Although little is known on the function of gammadelta T lymphocytes, there is increasing evidence that gammadelta T lymphocytes are early responders and modulators of immune responses against pathogens and cytokines such as IL-2, IL-7, IL-15 and TNF-alpha. To study the role TNF-alpha on gammadelta T lymphocytes, we cloned bovine TNF-alpha. Sequence analysis revealed that a new allele form of bovine TNF-alpha was cloned which has 3 additional nucleotide sequences as well as 3 nucleotide substitutions compared with previously reported bovine TNF-alpha. Further studies are needed to document the functional significance of a new allele form of TNF-alpha in cattle.
*Alleles ; Amino Acid Sequence ; Animals ; Base Sequence ; Cattle ; *Cloning, Molecular ; Gene Library ; Molecular Sequence Data ; Receptors, Antigen, T-Cell, gamma-delta/*physiology ; Sequence Alignment/veterinary ; T-Lymphocytes/immunology/*physiology ; Tumor Necrosis Factor-alpha/chemistry/*genetics/physiology

OBJECTIVETo observe the effect of volatile oil of Schizonepetae Herba (VOSH), and its essential components-menthone and pulegone against anti-influenza virus A/PR/8/34 (H1N1) in vivo and in vitro, as well as the signaling mechanism of its toll-like receptor/interferon (TLR/IFN).

METHODThe lung-adapted PR-8 virus model was prepared in mice. They were administered with preventive and therapeutic drugs, and the hemagglutination titer of model animals was determined to evaluate in vivo effect against H1N1. ELISA test was conducted to observe the effect on IFN-alpha, IFN-beta, IL-2, IL-6 and TNF-alpha in serum, as well as IFN-beta secretion in H1N1 infected MDCK supernatant. Real-time RT-PCR was employed to observe the expression levels of IRAK4 and TLR3 mRNA.

RESULTThe in vivo experiment shows that the hemagglutination titer was significantly decreased when the mice were treated with VOSH (0.266 mg x kg(-1)), menthone(0.5 mg x kg(-1)) and pulegone (0.19 mg x kg(-1)) in therapeutic way; VOSH (0.226 mg x kg(-1)) had a significant effect on increasing serum levels of IFN-alpha, IL-2; Methone (0.5 mg x kg(-1)) had a significant effect on increasing serum levels of IFN-beta; Methone (0.5 mg x kg(-1)) and pulegone (0.19 mg x kg(-1)) had a significant effect on decreasing serum levels of IL-6; VOSH (0.452, 0.226 mg x kg(-1)) and pulegone (0.19 mg x kg(-1)) had a significant effect on decreasing serum levels TNF-alpha. The in vitro experiment showed that the expression levels of IRAK4 mRNA and IFN-beta were significantly increased in VOHS (0.1 g x L(-1)) and pulegone (0.1 g x L(-1)) groups; and the menthone (0.25 g x L(-1)) group showed a significant rise in the expression levels of IRAK4 mRNA, but a notable decline in TLR3 mRNA.

CONCLUSIONThe administration with VOSH, methone and pulegone in therapeutic way can significantly decrease the hemagglutination titer, which demonstrates the anti-virus effect of the administration in therapeutic way, but no notable efficacy of the administration in preventive way. The in vivo anti-virus mechanism is related to regulation of IFN-alpha, IFN-beta and IL-2.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Humans ; Influenza A Virus, H1N1 Subtype ; drug effects ; physiology ; Influenza, Human ; drug therapy ; genetics ; immunology ; virology ; Interferon-alpha ; genetics ; immunology ; Interleukin-1 Receptor-Associated Kinases ; Interleukin-2 ; genetics ; immunology ; Interleukin-6 ; genetics ; immunology ; Lamiaceae ; chemistry ; Male ; Mice ; Oils, Volatile ; pharmacology ; Tumor Necrosis Factor-alpha ; genetics ; immunology

OBJECTIVETo investigate the effect of shenfu injection on canine with cardiogenic shock and the possible mechanism.

METHODCardiogenic shock model of canine was established by ligating left anterior descending (LAD) of coronary artery. The 15 canines with cardiogenic shock were randomly divided in to glucose injection group, shenfu injection group and sham-operated group. The hemodynamics parameters were monitored. Plasma TNF-alpha and IL-1beta levels were measured by radioimmunoassay. Expression of TNF-alpha mRNA and IL-1beta mRNA in myocardium were detected by RT-PCR.

RESULTFollowing cardiogenic shock, the mean artery pressure (MAP), left ventricular systolic pressure (LVSP), ventricular pressure rise ratio during systolic period (+ dp/dt(max)), and ventricular pressure decay ratio during diastolic period (- dp/dt(max)) decreased significantly; the plasma TNF-alpha and IL-1beta levels and the expression of TNF-a mRNA and IL-1beta mRNA in myocardium increased significantly. In shenfu injection group, MAP, LVSP and +/- dp/dt(max) increased significantly and plasma TNF-alpha and IL-1beta levels decreased significantly. In glucose injection group, MAP, LVSP, +/- dp/dt(max) and plasma TNF-alpha and IL-1beta levels had not changed significantly. The expression of TNF-alpha mRNA and IL-1beta mRNA in myocardium were significantly lower in shenfu injection group than those in glucose injection group.

CONCLUSIONShenfu injection probably can decrease over-exprssion of TNF-alpha mRNA and IL-1beta mRNA on transcription platform. Shenfu injection counteract cardiogenic shock, relieve myocardium damage and improve hemodynamics through inhibiting overproduction of TNF-alpha and IL-1beta.

Aconitum ; chemistry ; Animals ; Cytokines ; biosynthesis ; blood ; genetics ; Dogs ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Female ; Hemodynamics ; drug effects ; physiology ; Injections, Intravenous ; Interleukin-1beta ; biosynthesis ; blood ; genetics ; Male ; Myocardium ; metabolism ; Panax ; chemistry ; Phytotherapy ; Plants, Medicinal ; chemistry ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Reverse Transcriptase Polymerase Chain Reaction ; Shock, Cardiogenic ; physiopathology ; prevention & control ; Tumor Necrosis Factor-alpha ; biosynthesis ; blood ; genetics

OBJECTIVETo explore the effects of traditional Tibetan medicine, Fructus Lonicerae microphyllae (FLM) on phagecytosis and cytokines production of murine macrophages.

METHODThe phagecytosis of murine macrophages was analyzed by neutral red phagecytosis assay. The activities of IL-1 and TNF-alpha were measured by biological methods. The mRNA of TNF-alpha and INF-gamma expressed by macrophages was detected by RT-PCR.

RESULTThe phagecytosis of murine macrophages was significantly enhanced by FLM at a concentration from 1 microg x mL(-1) to 100 microg x mL(-1) and the secretions of IL-1, and TNF-alpha from macrophages were markedly induced by FLM. Meanwhile, FLM also increased the expression of TNF-alpha mRNA and INF-gamma mRHA from macrophages in vitro.

CONCLUSIONFLM could promote phagecytosis and cytokines production of murine macrophages.

Animals ; Cell Line ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Fibroblasts ; cytology ; Fruit ; chemistry ; Interferon-gamma ; biosynthesis ; genetics ; Interleukin-1 ; secretion ; Lonicera ; chemistry ; Macrophages, Peritoneal ; metabolism ; physiology ; Mice ; Mice, Inbred C57BL ; Mice, Inbred ICR ; Phagocytosis ; drug effects ; Plants, Medicinal ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics

OBJECTIVETo discuss the role of alveolar macrophage activation in rats with acute necrotizing pancreatitis (ANP) associated with lung injury.

METHODS30 adult Sprague-Dawley rats were randomly divided into five groups (n = 6): normal control group, one-hour group, three-hour group, six-hour group and twelve-hour group after ANP induction. ANP was induced by intraductal administration of 3% sodium taurocholate, while the normal control received an infusion of physiological saline. Alveolar macrophages were harvested by bronchoalveolar lavage. The protein content of lavage fluids, the myeloperoxidase of lung tissue (MPO), and tumor necrosis factor alpha (TNFalpha) and nitric oxide (NO) secreted by alveolar macrophages were examined. The expression of TNFalpha mRNA and inducible nitric oxide synthase (iNOS) mRNA was measured with reverse transcription-polymerase chain reaction technique. Histology of the lung and pancreas was scored in a blinded fashion.

RESULTSLung injury was gradually aggravated with disease progression. The level of myeloperoxidase of lung tissue and protein content of bronchoalveolar lavage fluids increased progressively and reached the peak at 12 hour [(10.78 +/- 0.58) U/g for MPO and (2 011.0 +/- 105.5) micro g/ml for protein respectively]. TNFalpha and NO secreted by alveolar macrophages were gradually elevated and peaked on the sixth hour, the maximums were (1 624.2 +/- 149.2) pg/ml and (88.8 +/- 6.5) micro mol/L respectively, but decreased on the twelfth hour. The expression of TNFalpha mRNA and iNOS mRNA was similar with the changes of TNFalpha and NO, upregulated after induction of acute necrotizing pancreatitis and reached their peaks on the sixth hour, then downregulated on the twelfth hour. All the parameters of ANP groups compared to control group were statistical significant (P < 0.05). The histology scores demonstrated an increasing damage of the lung. The expression of TNFalpha mRNA and iNOS mRNA is closely related to lung injury (r = 0.67 for TNFalpha mRNA and r = 0.64 for iNOS mRNA respectively, P < 0.01).

CONCLUSIONThe activation of alveolar macrophage may play an important role in lung injury associated with acute necrotizing pancreatitis.

Animals ; Bronchoalveolar Lavage Fluid ; chemistry ; Female ; Lung ; pathology ; Macrophage Activation ; Macrophages, Alveolar ; physiology ; Male ; Nitric Oxide ; Nitric Oxide Synthase ; genetics ; Nitric Oxide Synthase Type II ; Pancreatitis, Acute Necrotizing ; complications ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Respiratory Distress Syndrome, Adult ; etiology ; Tumor Necrosis Factor-alpha ; genetics

Matrix metalloproteinase-9 (MMP-9) secreted from macrophages plays an important role in tissue destruction and inflammation through degradation of matrix proteins and proteolytic activation of cytokines/chemokines. Whereas the MEK-ERK and PI3K-Akt pathways up-regulate MMP-9 expression, regulation of MMP-9 by JNK remains controversial. Presently, we aimed to determine the role of JNK in MMP-9 regulation in Raw 264.7 cells. Inhibition of JNK by the JNK inhibitor SP600125 induced MMP-9 in the absence of serum and suppressed the expression of TNF-alpha, IL-6 and cyclooxygenase-2 in LPS-treated Raw 264.7 cells. In a knockdown experiment with small interfering RNA, suppression of JNK1 induced MMP-9 expression. Interestingly, mouse serum suppressed SP600125-mediated MMP-9 induction, similar to IFN-gamma. However, the inhibitory activity of mouse serum was not affected by pyridone 6, which inhibits Janus kinase downstream to IFN-gamma. In addition to mouse serum, conditioned media of Raw 264.7 cells contained the inhibitory factor(s) larger than 10 kDa, which suppressed SP600125- or LPS-induced MMP-9 expression. Taken together, these data suggest that JNK1 suppresses MMP-9 expression in the absence of serum. In addition, the inhibitory factor(s) present in serum or secreted from macrophages may negatively control MMP-9 expression.
Animals ; Anthracenes/metabolism ; Cell Line ; Culture Media, Conditioned/*chemistry ; Enzyme Activation ; Enzyme Induction ; Enzyme Inhibitors/metabolism ; Extracellular Signal-Regulated MAP Kinases/genetics/metabolism ; *Gene Expression Regulation, Enzymologic ; MAP Kinase Signaling System/physiology ; Macrophages/cytology/*metabolism ; Matrix Metalloproteinase 9/genetics/*metabolism ; Mice ; Mitogen-Activated Protein Kinase 8/genetics/*metabolism ; NF-kappa B/genetics/metabolism ; Proto-Oncogene Proteins c-akt/genetics/metabolism ; Tumor Necrosis Factor-alpha/metabolism ; p38 Mitogen-Activated Protein Kinases/genetics/metabolism