IL-12单克隆抗体对克罗恩病模型小鼠Th1/Th2免疫平衡的调节作用

目的研究白细胞介素-12(IL-12)单克隆抗体对克罗恩病模型小鼠辅助性T细胞1/辅助性T细胞2(Th1/Th2)免疫平衡的调节作用。方法选择BALB/c健康小鼠30只,将30只小鼠分为正常对照组(10只)和实验小鼠(20只),实验组小鼠做克罗恩病模型,建模成功后将实验组小鼠分为克罗恩病组和IL-12单克隆抗体组各10只,IL-12单抗组小鼠腹腔注射IL-12单抗(25 mg/kg),正常对照组和克罗恩病组小鼠腹腔注射0.2 ml的生理盐水。处理4周后,对三组小鼠疾病活动程度、结肠重量、结肠长度进行评价,使用酶联免疫吸附试验法检测三组小鼠结肠组织中Th1型细胞因子干扰素-γ(IFN-γ)、肿瘤坏死因子-α(TNF-α)、IL-1β、IL-8和Th2型细胞因子IL-4、IL-10及IL-12、IL-23表达水平。结果克罗恩病组小鼠疾病活动程度评分、结肠组织中Th1型细胞因子IFN-γ、TNF-α、IL-1β、IL-8、IL-12、IL-23分别为(4.12±0.23)分、(563.49±21.36)ng/ml、(120.69±6.69)ng/L、(43.73±3.58)ng/ml、(179.68±4.26)ng/ml、(148.69±6.58)ng/L、(762.58±26.58)pg/ml高于IL-12单克隆抗体组(1.62±0.17)分、(472.59±15.64)ng/ml、(106.25±3.27)ng/L、(37.62±2.16)ng/ml、(163.57±3.12)ng/ml、(98.67±5.12)ng/L、(412.68±16.98)pg/ml)和对照组,而结肠重量/长度为(42.36±3.16)mg/cm大于IL-12单克隆抗体组(33.59±2.07)mg/cm)和对照组小鼠;结肠组织中Th2型细胞因子IL-4、IL-10为(55.27±1.62)pg/ml、(90.27±3.68)pg/ml低于IL-12单克隆抗体组(61.27±2.21)pg/ml、(98.67±1.68)pg/ml)和对照组(P<0.05)。结论IL-12单克隆抗体通过改善Th1型细胞因子、Th2型细胞因子的异常表达,对Th1/Th2免疫平衡进行调节,进而抑制克罗恩病的炎症反应,起到治疗的效果。     

Vanin对胰岛NIT细胞保护机制的探讨

目的探讨泛酰巯基乙胺酶Vanin对胰岛NIT细胞的保护作用及机制。方法培养胰岛B细胞株NIT细胞,以5ng／ml IFN-1,50pg／ml IL-1β,10ng／ml Vanin处理细胞,分为IFN-γ＋IL-1β[3＋Vanin组、IFN-γ＋IL-1β组、Vanin组、对照组（DMEM）。4组干预因素分别作用于对数生长期细胞24h后采用MTF法检测各组NIT细胞的增殖抑制率,化学发光法检测上清液中胰岛素（Ins）水平,硝酸还原酶法检测上清液中一氧化氮（N0）水平。结果经IFN-γ＋IL-1β处理的NIT-1细胞增殖受抑制,抑制率为60．11％;予Vanin预处理组细胞增殖抑制率为36．98％,较IFN-γ＋IL-1β处理组降低（P〈O．01）。经IFN-γ＋IL-1β破坏的NIT-1细胞分泌胰岛素较其他3组减少（P〈O．05,P〈0．01）,NO产生较其他3组增多（P〈0．05,P〈0．01）。而接受Vanin预处理组再予IL-1β＋IFN-γ破坏的NIT-1细胞与IL-1β＋IFN-γ叫组比较,分泌胰岛素增加（P〈0．05）、NO水平降低（P〈0．05）。结论Vanin可减轻IFN-γ＋IL-1β对NIT细胞的损伤作用,保护NIT细胞的胰岛素分泌功能。     

肺结核患者血清IL-6、IL-12、IL-23、TNF-α、IFN-γ表达水平的临床意义分析

目的探讨肺结核患者血清中巨噬细胞相关细胞因子IL-6、IL-12、IL-23、TNF-α、IFN-γ的变化及其临床意义。方法采用酶联免疫吸附法(ELISA)检测30例活动性肺结核患者(患者);30例结核分枝杆菌感染者(感染者);30例健康人血清中的IL-6、IL-12、IL-23、TNF-α、IFN-γ分泌水平。结果 IL-6在健康人、患者者和感染组血清中的分泌水平分别为:0.43±0.03 pg/ml,0.86±0.04 pg/ml,0.46±0.02 pg/ml;IL-12在健康人、患者者和感染组血清中水平分别为:9.43±0.22 pg/ml,14.96±0.58 pg/ml,9.58±0.20 pg/ml;IL-23在健康人、患者者和感染组血清中水平分别为:269.58±28.58pg/ml,336.58±30.61 pg/ml;332.03±25.71 pg/ml;IFN-γ在健康人、患者者和感染组血清中水平分别为:90.80±2.05 pg/ml,116.15±4.96 pg/ml,114.29±3.16 pg/ml;TNF-α在健康人、患者者和感染组血清中水平分别为:13.87±3.28 pg/ml,14.06±2.14 pg/ml,13.43±3.12 pg/ml。统计分析发现:IL-6、IL-12、IL-23、IFN-γ在患者血清中的分泌水平显著高于健康对照组(p<0.05);IL-23与INF-γ在感染者血清中的分泌水平显著高于健康对照组(p<0.05);IL-6与IL-12在患者血清中的分泌水平显著感染者组(p<0.05);其余细胞因子在3类人群血清中分泌水平差别无统计学意义。结论血清中巨噬细胞相关细胞因子IL-6、IL-12、IL-23、IFN-γ在肺结核发病中具有重要意义,具有成为肺结核临床诊断依据可能。     

孕妇外周血基质金属蛋白酶-9、白介素-6水平联合B超测定宫颈长度预测早产的研究

目的探讨孕妇外周血基质金属蛋白酶一9（MMP-9）、白介素-6（IL-6）及宫颈长度对早产预测的应用价值。方法选择本院正常妊娠组孕妇23例,先兆早产孕妇39例,按是否继续妊娠为2个亚组,早产分娩组18例,继续妊娠组21例,分别采集各组静脉血,分离血浆应用ELISA方法测定MMP-9及IL-6水平,同时采用B超测定宫颈长度,预测孕妇早产发生情况。结果（1）早产分娩组孕妇MMP-9及IL-6水平显著高于正常妊娠组及继续妊娠组[（47．24±6．41）ng／ml,（523．24±62．35）pg／mlvs（20．39±7．23）ng／ml,（219．29±80．23）pg／ml,（32．18±5．16）ng／ml,（406．18．4-71．52）pg／ml],其差异均有统计学意义（P〈0．01）,继续妊娠组血浆MMP-9及IL-6水平显著高于正常妊娠组[（32．18±5．16）ng／ml,（406．18±71．52）pg／mlvs（20．39±7．32）ng／ml,（219．29±80．23）pg／ml],其差异均有统计学意义（P〈0．01）;（2）早产分娩组宫颈长度显著低于正常妊娠组及继续妊娠组[（15．62±2．31）mmVS（31．05±1．48）mm,（26．14±3．12）mm,P〈0．01],继续妊娠组孕妇宫颈长度稍低于正常妊娠组[（26．14±3．12）mmVS（31．05±1．48）mm],但差异无统计学意义（P〉0．05）。结论孕妇外周血中MMP-9、IL-6水平高低及宫颈长度可作为早产发生的预测指标。     

被动吸烟对小鼠白介素-6、白介素-8水平表达的影响

目的观察被动吸烟对小鼠白介素石（IL-6）、白介素-8（IL-8）水平表达的影响。方法将小鼠随机分为2组,分别为实验组和对照组。用ELISA方法检测小鼠血清中IL-6、IL-8的表达情况。结果实验组小鼠血清中IL-6含量为（41．77±1．87）pg／ml,对照组小鼠血清中IL-6含量为（34．98±1．84）pg／ml,两者相比,差异有统计学意义（t=14．15,P〈0．01）。实验组小鼠血清中IL-6含量增高;实验组小鼠血清中IL-8含量为（52．95±1．09）pg／ml,对照组小鼠血清IL-8含量为（65．63±2．59）pg／ml,两者相比差异有统计学意义（t=24．86,P〈0．01）。实验组小鼠血清中IL-8含量减少。结论被动吸烟能促使小鼠IL-6的表达升高,IL-8的表达降低。可能与其参与炎症反应过程有关。     

煤工尘肺患者血清白细胞介素-12和γ-干扰素含量的变化

目的探讨煤工尘肺患者(CWP)血清白细胞介素-12(IL-12)和γ-干扰素(IFN-γ)水平变化及意义。方法用酶联免疫吸附双抗体夹心法检测49名CWP患者(煤工尘肺组)、36名具有相同接尘史但未患CWP的煤矿工人(井下接尘组)及32名非接尘井上健康工人(井上对照组)的血清IL-12和IFN-γ含量。结果煤工尘肺组IL-12水平[(74.0±16.5)pg/ml]明显高于井下接尘组[(25.1±3.6)pg/ml]和井上对照组[(19.8±2.8)pg/ml],组间差异均具有非常显著性(P<0.01);与井上对照组比较,井下接尘组IL-12虽升高,但差异无显著性(P>0.05)。煤工尘肺组、井下接尘组和井上对照组IFN-γ水平无明显改变。Ⅰ期CWP并发肺气肿和Ⅱ期CWP并发肺气肿患者IL-12水平[(58.3±15.6)pg/ml和(62.8±30.9)pg/ml]均高于单纯Ⅰ期和Ⅱ期[(22.9±2.9)pg/ml和(30.2±4.3)pg/ml],两组差异有显著性(P<0.05);Ⅱ期患者血清IL-12水平高于Ⅰ期,但差异无显著性(P>0.05);单纯Ⅱ期和Ⅰ期并发肺气肿患者IFN-γ水平[(13.3±2.3)pg/ml和(21.5±2.8)pg/ml]均低于Ⅰ期[(49.7±7.5)pg/ml],差异具有非常显著性(P<0.01)。煤工尘肺组IL-12和IFN-γ无明显相关性(r=-0.083,P=0.579)。结论IL-12和IFN-γ参与CWP发展过程,与CWP发生和发展均有密切关系。检测IL-12和IFN-γ可能对CWP早期预防及病情监测有一定参考价值。     

生脉散对大肠杆菌脂多糖诱导慢性肝衰竭大鼠细胞因子水平的影响

目的探讨生脉散对慢性肝衰竭大鼠大肠杆菌脂多糖（LPS）诱导细胞因子水平的影响。方法采用CCl。混合液腹腔注射复制慢性肝衰竭大鼠模型,观察LPS诱导2h后生脉散后对血清内毒素、细胞因子水平的影响。结果CCl。混合液能明显增加大鼠血清IL-6[（64．50±18．79）pg／mlVS（4．79±0．57）pg／m1]、ICAM-1[（25100．00±5258．85）pg／mlVS（4215．50±942．79）pg／m1]和TNF-α[（17．55±2．39）pg／mlVS（10．92±5．02）pg／m1]水平（P〈0．05）,但对血清LPS水平无明显影响[（0．058±0．007）EU／mlVS（0．040±0．002）EU／ml,P〉0．05];中药生脉散能显著降低CCl。慢性肝衰竭大鼠血清IL-6、ICAM-1和TNF-α水平[分别为（17．20±3．12）pg／ml、（9490．00±2725．78）pg／ml、（3．00±1．00）pg／ml,P〈0．05]。LPS攻击2h后能明显升高CCI。混合液大鼠血清LPS、TNF-α、IL-6、ICAM-1水平[分别为（0．501±0．019）EU／ml、（19750．00±9655．17）pg／ml、（5615．00±490．50）pg／ml、（41000．00±589．88）pg／ml,P〈0．01]。结论在慢性肝衰竭模型中,LPS能增加TNF-α,IL-6和ICAM-1等炎症因子水平,中药生脉散可降低LPS水平,并抑制LPS诱导的炎症因子水平,阻断了炎性介质及LPS本身对机体的损伤。     

麻风病患者炎症因子水平变化及其临床意义

目的探讨过麻风病的炎症机制。方法对15例麻风病患者血清肿瘤坏死因子-α（TNF-α）、可溶性血管细胞间粘附分子-1（sVCAM-1）和白细胞介素-1（IL-1β）水平进行检测，将其结果与15例本院健康体检者（对照组）比较。结果麻风病组患者血清水平IL-1D、TNF—α和sVCAM-1分别为16．65±4．81（pg／m]）、42．53±18．42（pg／m1）和仔1．53±25．63（ng／m1），对照组分别为8．28±3．99（Pg／m0、26．28±12．32（pg／m]）和46．64±18．58（ng／m]），麻风病组的平均水平较对照组高（P〈0.01）。结论炎症机制参与麻风病患者的发生与发展，巨噬细胞被活化，有多种细胞因子的释放。     

匹多莫德对反复呼吸道感染患儿Th1／Th2细胞因子的影响

目的观察匹多莫德对反复呼吸道感染（RRI）患儿外周血Th1／Th2细胞因子的影响。方法选择RRI患儿60例,随机分为治疗组和对照组,每组各30例。对照组予以常规抗炎、对症治疗。治疗组在对照组治疗基础上,加用匹多莫德口服液（意大利多帕药业有限公司生产,每支7m1）,急性感染期每次7ml,2次／d;急性感染期过后,每次7ml,1次／d;60d为1个疗程。双抗体夹心ELISA法检测两组患儿治疗前后培养上清液单个核细胞（PBMC）中自细胞介素（IL）-4和干扰素γ（IFN-γ）水平的变化。结果治疗组培养上清液PBMC中IL-4水平治疗前为（71．2±10．2）ng／L,治疗后为（19．0±6．2）ng／L（P〈0．01）;IFN-γ水平治疗前为（13．5±1．3）μg／L,治疗后为（69．9±13．0）μg,（P〈0．01）。而对照组治疗前后培养上清液PBMC中IL-4和IFN-γ水平比较差异无统计学意义（P〉0．05）。结论匹多莫德能增强RRI患儿Th1介导的细胞免疫应答,抑制Th2介导的体液免疫应答,调整Th1／Th2细胞因子平衡,临床疗效确切,安全性高,是治疗儿童RRI较理想的药物。     

有氧运动对代谢综合征患者的疗效观察及其机制研究

目的观察长期规律有氧运动对代谢综合征（MS）患者血清瘦素、白介素-18（IL-18）、C反应蛋白（CRP）、可溶性细胞间黏附因子-1（sICAM-1）及胰岛素抵抗指数（HOMA—IR）的影响,并初步探讨有氧运动的作用机制。方法入选40例平时缺乏运动的MS患者,分层随机抽样法分为运动干预组及非诺贝特组,运动干预组进行无氧阈心率水平运动,5次／周,30min/次;非诺贝特组口服非诺贝特胶囊200mg／d。各组干预12周,干预前后采用ELISA法测定血清瘦素、IL-18、CRP、sICAM-1水平。另选20例健康体检者作为正常对照组。结果MS患者血清瘦素[（26．04±9．07）rig／mlVS（8．32±2．94）ng／ml,t=12．72,P〈0．01]、IL-18[（308．27±50．39）pg／mlVS（230．60±29．15）pg／ml,t=6．41,P〈0．01]、CRP[（2．65±0．57）ng／ml vs（1．26±0．23）ng／ml,t=9．69,P〈0．01]、sICAM-1[（331．89±60．08）ng／mlVS（246．43±39．32）ng／ml,t=5．98,P〈0．01]及HOMA—IR（4．38±2．06vs2．12±0．50,t=4．81,P〈0.01）等指标职显高于健康人。经12周运动干预后,MS患者瘦素[（26．38±10．85）ng／mlvs（19．63±6．27）ng／ml,t=2．22,P〈0．05]、IL-18[（309．40±49．77）pg／mlvs（291．80±39．21）pg／ml,t=2．33,P〈0．05]、CRP[（2．73±0．72）ng／mlvs（2．28±0．38）rig／ml,t=3．41,P〈0．01]及sICAM-1[（333．85±55．97）ng／ml V8（306．24±50．55）ng／ml,t=3．16,P〈0．01]水平及HOMA—IR（4．53±2．39V82．89±0．69,t=2．87,P〈0．01）明显下降。结论有氧运动是MS患者一种有效的治疗方法。运动治疗的作用机制可能与降低机体脂肪炎症因子水平,改善血管内皮功能,进一步改善胰岛素抵抗有关。     

Protease-resistant fraction of smoked, dried bonito alleviates atopic dermatitis-like skin lesions in NC/Nga mice

The effect of smoke-dried bonito undigested fraction remaining after microbial protease treatment (SDBR) on a spontaneously occurring mouse model of atopic dermatitis was studied in male 5-wk-old, NC/Nga mice. Smoke-dried bonito, Katsuobushi, is a traditional Japanese food. SDBR contains 2 major components: bonito oil and protease-undigested proteins. Mice were fed a casein diet containing corn oil (C diet) or a diet containing SDBR (SDBR diet) for 18 wk. In comparison with the C diet, the SDBR diet alleviated the increase in skin severity score and plasma IgE concentration in a time-dependent manner, and lowered leucotriene B(4) (LTB(4))-releasing ability upon calcium ionophore A23187 stimulation. The SDBR diet did not affect scratching time. These results demonstrate that SDBR diet alleviates atopic dermatitis-like skin lesions in NC/Nga mice.     

Oral administration of the extract from Hatakeshimeji (Lyophyllum decastes sing.) mushroom inhibits the development of atopic dermatitis-like skin lesions in NC/Nga mice

We examined whether the extract from Hatakeshimeji (Lyophyllum decastes, LD) mushrooms suppresses the development of atopic dermatitis (AD)-like skin lesions induced by repeated application of picryl chloride (PiCl) in NC/Nga mice. Oral administration of LD extract to NC/Nga mice inhibited the development of AD-like skin lesions based on lower total skin severity scores and serum immunoglobulin E (IgE) levels. Splenic lymphocytes were stimulated with the T cell mitogen concanavalin A, and secretion of a Th1 cytokine (IFN-gamma) and a Th2 cytokine (IL-4) was determined by ELISA. IFN-gamma production was not inhibited by treatment with LD extract. On the other hand, IL-4 production was significantly decreased by treatment with LD extract. These results suggest that LD extract exerts anti-allergic actions by suppressing the serum IgE and Th2-type immune responses.     

Anti-Inflammatory and Immune Modulatory Effects of Synbio-Glucan in an Atopic Dermatitis Mouse Model

Many trials have been conducted to treat atopic dermatitis (AD), but these therapies are generally unsuccessful because of their insufficiency or side effects. This study examined the efficacy of β-glucan derived from oats with fermented probiotics (called Synbio-glucan) on an AD-induced mouse model. For the experiment, Nc/Nga mice were exposed to a house dust mite extract (HDM) to induce AD. The mice were placed in one of four groups: positive control group, Synbio-glucan topical treatment group, Synbio-glucan dietary treatment group, and Synbio-glucan topical + dietary treatment group. The experiment revealed no significant difference in the serum IgE concentration among the groups. Serum cytokine antibody arrays showed that genes related to the immune response were enriched. A significant difference in the skin lesion scores was observed between the groups. Compared to the control group tissue, skin lesions were alleviated in the Synbio-glucan topical treatment group and Synbio-glucan dietary treatment group. Interestingly, almost normal structures were observed within the skin lesions in the Synbio-glucan topical + dietary treatment group. Overall, the β-glucan extracted from oats and fermented probiotic mixture is effective in treating atopic dermatitis.     

Di-(2-ethylhexyl) phthalate enhances atopic dermatitis-like skin lesions in mice

Di-(2-ethylhexyl) phthalate (DEHP) has been widely used in polyvinyl chloride products and has become ubiquitous in the developed countries. DEHP reportedly displays an adjuvant effect on immunoglobulin production. However, it has not been elucidated whether DEHP is associated with the aggravation of atopic dermatitis. We investigated the effects of DEHP on atopic dermatitis-like skin lesions induced by mite allergen in NC/Nga mice. NC/Nga male mice were injected intradermally with mite allergen on their right ears. In the presence of allergen, DEHP (0, 0.8, 4, 20, or 100 microg) was administered by intraperitoneal injection. We evaluated clinical scores, ear thickening, histologic findings, and the protein expression of chemokines. Exposure to DEHP at a dose of 0.8-20 microg caused deterioration of atopic dermatitis-like skin lesions related to mite allergen; this was evident from macroscopic and microscopic examinations. Furthermore, these changes were consistent with the protein expression of proinflammatory molecules such as macrophage inflammatory protein-1alpha (MIP-1alpha) and eotaxin in the ear tissue in overall trend. In contrast, 100 microg DEHP did not show the enhancing effects. These results indicate that DEHP enhances atopic dermatitis-like skin lesions at hundred-fold lower levels than the no observed adverse effect level determined on histologic changes in the liver of rodents. DEHP could be at least partly responsible for the recent increase in atopic dermatitis.     

Vitamin E Improves Biochemical Indices Associated with Symptoms of Atopic Dermatitis-Like Inflammation in NC/Nga Mice

We aimed to define whether vitamin E improves biochemical indices associated with symptoms of atopic dermatitis-like inflammation in NC/Nga mice. After picryl chloride (PC) application to their backs, changes in the content of thiobarbituric acid reactive substances (TBARS) and vitamin E, as well as the activity of antioxidant enzymes (superoxide dismutase (SOD), glutathione peroxidase (GSHPx) and catalase) were analyzed in the serum and skin of NC/Nga mice during a symptomatic cycle. The levels of inflammatory factors were also assessed, including IgE, cyclooxigenase-2 (COX-2), tumor necrosis factor (TNF-α) and nuclear factor-κB (NF-κB). When allergic dermatitis was induced by the application of PC to the skin of the mice, skin inflammation appeared 2 wk after PC application, with the peak severity of inflammation observed 5 wk after PC application. Subsequently, the animals recovered from the inflammation by 9 wk after PC application. The TBARS content in the skin and serum increased markedly when the symptoms were the most severe, and decreased to levels near those in control mice by 9 wk after PC application. The activities of SOD and GSHPx in the skin and serum were also positively correlated with symptomatic changes; however, no change in catalase activity was observed 5 wk after PC application. Conversely, vitamin E content decreased at the stage of peak severity. The levels of all inflammatory factors analyzed in this study were altered in a manner similar to other indices. Additionally, vitamin E treatment markedly inhibited these PC-induced alterations. On the basis of these results, it is expected that the observed alterations in biochemical indices, which reflect the symptomatic cycle, may be applicable to objective diagnosis and treatment for atopic dermatitis, and that vitamin E may improve the symptoms of AD.     

2,4-Dinitrofluorobenzene-induced contact hypersensitivity response in NC/Nga mice fed fructo-oligosaccharide

Strategies to manipulate gut microbiota in infancy have been considered to prevent the development of allergic diseases later in life. We previously demonstrated that maternal dietary supplementation with fructo-oligosaccharide (FOS) during pregnancy and lactation modulated the composition of gut microbiota and diminished the severity of spontaneously developing atopic dermatitis-like skin lesions in the offspring of NC/Nga mice. The present study tested whether dietary FOS affects contact hypersensitivity (CHS), another model for allergic skin disease, in NC/Nga mice. In experiment 1, 5-wk-old female NC/Nga mice were fed diets either with or without FOS supplementation for 3 wk and then received 2,4-dinitrofluorobenzene (DNFB) on the ear auricle 5 times at 7-d intervals. FOS supplementation reduced CHS response as demonstrated by ear swelling. Quantitative RT-PCR analysis showed that mRNA levels for interleukin (IL)-10, IL-12p40, and IL-17 in the lesional ear skin were significantly lower in mice fed FOS. In experiment 2, female NC/Nga mice were fed diets either with or without FOS during pregnancy and lactation. After weaning, offspring were fed the diets supplemented with or without FOS. Three weeks after weaning, offspring received DNFB on the ear auricle 4 times at 7-d intervals. Although FOS supplementation after weaning reduced ear swelling, maternal FOS consumption was ineffective in offspring. The present data suggest that dietary FOS reduces CHS while maternal FOS consumption is ineffective in offspring of DNFB-treated NC/Nga mice.     

谷氨酰胺对缺血-再灌注损伤大鼠肠黏膜炎性反应和通透性的影响

目的 研究谷氨酰胺（Gln）对缺血-再灌注损伤大鼠肠黏膜炎性反应和通透性的影响.方法 将48只SD大鼠肠系膜上动脉夹闭造成缺血后恢复血流,建立肠缺血-再灌注损伤模型,将造模后的SD大鼠按随机数字表分为对照组（n=24）和模型＋Gln组（n=24）,两组大鼠肠内营养供给量为热量125.4 kJ/ （kg·d）,氮量0.2g/ （kg·d）,模型＋Gln组喂饲肠内营养加3％ Gln,对照组大鼠喂饲肠内营养加3％大豆蛋白,造模后实验持续8d.检测造模前、造模后、实验第3天和第8天大鼠肠黏膜和血浆核因子-κB （NF-κB）、肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）、Gln、D-乳酸（D-LAC）和二胺氧化酶（DAO）的变化.观察小肠黏膜形态学变化.结果 造模后对照组和模型＋ Gln组大鼠肠黏膜NF-κB表达明显高于造模前（75.0％比0.0％,P=0.013; 70.8％比0.0％,P=0.019）;肠黏膜IL-6明显高于造模前[（313.27±75.28） pg/g比（227.52 ±58.13） pg/g,P=0.023; （321.75±74.46） pg/g比（227.52±58.13） pg/g,P=0.043];肠黏膜TNF-α[（241.28±65.29） pg/g、（240.35 ±64.86） pg/g]明显高于造模前[（172.45±33.76） pg/g,P=0.036,P=0.011];血浆IL-6[（150.32±18.74） ng/L、（148.21 ±20.19） ng/L]明显高于造模前[（116.37±14.59） ng/L,P=0.032,P=0.025];血浆TNF-α[（127.62±14.24） ng/L、（123.86±13.75） ng/L]明显高于造模前[（85.18±8.84） ng/L,P=0.018,P=0.035]; D-LAC[（0.46±0.03） mmol/L、（0.51 ±0.04） mmol/L]明显高于造模前[（0.27±0.02） mmol/L,P=0.041,P=0.018]; DAO[（2.76±0.57） U/ml、（2.58±0.51） U/ml]明显高于造模前[（1.52±0.24） U/ml,P=0.015,P =0.037];而血浆Gln[（0.18±0.01） g/L、（0.21±0.01） g/L]明显低于造模前[（0.39±0.03） g/L,P =0.026,P=0.031].实验第3天和实验第8天,对照组大鼠肠黏膜NF-κB[16例（66.7％）、15例（62.5％）]显著高于造模前[0例（0.0％）,P=0.027,P=0.002];肠黏膜TNF-α[（226.23±55.35） pg/g、（214.76 ±54.82） pg/g]显著高于造模前[（172.45±33.76） pg/g,P=0.042,P=0.038];肠黏膜IL-6[（297.56±71.39） pg/g、（291.49±68.46） pg/g]显著高于造模前[（227.52±58.13） pg/g,P=0.031,P=0.012];血浆IL-6 [（147.38±17.25） ng/L、（144.65±15.32） ng/L]显著高于造模前[（116.37±14.59） ng/L,P=0.016,P=0.034];血浆TNF-α[（121.75±13.72）ng/L、（113.83±11.69） ng/L]显著高于造模前[（85.18±8.84） ng/L,P=0.025,P=0.041];D-LAC[（0.41 ±0.03） mmol/L、（0.53±0.05） mmol/L]显著高于造模前[（0.27±0.02） mmol/L,P=0.029,P=0.030]; DAO [（2.51±0.52） U/ml、（1.76±0.34） U/ml]显著高于造模前[（1.52±0.24） U/ml,P=0.034,P=0.016];但血浆Gln[（0.22±0.01） g/L、（0.21±0.03） g/L]显著低于造模前[（0.39±0.03） g/L,P=0.042,P=0.035].实验第3天模型＋Gln组肠黏膜NF-κB、TNF-α、IL-6 [14例（58.3％）、（213.78±43.76） pg/g、（293.72±69.86） pg/g]明显高于造模前（P=0.038、0.026、0.013）;血浆IL-6、TNF-α、D-LAC、DAO [（135.61 ±14.25） ng/L、（117.35±11.29）ng/L、（0.45 ±0.03） mmol/L、（2.26 ±0.43） U/ml]明显高于造模前（P=0.021、0.032、0.032、0.025）.实验第8天模型＋ Gln组肠黏膜NF-κB、TNF-α、IL-6[9例（37.5％）、（184.53 ±42.16） pg/g、（236.83 ±66.52） pg/g]明显低于造模后和对照组（P=0.024,P=0.027; P=0.026,P=0.039;P =0.013,P=0.028）;血浆IL-6、TNF-α、D-LAC、DAO[（126.35 ±12.74）ng/L、（92.76±9.42）ng/L、（0.31 ±0.02） mmol/L、（1.76 ±0.34） U/ml]明显低于造模后和对照组（P=0.021,P=0.030;P=0.032,P=0.025;P=0.024,P=0.037;P=0.022,P=0.036）,而血浆Gln水平[（0.40±0.03） g/L]明显高于造模后和对照组（P =0.028、0.032）.电镜下可见造模后绒毛、隐窝结构一定程度损害,绒毛稀疏且变短,固有膜内大量炎性细胞浸润,淋巴管扩张、水肿.实验第8天,模型＋Gln组与造模后和对照组比较小肠绒毛、隐窝结构显著恢复;对照组与造模后比较肠黏膜绒毛、隐窝结构恢复不明显,固有膜内仍有炎性细胞浸润.结论 Gln通过调节肠黏膜炎性因子的释放,抑制炎症反应,降低肠黏膜的通透性,修复缺血-再灌注后损伤的肠黏膜.     

Supplemental feeding of a gut microbial metabolite of linoleic acid, 10-hydroxy-cis-12-octadecenoic acid,alleviates spontaneous atopic dermatitis and modulates intestinal microbiota in NC/nga mice

The present study investigated the antiallergic and anti-inflammatory effects of 10-hydroxy-cis-12-octadecenoic acid (HYA), a novel gut microbial metabolite of linoleic acid, in NC/Nga mice, a model of atopic dermatitis (AD). Feeding HYA decreased the plasma immunoglobulin E level and skin infiltration of mast cells with a concomitant decrease in dermatitis score. HYA feeding decreased TNF-α and increased claudin-1, a tight junction protein, levels in the mouse skin. Cytokine expression levels in the skin and intestinal Peyer’s patches cells suggested that HYA improved the Th1/Th2 balance in mice. Immunoglobulin A concentration in the feces of the HYA-fed mice was approximately four times higher than that in the control mice. Finally, denaturing gradient gel electrophoresis of the PCR-amplified 16?S rRNA gene of fecal microbes indicated the modification of microbiota by HYA. Taken together, the alterations in the intestinal microbiota might be, at least in part, associated with the antiallergic effect of HYA.     

Genistein suppresses development of spontaneous atopic-like dermatitis in NC/Nga mice

In this study, we examined the effect of genistein on the severity of dermatitis and level of serum IgE in NC/Nga (NC) mice. NC mice housed in conventional conditions develop spontaneous atopic-like dermatitis; however, oral administration of 20 mg/kg genistein suppresses the development of dermatitis. We also investigated the levels of serum IgE in genistein-treated NC mice and found that the levels were the same as those in control NC mice. We further investigated in vitro IFN-gamma and IL-4 production from spleen cells upon stimulation with anti-CD3 and anti-CD28 mAbs. IFN-gamma production level in NC mice that received 20 mg/kg genistein was significantly lower than that in control NC mice. In contrast, the production level of IL-4 in genistein-treated mice was not significantly different from that in control mice but tended to increase in a dose-dependent manner.