RESUMO
The objective of this study was to investigate the relationship between PI3K/mTOR/RhoA signaling regulated cytoskeletal rearrangements and phagocytic capacity of macrophages. RAW264.7 macrophages were divided into four groups; blank control, negative control, PI3K-RNAi, and mTOR-RNAi. The cytoskeletal changes in the macrophages were observed. Furthermore, the phagocytic capacity of macrophages against Escherichia coli is reported as mean fluorescence intensity (MFI) and percent phagocytosis. Transfection yielded 82.1 and 81.5% gene-silencing efficiencies against PI3K and mTOR, respectively. The PI3K-RNAi group had lower mRNA and protein expression levels of PI3K, mTOR, and RhoA than the blank and negative control groups (Ð <0.01). The mTOR-RNAi group had lower mRNA and protein levels of mTOR and RhoA than the blank and the negative control groups (Ð <0.01). Macrophages in the PI3K-RNAi group exhibited stiff and inflexible morphology with short, disorganized filopodia and reduced number of stress fibers. Macrophages in the mTOR-RNAi group displayed pronounced cellular deformations with long, dense filopodia and an increased number of stress fibers. The PI3K-RNAi group exhibited lower MFI and percent phagocytosis than blank and negative control groups, whereas the mTOR-RNAi group displayed higher MFI and percent phagocytosis than the blank and negative controls (Ð <0.01). Before and after transfection, the mRNA and protein levels of PI3K were both positively correlated with mTOR and RhoA (Ð <0.05), but the mRNA and protein levels of mTOR were negatively correlated with those of RhoA (Ð <0.05). Changes in the phagocytic capacity of macrophages were associated with cytoskeletal rearrangements and were regulated by the PI3K/mTOR/RhoA signaling pathway.
Assuntos
Citoesqueleto/metabolismo , Macrófagos/metabolismo , Fagocitose/fisiologia , Fosfatidilinositol 3-Quinases/metabolismo , Serina-Treonina Quinases TOR/metabolismo , Proteína rhoA de Ligação ao GTP/metabolismo , Animais , Western Blotting , Inativação Gênica , Vetores Genéticos , Humanos , Camundongos , Células RAW 264.7 , Interferência de RNA , Reação em Cadeia da Polimerase em Tempo Real , Transdução de Sinais , TransfecçãoRESUMO
The objective of this study was to investigate the relationship between PI3K/mTOR/RhoA signaling regulated cytoskeletal rearrangements and phagocytic capacity of macrophages. RAW264.7 macrophages were divided into four groups; blank control, negative control, PI3K-RNAi, and mTOR-RNAi. The cytoskeletal changes in the macrophages were observed. Furthermore, the phagocytic capacity of macrophages against Escherichia coli is reported as mean fluorescence intensity (MFI) and percent phagocytosis. Transfection yielded 82.1 and 81.5% gene-silencing efficiencies against PI3K and mTOR, respectively. The PI3K-RNAi group had lower mRNA and protein expression levels of PI3K, mTOR, and RhoA than the blank and negative control groups (Р<0.01). The mTOR-RNAi group had lower mRNA and protein levels of mTOR and RhoA than the blank and the negative control groups (Р<0.01). Macrophages in the PI3K-RNAi group exhibited stiff and inflexible morphology with short, disorganized filopodia and reduced number of stress fibers. Macrophages in the mTOR-RNAi group displayed pronounced cellular deformations with long, dense filopodia and an increased number of stress fibers. The PI3K-RNAi group exhibited lower MFI and percent phagocytosis than blank and negative control groups, whereas the mTOR-RNAi group displayed higher MFI and percent phagocytosis than the blank and negative controls (Р<0.01). Before and after transfection, the mRNA and protein levels of PI3K were both positively correlated with mTOR and RhoA (Р<0.05), but the mRNA and protein levels of mTOR were negatively correlated with those of RhoA (Р<0.05). Changes in the phagocytic capacity of macrophages were associated with cytoskeletal rearrangements and were regulated by the PI3K/mTOR/RhoA signaling pathway.
Assuntos
Humanos , Animais , Ratos , Fagocitose/fisiologia , Citoesqueleto/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Proteína rhoA de Ligação ao GTP/metabolismo , Serina-Treonina Quinases TOR/metabolismo , Macrófagos/metabolismo , Transfecção , Transdução de Sinais , Western Blotting , Inativação Gênica , Interferência de RNA , Reação em Cadeia da Polimerase em Tempo Real , Células RAW 264.7 , Vetores GenéticosRESUMO
An experiment was conducted to investigate the effects of light intensity on growth, anti-stress ability, and immune function of yellow feathered broilers. A total of 480 one-day-old male Lingnan yellow feathered broilers were randomly allocated to 4 treatments based on light intensity (1, 5, 20 and 80 lx) with 8 replicates of 15 chicks each. The experiment lasted for 63 days. Compared with those under high light intensity, broilers exposed to low light intensity had higher (p<0.05) total antioxidant capacity (T-AOC), glutathione peroxidase (GSH-Px), a-Naphthylacetate esterase (ANAE+), antibody titer, but lower (p<0.05) malonaldehyde (MDA) levels and heterophil/lymphocyte ratio (H/L). There was a linear effect for T-AOC(p=0.002), GSH-Px(p≤0.047), MDA (p=0.003), H/L(p≤0.014), ANAE+ (p≤0.044), and antibody titer (p≤0.021) with T-AOC, GSH-Px, ANAE+, and antibody titer increased significantly as light intensity decreased, whereas MDA and H/L were decreased with the decrease in light intensity. These results suggested that broilers under low light intensity could have similar performance, better anti-stress ability, stronger immune function, and more efficient in energy usage as compared with those exposed to high light intensity environment.(AU)
Assuntos
Animais , Luz/efeitos adversos , Teste de Esforço/efeitos adversos , Aves Domésticas/anormalidadesRESUMO
An experiment was conducted to investigate the effects of light intensity on growth, anti-stress ability, and immune function of yellow feathered broilers. A total of 480 one-day-old male Lingnan yellow feathered broilers were randomly allocated to 4 treatments based on light intensity (1, 5, 20 and 80 lx) with 8 replicates of 15 chicks each. The experiment lasted for 63 days. Compared with those under high light intensity, broilers exposed to low light intensity had higher (p<0.05) total antioxidant capacity (T-AOC), glutathione peroxidase (GSH-Px), a-Naphthylacetate esterase (ANAE+), antibody titer, but lower (p<0.05) malonaldehyde (MDA) levels and heterophil/lymphocyte ratio (H/L). There was a linear effect for T-AOC(p=0.002), GSH-Px(p≤0.047), MDA (p=0.003), H/L(p≤0.014), ANAE+ (p≤0.044), and antibody titer (p≤0.021) with T-AOC, GSH-Px, ANAE+, and antibody titer increased significantly as light intensity decreased, whereas MDA and H/L were decreased with the decrease in light intensity. These results suggested that broilers under low light intensity could have similar performance, better anti-stress ability, stronger immune function, and more efficient in energy usage as compared with those exposed to high light intensity environment.
Assuntos
Animais , Aves Domésticas/anormalidades , Luz/efeitos adversos , Teste de Esforço/efeitos adversosRESUMO
Interleukin-18 (IL-18), an important proinflammatory cytokine, has been reported to play a potential pathological role in rheumatoid arthritis (RA). Results from previous studies on the association between IL-18 polymorphisms and RA are conflicting. To clarify this, an updated meta-analysis of all available studies on IL-18 polymorphisms and RA was conducted. Eligible articles were identified by searching databases, including PubMed, Ovid, Cochrane Library, EMBASE, and China Knowledge Resource Integrated Database, for the period up to May 1, 2015. The pooled odds ratios (ORs) with 95% confidence intervals (95%CIs) were used to assess the strength of association in the homozygote, heterozygote, dominant, recessive, and additive models. The software STATA (Version 13.0) was used for statistical analysis. Finally, 14 articles were included in the present meta-analysis. The IL-18 -607C/A polymorphism showed pooled ORs and 95%CIs for the homozygote model (AA vs CC: OR = 0.598; 95%CI = 0.395-0.907), and the association between the IL-18 -137G/C polymorphism and RA showed pooled ORs and 95%CIs for the homozygote (CC vs GG: OR = 0.699; 95%CI = 0.364-1.342) and heterozygote (CG vs GG: OR = 0.924; 95%CI = 0.803-1.064) models. In summary, the current meta-analysis, which was based on the most current studies, showed that the -607A/C, -920C/T, and -105A/C polymorphisms in IL-18 were significantly associated with increased RA risk. However, the -137C/G polymorphism was not associated with RA risk under any genetic model. More evidence is needed to support or deny such a conclusion.
Assuntos
Artrite Reumatoide/genética , Predisposição Genética para Doença , Interleucina-18/genética , Polimorfismo de Nucleotídeo Único , Povo Asiático/genética , Humanos , Razão de Chances , População Branca/genéticaRESUMO
Smith-Magenis syndrome (SMS) is a rare syndrome with multiple congenital malformations, including development and mental retardation, behavioral problems and a distinct facial appearance. SMS is caused by haploinsufficiency of RAI1 (deletion or mutation of RAI1). We describe an eight-year-old female Chinese patient with multiple malformations, congenital heart defect, mental retardation, and behavioral problems (self hugging, sleeping disturbance). High-resolution genome wide single nucleotide polymorphism array revealed a 3.7-Mb deletion in chromosome region 17p11.2. This chromosome region contains RAI1, a critical gene involved in SMS. To the best of our knowledge, this is the first report of an SMS patient in mainland China.
Assuntos
Deleção Cromossômica , Cromossomos Humanos Par 17/genética , Cardiopatias Congênitas/genética , Deficiência Intelectual/complicações , Deficiência Intelectual/genética , Síndrome de Smith-Magenis/complicações , Síndrome de Smith-Magenis/genética , Criança , Pré-Escolar , China , Fácies , Feminino , Cardiopatias Congênitas/complicações , Humanos , Recém-Nascido , Polimorfismo de Nucleotídeo Único/genéticaRESUMO
Insect olfactory perception involves many aspects of insect life, and can directly or indirectly evoke either individual or group behaviors. Insect olfactory receptors and odorant-binding proteins (OBPs) are considered to be crucial to insect-specific and -sensitive olfaction. Although the mechanisms of interaction between OBPs or OBP/ligand complex with olfactory receptors are still not well understood, it has been shown that many OBPs contribute to insect olfactory perception at various levels. Some of these are numerous and divergent members in OBP family; expression in the olfactory organ at high concentration; a variety of combinational patterns between different OBPs and ligands, but exclusive affinity for one OBP to specific binding ligands; complicated interactions between OBP/ligand complex and transmembrane proteins (olfactory receptors or sensory neuron membrane proteins). First, we review OBPs' ligand-binding property based on OBP structural research and ligand-binding test; then, we review current progress around the points cited above to show the role of such proteins in insect olfactory signal transmission; finally, we discuss applications based on insect OBP research.
Assuntos
Insetos/fisiologia , Receptores Odorantes/genética , Olfato/fisiologia , Animais , Feminino , Ligantes , Masculino , Percepção Olfatória/fisiologia , Neurônios Receptores Olfatórios/fisiologia , Feromônios/fisiologia , Receptores Odorantes/metabolismo , Transdução de Sinais/fisiologiaRESUMO
Inositol 1,4,5-triphosphate (Ins(1,4,5)P3) is a second messenger that regulates Ca2+ channels in many important cell signalling pathways. In sea urchin sperm the outer investment of the egg triggers the acrosome reaction (AR) that involves Ins(1,4,5)P3 production and the opening of two Ca2+ channels. Here we have sought to identify a high-affinity Ins(1,4,5)P3 receptor in Strongylocentrotus purpuratus sperm. An Ins(1,4,5)P3 binding component was affinity-purified 12-fold from sperm extracts. It displayed similar characteristics to the Ins(1,4,5)P3 receptor from other sources: pH-dependent high affinity for Ins(1,4,5)P3 (KD = 261 nM), a tau1/2 of association and dissociation of 50 and 40 s, respectively, specificity (IC50 > 5 microM for Ins(1)P1, Ins(1,4)P2 and Ins(1,3,4,5)P4), and pharmacological sensitivity (10 and 100 microg heparin/ml inhibited 75% and 100% binding respectively). An antibody against the carboxy-terminal of the type I Ins(1,4,5)P3 receptor of somatic cells recognised a plasma membrane component in the sperm head and less intensely in the flagella. This antibody also recognised a 240 kDa band from isolated head plasma membranes, and weakly in flagellar membrane. This IP3 receptor-like protein may mediate the sustained uptake of Ca2+ through the second Ca2+ channel opened during the AR.