Additive effect of an immunomodulator on the therapeutic effect of antibiotics

The effect of an immunomodulator, the Immunoferon (IF), on the course of experimental infections with Staphylococcus aureus and on cellular immune reactions (foot pad swelling test) was examined. The substance is a glycophosphopeptide adsorbed on sulfurized calcium phosphate. The IF improves clearly the therapeutic effect of penicillin, but has no significant effect when given alone. The substance neutralizes the negative effect of hydrocortisone on the therapeutic effectivity of penicillin. The cellular immune reaction is reduced by very small doses of IF, but is enhanced progressively by higher doses. The effect of hydrocortisone is abolished in this respect.     

Clavulanic acid. Its effect on Staphylococcus aureus other than its inhibitory effect on beta-lactamases

The influence of clavulanic acid (CA) on growth kinetics of Staph. aureus strains with MIC values (minimal inhibitory concentration) of penicillin (Pc) between 0,06 and 1 024 IU/ml was examined. Furthermore, it was tried to select mutants of the parent strains with increased resistance to CA and, in the affirmative case, to find out a relationship between Pc and CA resistance. The experiments showed that CA in subinhibitory concentrations induced a considerable increase of the latent phase of Pc sensitive strains but less of the Pc resistant ones. CA delayed also the generation time but this effect was apparently independent of Pc resistance. Successive subculturing of strains in gradually increasing CA concentrations resulted in an increase of CA resistance by a factor of 4 to 8 compared with that of the parent strains. In parallel, the Pc resistance increased by a factor of 16 to 64, however only in case of strains originally sensitive to Pc. A synergistic or additive effect of CA and Pc evaluated by the index of the fractional inhibitory concentration (IFIC) was not only shown in Pc resistant but also in Pc sensitive strains. This effect was equally demonstrable in strains with increased CA resistance. In this case, however, the MIC of Pc in the presence of CA was increased by a factor of 32 to 64 in originally Pc sensitive strains and by a factor of 2 to 4 in the originally Pc resistant ones. These results indicate a possible selection of CA resistant strains with an increased Pc resistance.     

The effect of sound intensity on the audiotactile crossmodal dynamic capture effect

We investigated the effect of varying sound intensity on the audiotactile crossmodal dynamic capture effect. Participants had to discriminate the direction of a target stream (tactile, Experiment 1; auditory, Experiment 2) while trying to ignore the direction of a distractor stream presented in a different modality (auditory, Experiment 1; tactile, Experiment 2). The distractor streams could either be spatiotemporally congruent or incongruent with respect to the target stream. In half of the trials, the participants were presented with auditory stimuli at 75 dB(A) while in the other half of the trials they were presented with auditory stimuli at 82 dB(A). Participants’ performance on both tasks was significantly affected by the intensity of the sounds. Namely, the crossmodal capture of tactile motion by audition was stronger with the more intense (vs. less intense) auditory distractors (Experiment 1), whereas the capture effect exerted by the tactile distractors was stronger for less intense (than for more intense) auditory targets (Experiment 2). The crossmodal dynamic capture was larger in Experiment 1 than in Experiment 2, with a stronger congruency effect when the target streams were presented in the tactile (vs. auditory) modality. Two explanations are put forward to account for these results: an attentional biasing toward the more intense auditory stimuli, and a modulation induced by the relative perceptual weight of, respectively, the auditory and the tactile signals.

Investigating the effect of a transparent barrier on the crossmodal congruency effect

A transparent barrier, such as a window, protects us from approaching objects (such as flies), although we can still see the objects coming toward us through the occluder. In the present study, we examined whether this potential dissociation between tactile and visual experience introduced by the presence of a transparent barrier also affects visual-tactile interactions in normal participants, as indexed by performance in the crossmodal congruency task. Participants discriminated the elevation of vibrotactile target stimuli (upper vs lower) presented to the left or right hand while trying to ignore visual distractor lights that could independently be presented from upper or lower locations on either the same or the opposite side. A transparent occluder was placed between the vibrotactile targets and the visual stimuli (the barrier occluded the vibrotactile targets in Experiment 1 and the visual distractors in Experiment 2). Vibrotactile elevation discrimination performance was slower and less accurate when the distractor lights were presented from incongruent locations relative to the target (e.g., lower tactile target with upper distractor light). However, there were no significant differences between performance with and without the transparent occluder present. This pattern of results was replicated in Experiment 3 under conditions where the participants were periodically required to reach around the transparent occluder in order to press buttons placed near to the visual distractors. Taken together, these results support recent neuropsychological evidence [Farne et al. (2003) Int J Psychophysiol 50:51-61] suggesting that visual-tactile interactions in peripersonal space are not necessarily modulated by conscious awareness of the impossibility of our hand being touched by the visual stimuli.