Effects of florfenicol on early cytokine response and survival in a murine septic shock model

氟苯尼考对内毒素休克小鼠生存率和细胞因子的影响@张雪梅$Department of Pharmacology, College of Animal Science and Veterinary Medicine, Jilin University!Changchun 130062, Jilin, China;Department of Animal Medicine, Agricultural College of Yanbian University,     

PACAP inhibit the release and cytokine activity of HMGB1 and improve the survival during lethal endotoxemia

The pathogenesis of sepsis is mediated in part by bacterial endotoxin (lipopolysaccharide; LPS), which stimulates macrophages/monocytes to sequentially release early (e.g., TNF-alpha, IL-1beta) and late [e.g., high mobility group box 1 (HMGB1) protein] pro-inflammatory cytokines. Specifically targeting early mediators has not been effective clinically, in part, because peak mediator activity often has passed before therapy can be initiated. Recent discovery of HMGB1 as a late mediator of lethal sepsis has provided a new target for the treatment of septic shock. Here, we demonstrate that pituitary adenylate cyclase-activating polypeptide (PACAP), an endogenous neuropeptide, significantly attenuated circulating HMGB1 levels and increased survival in animals with established endotoxemia, even if treatment began after acute cytokine response has occurred. In vitro, PACAP suppressed LPS-induced HMGB1 release from macrophages/monocytes, even when given 2-4 h after LPS stimulation. PACAP also suppressed HMGB1 release induced by TNF-alpha or IFN-gamma. Moreover, PACAP inhibits HMGB1-induced cytokine release in vitro and in vivo. These results indicate that PACAP inhibits the release and pro-inflammatory activity of HMGB1 and improves survival during lethal endotoxemia, which confirms this peptide as a candidate for therapy of septic shock.     

Effects of cidofovir treatment on cytokine induction in murine models of cowpox and vaccinia virus infection

Cytokine profiles during cowpox and vaccinia (WR strain) virus infections were characterized in intranasal (i.n.) and intraperitoneal (i.p.) models in BALB/c mice. The time-course of induction and effects of cidofovir treatment on interferon (IFN)-gamma, IFN-gamma inducible protein (IP)-10, interleukin (IL)-6, and monocyte chemoattractant protein (MCP)-1 were determined. The four mouse infection models have distinct patterns of cytokine induction. Cowpox virus i.p. and vaccinia virus i.n. infections showed increased induction throughout the time studied. Cowpox virus i.n. infection resulted in delayed induction of IFN-gamma and IP-10. Cytokine levels were fairly constant during vaccinia virus i.p. infections. Cidofovir treatment (100mg/kg/day i.p. for 2 days) significantly suppressed certain cytokine (IFN- gamma, IL-6, IL-10, IL-11, IP-10, LIF, MCP-1, MCP-3, MCP-5, MIP-1 gamma, and TIMP-1) levels to near normal relative to uninfected animals, as well as prevented mortality and reduced virus titers significantly. Characterization of cytokine responses has implications for understanding the immune responses and pathogeneses of viral infections in these mouse models.     

Effects of montelukast-desloratadine combination on early and late asthma responses

Background: In asthma, montelukast is given on regular basis to reduce eosinophilic airway inflammation. Addition of the antihistaminic desloratadine can amplify the early and late anti-inflammatory activities of montelukast in patients with asthma. Objective: To assess the efficacy data of preventive single-dose desloratadine, montelukast and combination therapy in asthma subjects challenged with inhaled allergens. Methods/results: Analysis of data from a placebo-controlled, cross-over study in which combination preventive therapy was demonstrated to be superior in reducing both early and late allergen-induced inflammatory response when compared with placebo and each component. Conclusion: Although such combination therapy might be effective in preventing the inflammatory flare following the transient allergen exposure, further studies are needed to better define such therapy in terms of duration, doses and therapeutic indications.     

Effects of nano particles on cytokine expression in murine lung in the absence or presence of allergen

Particulate matter (PM) can exacerbate allergic airway diseases. Health effects of PM with a diameter of less than 100 nm, called nano particles, have been focused. We have recently demonstrated that carbon nano particles (14, 56 nm) exaggerate allergic airway inflammation in mice. In the present study, we investigated the effects of repeated pulmonary exposure to carbon nano particles on the expression of a variety of cytokines in the absence or presence of allergen in mice. ICR mice were divided into six experimental groups. Vehicle, two sizes of carbon nano particles, ovalbumin (OVA), and OVA + nano particles were administered intratracheally. Nano particles increased the lung protein levels of thymus and activation-regulated chemokine (TARC), macrophage inflammatory protein (MIP)−1α, and granulocyte-macrophage colony-stimulating factor (GM-CSF) in the absence or presence of allergen. The enhancement was more prominent with 14 nm of nano particles than with 56 nm of nano particles in overall trend. 14 nm nano particle exposure significantly enhanced the lung expressions of interleukin (IL)-2 and IL-10 in the presence of allergen as compared with allergen exposure. These results suggest that pulmonary exposure to nano particles can induce the lung expression of TARC, MIP-1α, GM-CSF in the absence of allergen and can enhance that of TARC, MIP-1α, GM-CSF, IL-2, and IL-10 in the presence of allergen. The enhancing effects are more prominent with smaller particles.     

代谢性内毒素血症小鼠胰腺组织炎性递质的表达

目的观察代谢性内毒素毒血症（ME）小鼠胰腺组织脂联素、白细胞介素-6（IL-6）、单核细胞趋化蛋白-1（MCP-1）及其受体CCR2 mRNA的表达,探讨ME对胰腺组织炎性反应和胰岛素抵抗的影响。方法将纯系C57BL／6J雄性小鼠编号按随机数字法分为代谢性内毒素毒血症组6只和对照组6只,代谢性内毒素毒血症组采用皮下微型泵导入微量脂多糖,建立ME动物模型,对照组则每日泵入相应体积的生理盐水。采用实时荧光定量PCR法的相对定量分析（与GAPDH对照）,检测小鼠胰腺组织脂联素、IL-6、MCP-1及其受体CCR2 mRNA的表达。结果代谢性内毒素毒血症组小鼠空腹血糖浓度降低（P〉0．05）;小鼠体重净增值、空腹胰岛素及胰岛素抵抗指数均升高（均P〈0．05）;小鼠胰腺组织中IL-6、MCP-1及CCR2 mRNA的表达均增强（均P〈0．05）,脂联素mRNA的表达明显减低（P〈0．01）。结论代谢性内毒素血症能够增强胰腺组织中促炎因子的表达,减低抗炎因子的表达,促进胰腺组织的炎症反应,降低胰岛素敏感性。     

三氧化二砷对狼疮小鼠存活率和自身免疫反应的影响(英文)

目的探讨三氧化二砷(ATO)在治疗系统性红斑狼疮中的应用价值并探讨其作用机制。方法① BXSB狼疮小鼠随机分为ATO治疗组和对照组,每组17只。ATO治疗组隔日ip ATO0.4 mg·kg-1至d105,实验持续至d 210结束,观察两组小鼠的存活率,采用ELISA法检测小鼠血清IgG和抗ds-DNA抗体水平。②另外20只BXSB狼疮小鼠,同上分组处理,至d 90处死取脾和肾组织,提取总RNA,用RT-PCR方法检测脾和肾组织中干扰素γ(IFN-γ)mRNA的表达。结果至d210,ATO治疗组小鼠死亡8只,对照组死亡13只。至d90和d105,治疗组小鼠血清抗ds-DNA抗体(A450 nm)分别为0.335±0.011和0.223±0.017,对照组分别为0.688±0.016和0.683±0.014。至d90 ATO治疗组小鼠脾和肾组织IFN-γ mRNA表达较对照组亦明显下降。至d 105,ATO治疗组血清IgG水平较对照组明显下降,分别为(4.9±1.3)和(6.9±1.0)g.L-1。结论 ATO可提高BXSB狼疮小鼠的存活率,降低血清IgG和抗ds-DNA抗体水平,抑制脾和肾组织IFN-γ mRNA的表达。     

Nitrergic modulation of gastrointestinal function during early endotoxemia

After bacterial infection, the host reacts by signalling to the central nervous system where a cascade of physiologic, neuroendocrine and behavioural processes is orchestrated, collectively termed the acute phase response. Endotoxemia following Gram-negative bacterial infection induces a wide array of effects, including fever, loss of appetite and changes in gastrointestinal function that attempt to eliminate the challenge and restore homeostasis. Systemic administration of low doses of endotoxin (5-40 microg/kg) to rats is associated with changes in gastrointestinal motor function, inhibition of gastric acid secretion and increase in the gastric mucosal resistance to damage. These changes are rapid in onset (observed within one hour), not related to vascular dysfunction, and appear to be mediated by mechanisms that involve the peripheral and the central nervous system. Nitric oxide (NO) plays a central role in the physiology of the gastrointestinal tract and its response to illness. Accumulated evidence supports an increase of NO synthesis in the brainstem, as well as in the gastric myenteric plexus thirty minutes after endotoxin administration. Such a synthesis is due to constitutive nitric oxide synthase (NOS) and occurs before the induction of NOS takes place. In this review we provide experimental evidence supporting the hypothesis that activation of a physiologic mechanism, mediated by the autonomic and the central nervous systems as well as constitutive NOS isoforms, is involved in acute changes of gastrointestinal function during early endotoxemia.     

Effects of ketamine on pulmonary inflammatory responses and survival in rats exposed to polymicrobial sepsis.

PURPOSE: Ketamine is reported to suppress production of proinflammatory cytokines and activity of nuclear factor-kappa B (NF-kappaB) after lipopolysaccharide (LPS) stimulation. Our study was designed to investigate the effects of ketamine on pulmonary inflammatory responses and survival in a clinically relevant model of polymicrobial sepsis, induced by cecal ligation and puncture (CLP). METHODS: After the induction of sepsis or sham-operation, animals were treated with ketamine (0.5, 5 or 10 mg/kg) or saline (10 ml/kg) at 3h after operation. At 6 h post-operation, the levels of tumor necrosis factor alpha (TNF-alpha) and interleukin (IL)-6, activity of NF-kappaB, expression of Toll-like receptor 2 (TLR2) and Toll-like receptor 4 (TLR4) of the lungs were measured. And the mortality was recorded for 7 days. RESULTS: TNF-alpha and IL-6 production, NF-kappaB activity, TLR2 and TLR4 expression in rat lungs were increased after CLP. Ketamine at the doses of 5 mg/kg and 10 mg/kg suppressed CLP-induced elevation of TNF-alpha and IL-6 production, NF-kappaB activity and TLR2 expression. Ketamine 0.5, 5 and 10 mg/kg inhibited TLR4 expression in sepsis. Ketamine 5mg/kg and 10 mg/kg after CLP improved the survival of rats. CONCLUSIONS: Ketamine at sub-anesthetic doses could suppress the production of inflammatory cytokines such as TNF-alpha and IL-6, attenuate NF-kappaB activity, and inhibit TLR2 and TLR4 expression in polymicrobial sepsis. These anti-inflammatory effects of ketamine may correlate with improved survival in sepsis.     

Pentamidine blocks the pathophysiologic effects of endotoxemia through inhibition of cytokine release.

Pentamidine isethionate, an antiprotozoal agent with therapeutic value against Pneumocystis carinii pneumonia, has been used for over 30 years without a precise understanding of its mechanism of pharmacologic action. We have previously reported that pentamidine has the capacity to inhibit the release of cytokines from macrophages through a post-translational processing event. The present studies were undertaken to assess the ability of pentamidine to modulate the detrimental effects of murine endotoxemia, a disease with a pathophysiology clearly linked to host-produced cytokines. Under conditions where normal B6C3F1 mice succumbed to the lethal effects of endotoxin, mice pretreated with pentamidine were significantly protected from both mortality and loss of thermoregulatory control. The EC50 for protection from mortality by pentamidine was approximately 11.4 mg/kg. These observations correlated with decreased serum levels of tumor necrosis factor (TNF) and interleukin 6. Inhibition of cytokines was not manifested as part of a generalized inhibition of protein synthesis as demonstrated by the lack of significant modulation of serum albumin in pentamidine-treated animals. In addition to decreased serum concentrations of cytokines, lungs isolated from mice treated with both pentamidine and endotoxin exhibited a decreased release of TNF compared to lungs isolated from mice treated with vehicle and endotoxin. The lower levels of TNF released from lung tissue in pentamidine-treated mice correlated with a lesser degree of alveolar deterioration than was observed in vehicle-treated mice. These data indicate that following endotoxin administration, pentamidine has a protective and antiinflammatory role both systemically and in the lung and suggest that inhibition of inflammatory cytokines may be one mechanism operable in the therapeutic activity of the drug against P carinii pneumonia.     

Physiological antagonism of angiotensin II and lipopolysaccharides in early endotoxemia: pharmacometric analysis

The inhibitory effect of lipopolysaccharides (LPS) on alpha-adrenergic contraction is quite well known, but molecular mechanism of this inhibition is unclear. In the present study, the interaction between alpha-adrenoceptor and vasopressin receptor response, and LPS in rat tail artery was investigated using chemical stimulation. In the presence of LPS, noradrenaline, phenylephrine and arginine-vasopressin, concentration-response curves (CRCs) were shifted to the right with the change in maximal responses. The K(A) and K(B) values calculated in the presence and absence of LPS did not differ significantly. The results strongly suggest that LPS did not change the receptors affinity. The changes in the relationship between receptor occupancy and response to an agonist in the presence of LPS and reduction of K(A)/ED(50) value suggest reduction of receptor reserve. In the presence of angiotensin II (Ang II), CRCs were shifted to the right with significant increase in receptor reserve. Moreover, this effect was still present in LPS-pretreated arteries. The receptor reserve reduced by LPS significantly increased in the presence of Ang II. It suggests that inhibitory effect of LPS is partially reversible. The results strongly suggest that in early endotoxemia, inhibitory effect of LPS may by partially reverted by an increase in activity of renin-angiotensin-aldosterone system.     

Characterization of airway and vascular responses in murine lungs

1. We characterized the responses of murine airways and pulmonary vessels to a variety of endogenous mediators in the isolated perfused and ventilated mouse lung (IPL) and compared them with those in precision-cut lung slices. 2. Airways: The EC50 (microM) for contractions of airways in IPL/slices was methacholine (Mch), 6.1/1.5>serotonin, 0.7/2.0>U46619 (TP-receptor agonist), 0.1/0.06>endothelin-1, 0.1/0.05. In the IPL, maximum increase in airway resistance (RL) was 0.6, 0.4, 0.8 and 11 cmH2O s ml(-1), respectively. Adenosine (< or =1 mM), bombesin (< or =100 microM), histamine (< or =10 mM), LTC4 (< or =1 microM), PAF (0.25 microM) and substance P (< or =100 microM) had only weak effects (<5% of Mch) on RL. 3. Vessels: The EC50 (microM) for vasoconstriction in the IPL was LTC4, 0.06>U46619, 0.05相似文献   

Epidermal cytokines in murine cutaneous irritant responses

Investigations on cytokines in skin irritancy or non-immunological irritant contact dermatitis (ICD) should improve our understanding of their complex mechanism. Numerous studies showed, however, that similar epidermal cytokines have been detected in irritant and allergic reactions, suggesting a lack of specific cytokines that clearly differentiate allergic from irritant reactions even though the pathomechanisms between allergic contact dermatitis (ACD) and ICD are distinguished. Recent data, however, indicate that some mediators may be restricted to allergic responses (contact hypersensitivity). This could provide the impetus to study their implication on irritant reactions. We overview the epidermal cytokines involved in irritant responses compared to those in contact hypersensitivity based on published results of studies using in vitro cell-cultured murine keratinocytes and in vivo murine models.     

微 RNA和去乙酰化在支气管哮喘中对 Th细胞因子的影响

目的探究支气管哮喘( BA)病人外周血 CD4+T淋巴细胞中微 RNA-126(miR-126)、微 RNA-326(miR-326)和微 RNA155(miR-155)以及 Th细胞相关因子的表达情况;探究去乙酰化酶抑制剂曲古抑菌素 A(TSA)对支气管哮喘大鼠 miR-126、miR326和 miR-155以及 Th细胞相关因子的调控作用。方法收集 2015年 10月至 2017年 12月就诊于新疆医科大学第一附属医院、新疆维吾尔自治区人民医院及新疆医科大学附属中医医院的哮喘急性发作期病人( 48例)采用实时荧光定量 PCR技术检测 BA病人 miR-126、miR-326和 miRNA-155以及 Th细胞相关因子 mRNA的表达情况。建立 BA大,鼠模型,分别给予大鼠低、高浓度的 TSA干预, PCR技术检测各组 miR-126、miR-326和 miR-155以及 Th细胞相关因子的表达。免疫组化法检测干扰素 -γ(IFN-γ)、 GATA结合蛋白 3(GATA3)、维甲酸相关孤儿受体( ROR-γ)、叉头翼状螺旋转录因子 3(Foxp3)蛋白表达。结果 BA病人组 miR-126、miR-326和 miR-155 mRNA的表达高于健康人组( 1.63±0.30比 0.54±0.10,2.25±0.67比 1.62±0.39,2.17±0.68比1.75±0.46,均 P<0.05)IFN-γ和 Foxp3 mRNA表达低于健康人组( 0.86±0.19比 1.36±0.27,1.01±0.22比 1.83±0.30,P<0.05)RORγmRNA表达高于正常组(2.79±0.40比 1.62±0.38,P<0.05); BA大鼠模型中,模型组中 miR-126 mRNA的表达高于正常组(2.17±0.39比 0.92±0.06,P<0.01)TSA低剂量组中 miR-126 mRNA的表达低于模型组(2.17±0.39比 1.24±0.20,P<0.05); TSA高剂量组中 GATA3 mRNA的表达低于,正常组和模型组( 3.69±0.88比 1.64±0.35、1.08±0.36,P<0.01);在 TSA高剂量组中 ROR-γ mRNA的表达高于正常组和模型组( 5.50±1.02比 0.89±0.37、10.94±2.85,P<0.05)。结论去乙酰化酶抑制剂可下调 miR-126、miR-326和 miR-155的表达;适量的去乙酰化酶抑制剂可上调细胞因子 IFN-γ和 Foxp3的表达以及下调 GATA3和 ROR-γ的表达。     

双嘧达莫对小鼠免疫功能的影响

双嘧达莫Ｄｉｐ１，１０，４０ｍｇ·ｋｇ－１降低小鼠脾脏系数和末梢血白细胞数，抑制全血白细胞和腹腔巨噬细胞的吞噬功能。降低小鼠血清溶菌酶的含量和血清溶血素的生成，抑制空斑形成细胞的溶血能力。     

Neutral sulfate berberine modulates cytokine secretion and increases survival in endotoxemic mice

AIM: Berberine is thought to be an immunomodulator, so the present study aimed to investigate the effect of berberine on mortality, lung and intestine injury in endotoxemic mice, and the mechanism of its action. METHODS: Mice were challenged with lipopolysaccharide (LPS, 28 mg/kg, ip), and neutral sulfate berberine was administrated intragastrically. Mortality was monitored every 12 h, and histology of the lungs and intestine as well as the plasma tumor necrosis factor-alpha (TNF-alpha), interferon- gamma (IFN-gamma), interleukin-12 (IL-12), IL-10, and nitric oxide (NO) levels were examined. RESULTS: Pretreatment with 50 mg/kg neutral sulfate berberine once a day for 5 days significantly decreased the mortality rate and attenuated tissue injury of the lungs and small intestine in mice challenged with LPS. LPS stimulated a marked increase in plasma levels of TNF-alpha, IFN- gamma, IL-12, IL-10, and NO. The administration of berberine significantly reduced plasma TNF-alpha, IFN- gamma, and NO levels, but did not suppress plasma IL-12 levels in mice exposed to LPS. Furthermore, pretreatment with neutral sulfate berberine augmented IL-10 secretion stimulated by LPS in mice. CONCLUSION: Pretreatment with neutral sulfate berberine attenuates tissue injury and improves survival in endotoxemic mice, which may be mediated, at least in part, by the inhibition of pro-inflammatory mediator production and upregulation of IL-10 release. These findings might provide a new strategy for the treatment of endotoxemia.