Resveratrol Attenuates Heat-Stress-Impaired Immune and Inflammatory Responses of Broilers by Modulating Toll-Like Receptor-4 Signaling Pathway

This study investigated the effect of resveratrol on the immune and inflammatory responses and the mRNA levels of splenic toll-like receptor (TLR)-4 signaling pathway-related genes of broilers under heat stress (HS). One hundred and sixty-two birds were allocated to three groups, each with 6 replicates, for 21 continuous days. The three treatments were as follows: the control group (22 ± 1 °C), the HS (33 ± 1 °C for 10 h d-1 and 22 ± 1 °C for the remaining time) group and the HS + resveratrol (400 mg kg-1) group. At the end of the trial, one bird per replicate close to the average body weight (BW) was selected, exsanguinated, and slaughtered. Compared with the control group, the HS treatment decreased (p<0.05) final BW, average daily gain (ADG), average daily feed intake (ADFI), relative weight of bursa of Fabricius and spleen, serum immunoglobulin (Ig) Y, IgA and interleukin (IL)-10 contents, and splenic IL-10 mRNA level, while it increased (p<0.05) feed/gain, mRNA levels of splenic tumor necrosis factor-α (TNF-α), TLR-4, nuclear factor-kappa-B (NF-κB), IL-1ß, and IL-6. Compared to the HS group, the HS+resveratrol group exhibited increased (p<0.05) final BW, ADG, relative weight of bursa of Fabricius and spleen, serum IgY, IgA and IL-10 contents, and splenic IL-10 mRNA level, while it exhibited lower (p<0.05) TNF-α, IL-1ß and IL-6 contents in serum, and splenic TLR4, TNF-α, IL-1ß, and NF-κB mRNA levels. In conclusion, resveratrol prevented a HS-impairment of the immune function of broilers by blocking the abnormal activation of the TLR4 signaling pathway.(AU)

The effect of olsalazine of chinese generic drugs on ulcerative colitis induced by dextran sulfate sodium salt in BALB/c mice

Purpose: To explore effect and mechanism of olsalazine of Chinese generic drugs on ulcerative colitis induced by dextran sulfate sodium salt (DSS) in BALB/c mice. Methods: The mouse model of ulcerative colitis was induced by free drinking of 3% (w/v) DSS aqueous solution for seven days. The mice were treated with olsalazine (0.6 g·kg-1) of Chinese generic drugs. The therapeutic effect of olsalazine on ulcerative colitis mice was evaluated by measuring disease activity index (DAI), colonic mucosal injury index (CMDI), histopathological score (HS), and detected the expression levels of interleukin (IL)-2, IL-10, tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), IL-1ß in serum and IL-7, IL-17, IL-22, epidermal growth factor (EGF), transforming growth factor ß1 (TGF-ß1) in colonic homogenate of mice. Results: Olsalazine significantly increased the contents of IL-2, IL-10, IL-22, TGF and EGF in ulcerative colitis rats, and significantly decreased the scores of DAI, CMDI, HS and the contents in IL-7, IL-17, TNF-α, IL-1ß and IFN-γ when compared with the model group. It improved the degree of colonic lesion in ulcerative colitis mice. Conclusions: It was suggested that olsalazine has a therapeutic effect on ulcerative colitis induced by DSS in mice, and the mechanism may be related to the increase of IL-2, IL-10, IL-22, TGF, and EGF and the decrease of the expression of IL-7, IL-17, TNF-α, IL-1ß, and IFN-γ.

The Influence of porcine parity on colostrum cytokine levels and their passive transfer to piglets / A influência da paridade suína na diferença dos níveis de citocinas no colostro e na transferência passiva para leitões

The limited ability of newborn piglets to produce cytokines may influence lymphocyte development and response to antigen exposure. As a result, colostrum intake is crucial because it contains nutrients that contribute to immune system development in piglets. Our goal was to investigate the effect of sow parity on the transfer of maternal cytokines to nursing piglets. Sixty piglets from nine sows were divided into six groups: piglets from gilts or sows kept with their dams and allowed to suckle normally; piglets from gilts or sows having their dams exchanged and then allowed to suckle normally; piglets from gilts or sows isolated from their dams and bottle-fed a commercial milk replacer formula for pigs. All piglets remained in the diet groups for 24 hours after birth. Concentrations of cytokines in colostrum and serum of gilt/ sows and serum of piglets were then evaluated. The 13 evaluated cytokines had higher concentrations in colostrum and serum of sows than in gilts. Concentrations of GM-CSF, IFNγ, IL-1α, IL-1ß, IL-2, IL-4, IL-6, IL-10, IL-12, IL-18, and TNFα were higher in piglets suckling sows. Piglets that received commercial formula showed higher concentrations of the cytokines IL1-RA and IL-8 than piglets fed colostrum. This outcome can influence piglets' development into adulthood. In short, our findings demonstrated that maternal parity influenced colostrum cytokine composition and its maternal transfer patterns.(AU)

A capacidade limitada dos leitões recém-nascidos de produzir citocinas pode influenciar o desenvolvimento de linfócitos e a resposta à exposição ao antígeno. Portanto, a ingestão de colostro é importante porque contém nutrientes, que contribuem para o desenvolvimento do sistema imunológico do leitão. O objetivo do estudo foi investigar o efeito da paridade da porca na transferência de citocina materna para leitões lactentes. Sessenta leitões de nove porcas foram divididos em seis grupos: leitões de marrãs/porcas mantidas com suas próprias mães e amamentadas normalmente; leitões de marrãs/porcas que foram trocados de mães e amamentados normalmente; leitões de marrãs/porcas que foram isolados das mães e alimentados com mamadeira com substituto do leite para suínos. Os leitões permaneceram nos grupos por 24 horas após o nascimento. Foram avaliadas as concentrações de citocinas no colostro e plasma das marrãs/porcas e no plasma dos leitões. O colostro e o plasma das porcas apresentaram maiores concentrações das 13 citocinas analisadas do que as marrãs. No mesmo sentido, as concentrações de GM-CSF, IFNγ, IL-1α, IL-1ß, IL-2, IL-4, IL-6, IL-10, IL-12, IL-18 e TNFα foram significantemente maiores nos leitões que mamaram o colostro de porcas. Os leitões que receberam fórmula comercial apresentaram, em especial, concentrações das citocinas IL1-RA e IL-8 superiores aos leitões amamentados com colostro. Isso pode influenciar o desenvolvimento até a fase adulta. Portanto, nossos dados demonstraram que a paridade materna influenciou a composição das citocinas do colostro, bem como as características das citocinas na transferência materna.(AU)

Sheep naturally infected with Anaplasma ovis - evaluation of cardiac and inflammatory biomarkers

Background: Anaplasmosis, also called gall sickness or tropical bovine ehrlichiosis, is an infectious disease caused by species belonging to the genus Anaplasma in domestic and wild animals in tropical and subtropical regions. Anaplasma ovis and A. phagocytophilum are important pathogens of sheep. A. ovis is considered the most common species affecting sheep. The infection is usually subclinical and progresses with high fever, anaemia, icterus, weight loss and abortions. This study aimed to investigate changes in cardiac damage markers, oxidative stress and antioxidant status, cytokines, and acute phase proteins in sheep naturally infected with A. ovis. Materials, Methods & Results: For this purpose, a total of 40 animals, including 20 healthy sheep and 20 sheep infected with anaplasmosis, were used. A. ovis was diagnosed based on clinical findings and peripheral blood smear. Blood smears were prepared from the ear vein. The smears were stained with Giemsa and examined for the presence of Anaplasma spp. Infection was also confirmed by polymerase chain reaction (PCR) analysis. The genomic DNA was isolated from blood, and the MSP-4 gene region was amplified as A. ovis specific target gene. Twenty clinically healthy sheep of the same age group, reared under the same conditions and testing negative in the molecular assessment were used as controls. Blood samples were collected from the cephalic vein and and centrifuged to obtain serum. The serum stored at -20°C until the analysis stage. Serum samples were used for the analysis of cardiac damage markers [troponin I (cTnI), creatine kinase MB (CK-MB), lactate dehydrogenase (LDH) and aspartate transaminase (AST)], oxidative stress parameters [malondialdehyde (MDA), total antioxidant status (TAS), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx)], cytokines [interleukins IL-6, IL-1ß and IL-10, tumour necrosis factor α (TNF-α), and interferon-γ (IFN-γ)] and acute phase proteins [C-reactive protein (CRP), serum amyloid A (SAA) and haptoglobin (Hp)]. cTnI and CK-MB levels were measured using a chemiluminescent immunoassay. MDA, TAS, SOD, CAT, GPx, TNF-α, IL-1ß, IL-6, IL-10, IFN-γ, SAA and Hp levels were measured by an ELISA reader. LDH, AST and CRP levels were measured in an autoanalyzer. cTnI and LDH levels were significantly increased in the infected animals compared to the healthy ones (P < 0.05). The concentration of AST was decreased in infected animals. MDA, TAS, SOD, CAT and GPx levels were significantly increased in the infected animals compared to the healthy ones (P < 0.05). The levels of the inflammatory parameters such as TNF-α, IL-1ß, IL-10 and IFN-γ were significantly increased in the infected animals compared to the healthy ones (P < 0.05). Hp level were significantly increased in the infected group compared with the control group (P < 0.05). However, there was no significant change in CK-MB, SAA and CRP concentrations in the infected animals (P > 0.05). Discussion: Ovine anaplasmosis is an obligate intracellular arthropod disease that causes widespread changes in haematobiochemical, immune response and oxidative stress parameters. Cardiac damage is often overlooked in field conditions due to the lack of adequate knowledge about the pathophysiology of the disease. Our results showed that A. ovis infection leads to significant changes in cardiac biomarkers and that the parasite can cause cardiac dysfunction. This is the first report on cardiac damage markers in Anaplasma-infected sheep. Additionally, the levels of proinflammatory and oxidative stress markers that may cause functional disorders were also found to be increased. Thus, measuring markers of cardiac function, oxidative stress and inflammation can be a useful tool in the early diagnosis of ovine anaplasmosis.

Naringenin attenuates cerebral ischemia/reperfusion injury by inhibiting oxidative stress and inflammatory response via the activation of SIRT1/FOXO1 signaling pathway in vitro

Purpose: To explore the protection of naringenin against oxygen-glucose deprivation/reperfusion (OGD/R)-induced HT22 cell injury, a cell model of cerebral ischemia/reperfusion (I/R) injury in vitro, focusing on SIRT1/FOXO1 signaling pathway. Methods: Cytotoxicity, apoptosis, reactive oxygen species (ROS) generation, malondialdehyde (MDA) content, 4-hydroxynonenoic acid (4-HNE) level, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) activities were measured by commercial kits. Inflammatory cytokines levels were determined by enzyme-linked immunosorbent assay (ELISA). The protein expressions were monitored by Western blot analysis. Results: Naringenin significantly ameliorated OGD/Rinduced cytotoxicity and apoptosis in HT22 cells. Meanwhile, naringenin promoted SIRT1 and FOXO1 protein expressions in OGD/R-subjected HT22 cells. In addition, naringenin attenuated OGD/R-induced cytotoxicity, apoptosis, oxidative stress (the increased ROS, MDA and 4-HNE levels, and the decreased SOD, GSH-Px and CAT activities) and inflammatory response (the increased tumor necrosis factor-α, interleukin [IL]-1ß, and IL-6 levels and the decreased IL-10 level), which were blocked by the inhibition of the SIRT1/FOXO1 signaling pathway induced by SIRT1-siRNA transfection. Conclusion: Naringenin protected HT22 cells against OGD/R injury depending on its antioxidant and anti-inflammatory activities via promoting the SIRT1/FOXO1 signaling pathway.

Effect of Bacillus Subtilis on Immune Function of Hd11 Chicken Macrophages

Poultry is frequently contaminated by Salmonella, a pathogen leading to human health concern worldwide. This study aimed to evaluate the effect of Bacillus subtilis (BS)strain 048 (BS048) on the activation, phagocytosis, sterilization, cytokine secretion, and nitrogen oxide synthesis of HD11 chicken macrophages subjected to Salmonella enteritidis challenge, using lipopolysaccharide treatment as a negative control. The results showed: (1) BS048 had no significant effect on extracellular lactate dehydrogenase activity (p>0.05), while lipopolysaccharide treatment significantly increased extracellular lactate dehydrogenase activity (p >0.05), while lipopolysaccharide treatment significantly increased extracellular lactate dehydrogenase activity (p 0.05);(2)BS048 significantly upregulated the expression levels of pro-inflammatory cytokines (interleukin (IL)- 1â and IL-6), anti-inflammatory cytokines(IL-10 and transforming growth factor-â1), and anti-viral cytokine, interferon-â (p<0.01); ; (3) BS048 significantly upregulated the mRNA expression level of the inducible nitric oxide synthase and its activity as well as extracellular nitrogen oxide level (p <0.01). In conclusion, BS048 could improve antiinflammatory and immune functions of HD11 chicken macrophages, without cytotoxic effects on these cells.(AU)

Monitoring periodontal lesions and their effects during pregnancy: microbiological aspects of the oral cavity and amniotic fluid in pregnant ewes / Monitoramento de lesões periodontais e seus efeitos na gestação: aspectos microbiológicos da cavidade bucal e líquido amniótico de ovelhas gestantes

Periodontitis affects the teeth supporting tissues, leading to tooth loss and damage to animal health. Evidence in humans suggests that oral microorganisms spread systemically, increasing the risk of pregnancy disorders such as miscarriage, prematurity, and low birth weight. This study aimed to verify whether periodontopathogenic microorganisms reach the transplacental unit, culminating in problems in pregnant ewes. After analyzing the oral cavity, 10 clinically healthy pregnant ewes (OGCH group) and 10 pregnant ewes with periodontitis (OGP group) were selected. The subgingival biofilm was collected for the polymerase chain reaction (PCR) test and amniotic fluid for both the PCR and interleukin (IL) analysis. Peripheral blood was collected for complete blood count, and analyses of IL-6, IL1-ß, and tumor necrosis factor-α were performed. Placental fragments were collected to assess the inflammatory changes using optical microscopy. After giving birth, both the ewes and their lambs were weighed. On clinical examination, a positive correlation between bleeding and suppuration (correlation index - CI=0.54), suppuration and marginal gingivitis (CI=0.34), and marginal gingivitis and edema (CI=0.54) was observed. The weights of the ewes (p=0.013) and their respective lambs (p=0.04) in the OGP group were lower than those of their OGCH group counterparts. The hematological analysis revealed that the OGP group ewes showed a slight increase in the mean corpuscular volume (p=0.2447), segmented cells (p=0.3375), and eosinophils (p=0.3823) when compared with the OGCH group ewes, without a statistical difference. Regarding the microorganisms detected in the oral cavity, there was a significant difference between the occurrence of periodontal pockets and the presence of Fusobacterium necrophorum (p=0.0328), Porphyromonas asaccharolytica (p=0.0392), and the Mollicutes class (p=0.0352). Staphylococcus genus (p=0.9107) and Archaea domain (p=0.7245) were detected in the amniotic samples of both groups, without a significant difference, whereas P. asaccharolytica (p=0.2685) was only detected in one sample in the OGCH group. The expression of cytokine IL-6 in the OGP group differed significantly between the prepartum and postpartum periods (p=0.0039); moreover, it differed significantly in the postpartum period between the OGCH and OGP groups (p=0.0198). Histological examination showed a higher percentage of placental changes in the OGP group (70%) than in the OGCH group, such as the presence of macrophages, neutrophils, plasma cells, and multifocal areas of calcification. These results do not corroborate the hypothesis of dissemination of oral microorganisms to the placental unit, suggesting that it constitutes placental isolation in sheep.

A periodontite afeta os tecidos de suporte dos dentes levando à perda dentária e danos à saúde do animal. Evidências em humanos sugerem que os microrganismos orais se espalham sistemicamente, aumentando o risco de distúrbios da gravidez, como aborto espontâneo, prematuridade e baixo peso ao nascer. Este estudo teve como objetivo verificar se microrganismos periodontopatogênicos atingem a unidade transplacentária, culminando em problemas em ovelhas gestantes. Após análise da cavidade oral, foram selecionadas 10 ovelhas gestantes clinicamente saudáveis (grupo OGCH) e 10 ovelhas gestantes com periodontite (grupo OGP). O biofilme subgengival foi coletado para o teste de reação em cadeia da polimerase (PCR) e o líquido amniótico para teste de PCR e análise de interleucina (IL). Sangue periférico foi coletado para hemograma completo e análises de IL-6, IL1-ß e fator de necrose tumoral-α foram realizadas. Fragmentos de placenta foram coletados para avaliação das alterações inflamatórias por meio de microscopia óptica. Após o parto, as ovelhas e seus cordeiros foram pesados. Ao exame clínico, observou-se correlação positiva entre sangramento e supuração (índice de correlação - IC=0,54), supuração e gengivite marginal (IC=0,34) e gengivite marginal e edema (IC=0,54). Os pesos das ovelhas (p=0,013) e de seus respectivos cordeiros (p=0,04) do grupo OGP foram inferiores aos do grupo OGCH. A análise hematológica revelou que as ovelhas do grupo OGP apresentaram discreto aumento no volume corpuscular médio (p=0,2447), células segmentadas (p=0,3375) e eosinófilos (p=0,3823) quando comparadas com as ovelhas do grupo OGCH, sem diferença estatística diferença. Em relação aos microrganismos detectados na cavidade oral, houve diferença significativa entre a ocorrência de bolsas periodontais e a presença de Fusobacterium necrophorum (p=0,0328), Porphyromonas asaccharolytica (p=0,0392) e da classe Mollicutes (p=0,0352). O gênero Staphylococcus (p=0,9107) e o domínio Archaea (p=0,7245) foram detectados nas amostras amnióticas de ambos os grupos, sem diferença significativa, enquanto P. asaccharolytica (p=0,2685) foi detectado apenas em uma amostra do grupo OGCH. A expressão da citocina IL-6 no grupo OGP diferiu significativamente entre os períodos pré e pós-parto (p=0,0039); além disso, diferiu significativamente no período pós-parto entre os grupos OGCH e OGP (p=0,0198). O exame histológico mostrou maior porcentagem de alterações placentárias no grupo OGP (70%) do que no grupo OGCH, como a presença de macrófagos, neutrófilos, plasmócitos e áreas multifocais de calcificação. Esses resultados não corroboram a hipótese de disseminação de microrganismos orais para a unidade placentária, sugerindo que se trata de um isolamento placentário em ovinos.

Cytotoxicity of isolated compounds from Picrasma crenata (Vell) Engl. in animal tumor cell (HTC) / Citotoxicidade de compostos isolados de Picrasma crenata (Vell) Engl. em células tumorais animais (HTC)

The study aim was to evaluate the cytotoxic activity, using the MTT test [3-(4,5-Dimethilthiazol-2-yl)-2,5-diphenil tetrazolium bromide], from the crude extract of Picrasma crenata (Pau Tenente) and its isolated compounds, quassin and parain, in culture of rat liver tumor cells (HTC). The test was carried out exposing the cells for 24, 48 and 72 hours to concentrations of 5, 10, 50, 100, 200, 300, 400, 500 and 1000 µg of crude extract of Pau Tenente/mL of culture medium and 1, 5, 10, 15, 20, 40, 60, 80 and 100 µg of quassin or parain compounds/mL of culture medium. The absorbances averages results obtained showed that the crude extract did not present cytotoxicity for the HTC cells in all the concentrations and evaluated times. For quassin, the concentrations of 80 and 100 µg/mL were cytotoxic, after 72 hours of treatment. For parain, the concentrations of 1, 5, 20, 40, 60, 80 and 100 µg/mL, in 72 hours, were cytotoxic, revealing a new activity for this compound. Thus, the results demonstrate a first indication of the cytotoxic activity of compounds quassin and parain, adding an important social and economic value to them, and may have application in future research and in pharmaceutical industry.

O objetivo do estudo foi avaliar a atividade citotóxica, por meio do teste MTT [brometo de 3-(4,5-dimetiltiazol-2-il)-2,5-difenil tetrazólio], do extrato bruto de Picrasma crenata (Pau Tenente) e seus compostos isolados, quassina e paraína, em cultura de células tumorais de fígado de rato (HTC). O teste foi realizado expondo as células por 24, 48 e 72 horas às concentrações de 5, 10, 50, 100, 200, 300, 400, 500 e 1000 µg de extrato bruto de Pau Tenente/mL de meio de cultura e 1, 5, 10, 15, 20, 40, 60, 80 e 100 µg de compostos de quassina ou paraína/mL de meio de cultura. Os resultados das médias de absorbâncias obtidos mostraram que o extrato bruto não apresentou citotoxicidade para as células HTC em todas as concentrações e tempos avaliados. Para quassina, as concentrações de 80 e 100 µg/mL foram citotóxicas, após 72 horas de tratamento. Para a paraína, as concentrações de 1, 5, 20, 40, 60, 80 e 100 µg/mL, em 72 horas, foram citotóxicas, revelando uma nova atividade para este composto. Assim, os resultados demonstram uma primeira indicação da atividade citotóxica dos compostos quassina e paraína, agregando-lhes importante valor social e econômico, podendo ter aplicação em pesquisas futuras e na indústria farmacêutica.

Propolis anti-inflammatory effects on MAGE-1 and retinoic acid-treated dendritic cells and on Th1 and T regulatory cells

Background: Propolis exhibits huge potential in the pharmaceutical industry. In the present study, its effects were investigated on dendritic cells (DCs) stimulated with a tumor antigen (MAGE-1) and retinoic acid (RA) and on T lymphocytes to observe a possible differential activation of T lymphocytes, driving preferentially to Th1 or Treg cells. Methods: Cell viability, lymphocyte proliferation, gene expression (T-bet and FoxP3), and cytokine production by DCs (TNF-α, IL-10, IL-6 and IL-1ß) and lymphocytes (IFN-γ and TGF-ß) were analyzed. Results: MAGE-1 and RA alone or in combination with propolis inhibited TNF-α production and induced a higher lymphoproliferation compared to control, while MAGE1 + propolis induced IL-6 production. Propolis in combination with RA induced FoxP3 expression. MAGE-1 induced IFN-γ production while propolis inhibited it, returning to basal levels. RA inhibited TGF-ß production, what was counteracted by propolis. Conclusion: Propolis affected immunological parameters inhibiting pro-inflammatory cytokines and favoring the regulatory profile, opening perspectives for the control of inflammatory conditions.(AU)

Effect of Turmeric (Curcuma Longa) on Duodenal Structure in Broiler Chickens

Supplementation of feed with turmeric (Curcuma longa) has been shown to be beneficial in poultry farming. The present study aimed at evaluating the effect of turmeric on the duodenal structure of broiler chickens during the first 42 days of life. A control and three treatment groups (turmeric powder added to feed at the following doses: E1 - 5; E2 - 10; E3 - 20 g/kg feed) were constituted (n = 8). Addition of turmeric powder in the feed resulted in an increase in intestinal villi height and a decrease in crypts depth in case of groups E1 and E2, while the villus height to crypt depth ratio generally did not differ significantly from the control. Turmeric also influenced the histological structure of the duodenum, as well as the presence of IL-6 and TNFα, as evidenced through staining. Addition of 0.5 and 1 % turmeric powder in the feed had evident results on body weight gain.(AU)

Cyclosporine A increases the intensity of Toxocara canis infection in swiss mice / Ciclosporina A aumenta a intensidade da infecção por Toxocara canis em camundongos suíços

Toxocariasis is a zoonotic disease of worldwide distribution. The connection between parasitic diseases and conditions that depress the immune system, such as the use of immunosuppressive drugs, has been studied. The purpose of this study was to evaluate the effect of Cyclosporine A (CsA) on the intensity of infection, humoral response and gene transcription of interleukins IL-4, IL-10 and IL-12 in mice experimentally infected with Toxocara canis. To this end, mice were divided into two groups treated with CsA (G1: 10 mg/Kg and G2: 50 mg/kg), the G3 and G4 group received PBS. After the last administration of the drug or PBS (orally every 48 hours for 15 days), groups G1, G2 and G3 were inoculated with 1200 eggs of T. canis. Was collected blood samples on days zero, 15 and 30 days post-inoculation (PI), for ELISA test and the mice were euthanized 30 days PI. The organs and striated muscle tissue were collected for the recovery of larvae. The splenocytes were analyzed by RT-PCR. The intensity of infection in the mice treated with 50 mg/kg of CsA was 65.5% higher than in the control group (p=0.001). An analysis of the kinetics of anti-Toxocara antibody revealed that the groups treated with CsA showed significantly higher mean levels of antibodies on day 15 PI. The transcription of the three tested interleukins showed no statistical difference between G2 and G3 (control). It was concluded that the immunosuppression triggered by CsA (50 mg/Kg) favored the establishment of a larger number of T. canis larvae without, however, altering immunoglobulin production and IL-4, IL-10 and IL-12 transcription on day 30 PI.

A toxocaríase é uma zoonose de distribuição mundial. A conexão entre doenças parasitárias e condições que deprimem o sistema imunológico, como o uso de drogas imunossupressoras, tem sido estudada. O objetivo deste estudo foi avaliar o efeito da Ciclosporina A (CsA) na intensidade da infecção, resposta humoral e transcrição gênica das interleucinas IL-4, IL-10 e IL-12 em camundongos experimentalmente infectados com Toxocara canis. Para tanto, os camundongos foram divididos em dois grupos tratados com CsA (G1: 10 mg/Kg e G2: 50 mg/kg), os grupos G3 e G4 receberam PBS. Após a última administração da droga ou PBS (via oral a cada 48 horas por 15 dias), os grupos G1, G2 e G3 foram inoculados com 1200 ovos de T. canis. Foram coletadas amostras de sangue nos dias zero, 15 e 30 dias pós-inoculação (PI), para teste de ELISA e os camundongos foram eutanasiados 30 dias PI. Os órgãos e tecido muscular estriado foram coletados para a recuperação das larvas. Os esplenócitos foram analisados ​​por RT-PCR. A intensidade da infecção nos camundongos tratados com 50 mg/kg de CsA foi 65,5% maior do que no grupo controle (p=0,001). Uma análise da cinética do anticorpo anti-Toxocara revelou que os grupos tratados com CsA apresentaram níveis médios de anticorpos significativamente maiores no dia 15 PI. A transcrição das três interleucinas testadas não apresentou diferença estatística entre G2 e G3 (controle). Concluiu-se que a imunossupressão desencadeada pela CsA (50 mg/Kg) favoreceu o estabelecimento de um maior número de larvas de T. canis sem, no entanto, alterar a produção de imunoglobulinas e a transcrição de IL-4, IL-10 e IL-12 no dia 30 PI.

Acute and chronic immunomodulatory response mechanisms against Toxocara canis larvae infection in mice / Mecanismos da resposta imunomoduladora aguda e crônica contra a infecção por larvas de Toxocara canis em camundongos

The objective of this work was to evaluate the early and late immunological modulation of an experimental infection of T. canis larvae in mice. Mice were infected with 100 infective larvae and euthanized at different period: 24, 48 hours post infection (HPI), 15- and 30 days post infection (DPI). The humoral response was evaluated by indirect ELISA. Quantitative RT-PCR (qPCR) was used to quantify the mRNA transcription of cytokines IL4, IL10, IL12 and Ym1 in the early and late infection periods. Infection with T. canis was able to generate specific total IgG at 15- and 30- DPI. Analyzing the IgG isotype revealed a significant differentiation for IgG1 compared with IgG2a, IgG2b and IgG3, characterizing a Th-2 response. Evaluating the gene transcription at the early phase of infection, higher transcription levels of IL10, IL4 and Ym1 and a downregulation of IL12 were observed. By the late phase, increased transcription levels of IL4, Ym1 and IL12 were observed, and downregulation of IL-10 transcription was observed. The data obtained suggest that during experimental infection with T. canis, the participation of the IL4, IL10, IL12 cytokines and Ym1 can play an important role in T. canis immunomodulation.(AU)

O objetivo deste trabalho foi avaliar a modulação imunológica precoce e tardia da infecção experimental de larvas de T. canis em camundongos. Estes foram infectados com 100 larvas infectantes e eutanasiados em diferentes períodos: 24 e 48 horas pós infecção (HPI), 15 e 30 dias após a infecção (DPI). A resposta humoral foi avaliada por ELISA indireto. RT-PCR quantitativo (qPCR) foi usado para quantificar a transcrição de mRNA das citocinas IL4, IL10, IL12 e Ym1 nos períodos de infecção precoce e tardia. A infecção por T. canis foi capaz de gerar IgG total específico aos 15 e 30 DPI. A análise do isótipo IgG revelou uma diferenciação significativa para IgG1 em comparação com IgG2a, IgG2b e IgG3, caracterizando uma resposta Th-2. Avaliando-se a transcrição gênica na fase inicial da infecção, foram observados níveis mais elevados de transcrição de IL10, IL4 e Ym1 e a regulação negativa de IL12. Na fase tardia, foram observados níveis aumentados de transcrição de IL4, Ym1 e IL12, e foi observada regulação negativa da transcrição de IL-10. Os dados obtidos sugerem, que durante a infecção experimental com T. canis, a participação das citocinas IL4, IL10, IL12 e Ym1 podem desempenhar um papel importante na imunomodulação de T. canis.(AU)

Effects of N-acetylcysteine on the inflammatory response and bacterial translocation in a model of intestinal obstruction and ischemia in rats

Purpose: To evaluate effect of N-acetylcysteine (NAC) associated with Ringer lactate or hypertonic saline in inflammation and bacterial translocation on experimental intestinal obstruction (IO). Methods: Wistar rats was subjected to IO. Six or 24 hours after, rats were subjected to enterectomy and fluid resuscitation: IO, RL (subjected to the same procedures but with fluid resuscitation using Ringer's lactate solution); RLNAC (added NAC to Ringer's solution); and HSNAC (surgical procedure + fluid reposition with 7.5% hypertonic saline and NAC). After 24 h, tissues were collected to cytokines, bacterial translocation, and histological assessments. Results: In kidney, interleukin-1beta (IL-1beta) was lower in the groups with fluid resuscitation compared to IO group. The RLNAC showed lower levels compared to the RL. Interleukin-6 (IL-6), interleukin-10 (IL-10), tumor necrosis factor-alpha (TNF-alpha), and (IFN-gamma) were lower in the treatment groups than in IO. In lung, IL-1beta and IL-6 were lower in RLNAC compared to IO. IL-10 was lower in RL, RLNAC and HSNAC compared to IO. TNF-alpha was higher in HSNAC compared to both RL and RLNAC. Bacterial translocation was observed in all animals of IO group. In kidneys, inflammation and congestion degrees were lower in HSNAC compared to RL. In lungs, inflammation levels were higher in RLNAC compared with the sham group. Conclusions: The data indicates that NAC associated with RL can promote a decrease in the inflammatory process in the kidneys and lungs in rats, following intestinal obstruction and ischemia in rats.

Photosynthetic metabolism and antioxidant in Ormosia arborea are modulated by abscisic acid under water deficit? / Metabolismo fotossintético e antioxidante em mudas de Ormosia arborea são modulados pelo ácido abscísico sob deficit hídrico?

The aim of this research was to evaluate the effect of abscisic acid (ABA) on gas exchange and the activity of antioxidant enzymes of Ormosia arborea (Vell.) Harms seedlings under water deficit and its influence on the recovery potential of the seedlings. The experiment was conducted using four treatments, being daily irrigation or water restriction without and with 10 µM ABA. Seedlings under water deficit + ABA showed greater adjustment to drought, and when re-irrigated, they restored photosynthetic metabolism and water potential. ABA minimizes the reduction in the photosynthetic metabolism and water potential of the leaf, however, it does not increase the antioxidant activity of the O. arborea seedlings under water deficit. These results suggest that this species exhibits plasticity, which enables it to survive also in environments subjected to temporary water deficit regardless of the supplementation of ABA. We suggest that other doses of ABA be researched to expand the beneficial effect of ABA on this species.

O objetivo deste trabalho foi avaliar o efeito do ácido abscísico (ABA) nas trocas gasosas e na atividade de enzimas antioxidantes de mudas de Ormosia arborea (Vell.) Harms sob deficiência hídrica e sua influência no potencial de recuperação das mudas. O experimento foi conduzido com quatro tratamentos, sendo eles irrigação diária ou restrição hídrica sem e com 10 µM ABA. As mudas sob déficit hídrico + ABA apresentaram maior ajuste à seca e ao serem re-irrigadas restabeleceram o metabolismo fotossintético e o potencial hídrico. O ABA minimizou a redução do metabolismo fotossintético e do potencial da água na folha, porém, não aumentou a atividade antioxidante de mudas de O. arborea sob déficit hídrico. Esses resultados sugerem que esta espécie apresenta plasticidade fisiológica, o que lhe permite sobreviver em ambientes sujeitos a déficit hídrico temporário, independente da suplementação de ABA. Sugerimos que outras doses de ABA sejam avaliadas para ampliar os efeitos benéficos do ABA sobre esta espécie.

Abscisic acid and in vitro conservation of Manihot wild species

Abscisic acid (ABA) is associated with bud dormancy, leaf abscission, and germplasm growth inhibition in in vitro conservation. We evaluated the effects of ABA in four wild Manihot accessions and one cassava accession (M. esculenta Crantz) to refine in vitro conservation methods for these species. The experiment was performed at the Laboratory for Tissue Culture from Embrapa, Cruz das Almas, Bahia State, Brazil. The statistical design was completely random in a 5 × 5 factorial scheme [(5 ABA dosages (0, 0.25, 0.50, 0.75, and 1 mg L-1) and 5 Manihot species (M. pseudoglaziovii, M. tristis, M. flabellifolia, M. chlorosticta, and M. esculenta)], with 15 replicates. Mini-cuttings of 1 cm were used, each inoculated in 10 mL of modified Murashige and Skoog medium, solidified with Phytagel® (2.4 g L-1) containing the respective ABA dosages. Tubes containing these mini-cuttings were placed in a germplasm conservation room with an irradiance of 30 µmol m-2 s-1, temperature of 22 ± 1°C, and photoperiod of 12 hours. Plant height (cm), the number of living and senescent leaves, shoots, and mini-cuttings (1 cm), and fresh and dry weights of shoots and roots (mg) were evaluated after 150 days. Growth reduction was prominent in M. pseudoglaziovii, M. tristis, and M. flabellifolia during the in vitro conservation period. In the present study, the addition of ABA did not promote the expected reduction in plant growth.(AU)

Application of abscisic acid (S-ABA) at different stages of ripening on color development of 'Rubi' table grape / Aplicação do ácido abscísico (S-ABA) durante a maturação no desenvolvimento da cor da uva fina de mesa 'Rubi'

The color of the berries is an important aspect of the quality of table grapes and crucial for marketing. The 'Rubi' table grapes grown in the subtropical climate generally lack color intensity due to the inhibition of anthocyanins by high temperatures during ripening. The exogenous application of abscisic acid (S-ABA) can be used to overcome this problem as the accumulation of anthocyanins in the berry skin is regulated by this plant growth regulator. The objective of this study was to assess the effect of the exogenous application of S-ABA at different stages of ripening on color development in 'Rubi' table grapes using the soluble solids (SS) content as a marker of ripening. The study was conducted during two seasons in commercial vineyards. The first trial was conducted in Marialva, Parana, Brazil, during the 2019 summer season crop (harvest in December). S-ABA (400 mg L-1) was exogenously applied at different stages of ripening of 'Rubi' table grapes (determined by the SS content of the berries): control (without application); SS = 8-9 ºBrix; SS = 10-11 ºBrix; SS = 8-9 ºBrix (two applications; the second one applied 10 days after the first); and SS = 10-11 ºBrix (two applications; the second one applied 10 days after the first). The second trial was conducted in Cambira, Parana, Brazil, during the 2020 off-season crop (harvest in May). S-ABA (400 mg L-1) was exogenously applied at different stages of ripening of 'Rubi' grapes: control (without application); SS = 6-7 ºBrix; SS = 7-8 ºBrix; SS = 9-10 ºBrix; SS = 6-7 ºBrix (two applications; the second one applied 14 days after the first); SS = 7-8 ºBrix (two applications; the second one applied 14 days after the first); and SS = 9-10 ºBrix (two applications; the second one applied 14 days after the first).(AU)

A cor das bagas é um aspecto importante de qualidade e comercialização de uvas de mesa. A uva fina de mesa 'Rubi' produzida em clima subtropical geralmente apresenta deficiência de cor devido à inibição da formação de antocianinas pelas altas temperaturas durante a maturação. Um dos métodos utilizados para superar esse problema é a aplicação exógena de ácido abscísico (S-ABA), uma vez que o acúmulo de antocianinas na casca das bagas é controlado por esse regulador vegetal. O objetivo deste trabalho foi determinar o efeito da aplicação do S-ABA em diferentes fases de maturação no desenvolvimento da cor da uva fina de mesa 'Rubi', visando o incremento e uniformização da cor dos seus cachos, tendo como referência o teor de sólidos solúveis (SS) das bagas. O experimento foi avaliado em dois ensaios a campo, em pomares comerciais. O primeiro ensaio foi realizado em Marialva, Paraná, Brasil, durante a safra de verão (colheita em dezembro) de 2019. Os tratamentos foram constituídos da aplicação de S-ABA na concentração de 400 mg L-1, em diferentes fases de maturação da uva 'Rubi', de acordo com o teor de SS das bagas, assim descritos: controle (sem aplicação); SS = 8-9 ºBrix; SS = 10-11 ºBrix; SS = 8-9 ºBrix em duas aplicações, sendo a segunda realizada aos 10 dias após a primeira; e SS = 10-11 ºBrix, em duas aplicações, sendo a segunda realizada aos 10 dias após a primeira. O segundo ensaio foi realizado em Cambira, Paraná, Brasil, durante a safra temporona ou de outono (colheita em maio) de 2020. Os tratamentos foram constituídos da aplicação de S-ABA na concentração de 400 mg.L-1 em diferentes fases de maturação da uva 'Rubi', de acordo com o teor de SS das bagas, assim descritos: controle (sem aplicação); SS = 6-7 ºBrix; SS = 7-8 ºBrix; SS = 9-10 ºBrix; SS = 6-7 ºBrix em duas aplicações, sendo a segunda realizada aos 14 dias após a primeira; SS = 7-8 ºBrix em duas aplicações, sendo a segunda realizada aos 14 dias após a primeira; e SS = 9-10 ºBrix em duas aplicações, sendo a segunda realizada aos 14 dias após a primeira. O delineamento experimental de ambos experimentos foi em blocos casualizados com 4 repetições, sendo que cada parcela foi composta por uma videira. As variáveis analisadas foram antocianinas totais, índice de cor (CIRG), e atributos (L*, C*, h° e ΔE) das bagas.(AU)

Phytochemical characterization and biological activities of Plectranthus barbatus Andrews

Abstract Plectranthus barbatus Andrews (Lamiaceae) is widely distributed in the world and has a range of popular therapeutic indications. This work aimed to evaluate the phytochemical characterization of two leaf extracts of P. barbatus, and their antimicrobial, antineoplastic and immunomodulatory potential. After collection, herborization and obtainment of the P. barbatus aqueous extract (PBA) and acetone:water 7:3 P. barbatus organic extract (PBO), the phytochemical characterization was performed by high-performance liquid chromatography (HPLC). The antimicrobial activity was performed to determine the minimum inhibitory concentration (MIC) against eight bacterial strains using the microdilution test and the fungus Trichophyton rubrum by disc diffusion assay and microdilution test. Cytotoxicity was assessed by MTT and trypan blue methods in normal peripheral blood mononuclear cells (PBMCs) at concentrations ranged between 0.1 to 100 µg.mL-1 and in neoplastic cell lines Toledo, K562, DU-145 and PANC-1 at 1, 10 and 100 µg.mL-1 . Immunomodulatory activity, was evaluated by sandwich ELISA of proinflammatory cytokines at BALB/c mice splenocytes cultures supernatant. Both extracts presented flavonoids, cinnamic derivatives, steroids and ellagic acid. PBO showed bacteriostatic activity against Acinetobacter baumannii (MIC = 250 µg.mL-1) clinical isolate and PBA fungistatic activity against Trichophyton rubrum (MIC = 800 µg.mL-1). The extracts did not exhibit toxicity to PBMCs and neoplastic cells (IC50 > 100 µg.mL-1). Additionally, PBO at 100 µg.mL-1 significantly inhibited IFN- and IL-17A cytokines (p = 0.03). Plectranthus barbatus is a potential candidate for therapeutic use due to its low toxicity in healthy human cells and exhibits biological activities of medical interest as bacteriostatic, fungistatic and immunomodulatory.

Resumo Plectranthus barbatus Andrews (Lamiaceae) é amplamente distribuída no mundo e com uma série de indicações terapêuticas populares. Este trabalho teve como objetivo avaliar a caracterização fitoquímica de dois extratos da folha de P. barbatus e seu potencial antimicrobiano, antineoplásico e imunomodulador. Após coleta, herborização e obtenção do extrato aquoso (PBA) e acetona: água 7: 3 (orgânico) (PBO) de P. barbatus, a caracterização fitoquímica foi realizada por cromatografia líquida de alta eficiência (CLAE). A atividade antimicrobiana foi realizada para determinar a concentração inibitória mínima (CIM) contra oito cepas bacterianas usando o teste de microdiluição e o fungo Trichophyton rubrum por ensaio de difusão em disco e teste de microdiluição. A citotoxicidade foi avaliada por métodos MTT e azul de tripan em células normais mononucleares do sangue periférico (CMSP) em concentrações variadas entre 0,1 a 100 µg.mL-1 e nas linhagens celulares neoplásicas Toledo, K562, DU-145 e PANC-1 em 1, 10 e 100 µg.mL-1 . A atividade imunomoduladora foi avaliada por ELISA sanduíche de citocinas pró-inflamatórias em sobrenadante de culturas de esplenócitos de camundongos BALB/c. Ambos os extratos apresentaram flavonoides, derivados cinâmicos, esteróides e ácido elágico. O PBO mostrou atividade bacteriostática contra Acinetobacter baumannii (CIM = 250 µg.mL-1) e atividade fungistática do PBA contra Trichophyton rubrum (CIM = 800 µg.mL-1). Os extratos não apresentaram toxicidade para CMSP e células neoplásicas (IC50 > 100 µg.mL-1). Além disso, o PBO a 100 µg.mL-1 inibiu significativamente as citocinas IFN- e IL-17A (p = 0,03). Plectranthus barbatus é um candidato potencial para uso terapêutico devido à sua baixa toxicidade em células humanas saudáveis e exibe atividade de interesse médico como bacteriostática, fungistática e imunomoduladora.

Kukoamine A activates Akt/GSK-3ß signaling pathway to inhibit oxidative stress and relieve myocardial ischemia-reperfusion injury

Purpose: Myocardial ischemia/reperfusion (MI/R) injury refers to a pathological condition of treatment of myocardial infarction. Oxidative stress and inflammation are believed to be important mechanisms mediating MI/R injury. Kukoamine A (KuA), a sperm, is the main bioactive component extracted from the bark of goji berries. In this study, we wanted to investigate the possible effects of KuA on MI/R injury. Methods: In this experiment, all rats were divided into sham operation group, MI/R group, KuA 10 mg + MI/R group, KuA 20 mg + MI/R group. After 120 min of ischemia/reperfusion treatment, left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP), maximal rates of rising and fall of left ventricular pressure (±dp/dtmax), and ischemic area were detected. Serum samples of rats in each group were collected. The enzyme activities of catalase (CAT), glutathione peroxidase (GSH-PX), superoxide dismutase (SOD), levels of malondialdehyde (MDA), CK muscle/brain (CK-MB), tumor necrosis factor (TNF), interleukin-1ß (IL-1ß), and interleukin-6 (IL-6) were detected using enzyme-linked immunosorbent assay (ELISA). The apoptosis of myocardium in each group was detected according to the instructions of the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. The expressions of mammalian target of glycogen synthase kinase-3ß (GSH-3ß) and protein kinase B (Akt) mRNA level in myocardial tissues were detected via reverse transcription-polymerase chain reaction (RT-PCR). Results: MI/R rats showed a significant increase in oxidative stress and inflammation. In addition, we showed that KuA significantly improved the myocardial function such as LVSP, left ventricular ejection fraction, +dp/dt, and -dp/dt. Here, it attenuated dose-dependent histological damage in ischemia-reperfused myocardium, which is associated with the enzyme activities of SOD, GSH-PX, and levels of MDA, IL-6, TNF-α, L-1ß. Conclusions: KuA inhibited gene expression of Akt/GSK-3ß, inflammation, oxidative stress and improved MR/I injury. Taken together, our results allowed us to better understand the pharmacological activity of KuA against MR/I injury.

Phytochemical characterization and biological activities of Plectranthus barbatus Andrews / Caracterização fitoquímica e atividades biológicas de Plectranthus barbatus Andrews

Plectranthus barbatus Andrews (Lamiaceae) is widely distributed in the world and has a range of popular therapeutic indications. This work aimed to evaluate the phytochemical characterization of two leaf extracts of P. barbatus, and their antimicrobial, antineoplastic and immunomodulatory potential. After collection, herborization and obtainment of the P. barbatus aqueous extract (PBA) and acetone:water 7:3 P. barbatus organic extract (PBO), the phytochemical characterization was performed by high-performance liquid chromatography (HPLC). The antimicrobial activity was performed to determine the minimum inhibitory concentration (MIC) against eight bacterial strains using the microdilution test and the fungus Trichophyton rubrum by disc diffusion assay and microdilution test. Cytotoxicity was assessed by MTT and trypan blue methods in normal peripheral blood mononuclear cells (PBMCs) at concentrations ranged between 0.1 to 100 µg.mL-¹ and in neoplastic cell lines Toledo, K562, DU-145 and PANC-1 at 1, 10 and 100 µg.mL-¹ . Immunomodulatory activity, was evaluated by sandwich ELISA of proinflammatory cytokines at BALB/c mice splenocytes cultures supernatant. Both extracts presented flavonoids, cinnamic derivatives, steroids and ellagic acid. PBO showed bacteriostatic activity against Acinetobacter baumannii (MIC = 250 µg.mL-¹) clinical isolate and PBA fungistatic activity against Trichophyton rubrum (MIC = 800 µg.mL-¹). The extracts did not exhibit toxicity to PBMCs and neoplastic cells (IC50 > 100 µg.mL-¹). Additionally, PBO at 100 µg.mL-1 significantly inhibited IFN-γ and IL-17A cytokines (p = 0.03). Plectranthus barbatus is a potential candidate for therapeutic use due to its low toxicity in healthy human cells and exhibits biological activities of medical interest as bacteriostatic, fungistatic and immunomodulatory.

Plectranthus barbatus Andrews (Lamiaceae) é amplamente distribuída no mundo e com uma série de indicações terapêuticas populares. Este trabalho teve como objetivo avaliar a caracterização fitoquímica de dois extratos da folha de P. barbatus e seu potencial antimicrobiano, antineoplásico e imunomodulador. Após coleta, herborização e obtenção do extrato aquoso (PBA) e acetona: água 7: 3 (orgânico) (PBO) de P. barbatus, a caracterização fitoquímica foi realizada por cromatografia líquida de alta eficiência (CLAE). A atividade antimicrobiana foi realizada para determinar a concentração inibitória mínima (CIM) contra oito cepas bacterianas usando o teste de microdiluição e o fungo Trichophyton rubrum por ensaio de difusão em disco e teste de microdiluição. A citotoxicidade foi avaliada por métodos MTT e azul de tripan em células normais mononucleares do sangue periférico (CMSP) em concentrações variadas entre 0,1 a 100 µg.mL-¹ e nas linhagens celulares neoplásicas Toledo, K562, DU-145 e PANC-¹ em 1, 10 e 100 µg.mL-¹ . A atividade imunomoduladora foi avaliada por ELISA sanduíche de citocinas pró-inflamatórias em sobrenadante de culturas de esplenócitos de camundongos BALB/c. Ambos os extratos apresentaram flavonoides, derivados cinâmicos, esteróides e ácido elágico. O PBO mostrou atividade bacteriostática contra Acinetobacter baumannii (CIM = 250 µg.mL-¹) e atividade fungistática do PBA contra Trichophyton rubrum (CIM = 800 µg.mL-¹). Os extratos não apresentaram toxicidade para CMSP e células neoplásicas (IC50 > 100 µg.mL-¹). Além disso, o PBO a 100 µg.mL-¹ inibiu significativamente as citocinas IFN-γ e IL-17A (p = 0,03). Plectranthus barbatus é um candidato potencial para uso terapêutico devido à sua baixa toxicidade em células humanas saudáveis e exibe atividade de interesse médico como bacteriostática, fungistática e imunomoduladora.

Effects of Vitamin A and K3 on Immune Function and Intestinal Antioxidant Capacity of Aged Laying Hens

This study was conducted to investigate effects of vitamin A (VA) and vitamin K3 (VK3) on immune function and intestinal antioxidant capacity of aged laying hens. In a 3 × 3 factorial arrangement, the diets of 1080 Roman Pink laying hens (87 weeks old) was formulated with deficient, adequate and excess VA and VK3, including 0, 7000 and 14000 IU/kg VA and 0, 2.0 and 4.0 mg/kg VK3 for 8 weeks. Interactive effects between VA and VK3 were observed that VA and VK3 decreased the splenetic mRNA expression of inducible nitric oxide synthase (iNOS) and tumour necrosis factor α (TNF-α), but increased the plasma immunoglobulin G (IgG) content and jejunal mRNA expression of nuclear factor-like 2 (Nrf2). Hens fed adequate or excess VA had higher spleen index, mRNA expression of interleukin-10 (IL-10) in spleen, sIgA content, catalase (CAT), glutathione peroxidase and total dismutase (T-SOD) activity, and mRNA expression of polymeric immunoglobulin receptor (pIgR) in jejunum and lower mRNA expression of IL-1ß in jejunum and iNOS, TNF-α in spleen. Furthermore, adequate or excess VK3 significantly increased plasma IgG content, the CAT, T-SOD and total antioxidant capacity activities, up-regulated the mRNA expression of pIgR, Nrf2, SOD1 and CAT in jejunum and down-regulated the mRNA expression of iNOS and TNF-α in spleen.(AU)