To investigate the effect and mechanism of NK-1 Tachykinin receptor (NK-IR) antagonist on hypoxia induced hepatic injury, we established the hypoxic rat model. 30 male SD rats (weighing 240-300g) were randomly divided into 3 groups, control group, and experimental groups including the hypoxia group and the NK-1R antagonist group. The rats of experimental groups underwent hypoxia, among them the NK-1R antagonist group were those with interference of NK-1R antagonist by intraperitoneal injection. Hepatic injury was evaluated by pathological staining, hepatic function detection and hepatocellular apoptosis determination. Results showed hypoxia-induced hepatic injury in rats was established successfully. Edema,ballooning degeneration and spotty necrosis were found in livers in the experimental groups, among which the pathological injury in the hypoxia group was worse than that in the NK-1R antagonist group. Moreover,GGT and the rate of hepatocellular apoptosis in the NK-1R antagonist group were obviously lower than that in the hypoxia group (P<0.05). But no significant difference were found in ALT,AST and ALP between groups (P>0.05). These data indicate that Substance P possibly participate in the process of hypoxia-induced hepatic injury, and NK-1R antagonist could reduce hypoxia-induced hepatic injury.
Animals ; Apoptosis ; drug effects ; Hepatocytes ; pathology ; Hypoxia ; pathology ; physiopathology ; Liver ; physiopathology ; Liver Function Tests ; Male ; Neurokinin-1 Receptor Antagonists ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the influence of non-peptide neuro-kinase 1 (NK1) receptor antagonist L-703, 606 on the early tissue edema formation in rats with deep partial-thickness scald.

METHODSOne hundred and fifty-two SD rats were enrolled in the study and were randomly divided into normal control (NC, n = 8), scald control (SC, n = 48, with 20% TBSA deep partial thickness scald), L-703, 606 treatment (LT, n = 48, with 20% TBSA deep partial-thickness scald 20 minutes after caudal vein injection of 250 nmol/kg L-703, 606) and beta-aescin treatment (AT, n = 48, with 20% TBSA deep-partial-thickness scald 30 minutes after caudal vein injection of 1.8 mg/kg beta-aescin) groups. The rats were sacrificed at 1, 4, 8, 24, 48 and 72 post scald hours (PSHs), with 8 rats at each time point. The peri-wound tissue and jejunum samples were harvested for the detection of vascular permeability and tissue water content with modified Evans blue extravasation method and average water content assay.

RESULTSThe vascular permeability was significantly higher in the peri-wound tissue and jejunum in SC, LT an AT groups than that in NC group (P < 0.01) at 1 PSH, and it decreased gradually at 4 PSH. The vascular permeability in the peri-wound tissue in LT and AT group was significantly lower than that in SC group (P < 0.01), and that in LT group was markedly higher than that in AT group (P < 0.01) at 48 and 72 PSHs. The vascular permeability of jejunum tissue in LT group was lower than that in SC group within 24 PSH (P < 0.01), while that in AT group was lower than that in LT group at the early postscald stage (P < 0.01), but no obvious difference was found between the two groups after 72 PSH (P > 0.05). The change in the tissue water content was as follows: Dehydration was observed in peri-wound tissue in SC, LT and AT groups at 1 PSH. The tissue water content increased gradually thereafter and reached the peak at 8 and 24 PSH. Certain degree of dehydration was observed in jejunum tissue in SC, LT and AT groups at early postscald stage. The water content in jejunum tissue in LT group was evidently higher than that in SC and AT groups (P < 0.05 or 0.01), edema was evident at 8 PSH, and it became more obvious at 48 PSH, then it subsided gradually. Edema was less evident in LT group.

CONCLUSIONNonpeptide NK1-receptor antagonist L-703, 606 was able to mitigate the vascular permeability and reduce tissue water content in peri-wound and jejunal tissues.

Animals ; Burns ; metabolism ; Capillary Permeability ; Disease Models, Animal ; Edema ; drug therapy ; Neurokinin-1 Receptor Antagonists ; Quinuclidines ; therapeutic use ; Rats ; Rats, Sprague-Dawley

OBJECTIVE: : To investigate the effect of nicotinamide phosphoribosyltransferase (NAMPT) inhibitor FK866 on the migration of human non-small cell cancer A549 cells and related mechanism. METHODS: : The inhibition effect of FK866 on A549 cells was tested by MTT assay. A549 cells were treated with 1.0 and 10.0 nmol/L FK866, and the cell migration was evaluated by modified wound scratch assay. The mRNA expression of E-cadherin and vimentin was detected by real-time RT-PCR, and the expression of ERK1/2 and pERK1/2 was determined by Western blotting. RESULTS: : FK866 inhibited the proliferation of A549 cells in a time-and concentration-dependent manner; after treatment for 72 h, the IC of FK866 was 9.55 nmol/L. When 1.0 nmol/L or 10.0 nmol/L FK866 was continuously applied 48 h before and 48 h after a scratch was made in wound scratch assay, the migration of A549 cells was significantly inhibited. However, when the FK866 was applied only 48 h after the scratch, the migration of A549 cells was inhibited by 10.0 nmol/L but not by 1.0 nmol/L FK866. The mRNA expression of E-cadherin and vimentin, and the activated ERK1/2 were significantly increased after 1.0 nmol/L FK866 treatment for 72 h. The pretreatment with nicotinamide adenine dinucleotide (NAD) precursor nicotinamide mononucleotide(1.0 mmol/L) or ERK1/2 inhibitor U0126 (10.0 μmol/L) reversed the up-regulation of E-cadherin and vimentin expression induced by FK866. CONCLUSIONS s: Low concentration of FK866 decreases the migration of A549 cells through the inhibition of NAD level, activation of ERK1/2 and up-regulation of E-cadherin expression. However, it also up-regulates the expression of vimentin, indicating that it may have dual effects on the migration of tumor cells.
A549 Cells ; Cadherins ; genetics ; Cell Movement ; drug effects ; Gene Expression Regulation ; drug effects ; Humans ; Morpholines ; pharmacology ; Neurokinin-1 Receptor Antagonists ; pharmacology ; Nicotinamide Phosphoribosyltransferase ; antagonists & inhibitors ; Piperazines ; pharmacology ; Vimentin ; genetics

The aim of this study was to, from the point of neurogenic inflammation, explore the pathogenesis of colitis and to provide direct evidence for the neurogenic colitis hypothesis. Male Sprague-Dawley rats (180-220 g) anesthetized with chloral hydrate were intrathecally (ith) implanted with polyethylene-10 (PE-10) catheter to reach the spinal cord T₁₂-L₅ level. Substance P (SP) was ith injected once a day for 14 d. The disease active index (DAI) score was calculated by rat body weight and stool. The macroscopic and HE staining-microscopic pathologies of colon/spinal tissue were evaluated. By immunofluorescence staining, the protein expression of a pro-inflammatory cytokine, migration inhibitory factor (MIF), in colon tissue was detected and was semi-quantitatively analyzed. The results showed that in the colon tissue, inflammation was dose-dependently aggravated by ith SP 10 μ and 20 μ, whereas in the spinal tissue, only slight edema and congestion were seen in SP 20 μ group. The MIF protein of colon tissue was increased in ith SP 10 μ and 20 μ groups (P<0.05, P<0.01 as compared to normal saline group respectively), but in the spinal tissue, there was no obvious MIF protein expression either in SP groups or in normal saline group. Pretreatment with neurokinin-1 (NK₁) receptor antagonist ([D-Pro2, D-Trp7, 9] -SP, 22.4 μ, ith, 10 min before ith SP) prolonged the latency of DAI rising and reduced the DAI amplitude, as well as prevented the high MIF expression induced by ith SP. These results suggest that rat colitis can be induced by direct SP stimulation in lumbar spine via activating central NK₁ receptor; and that colonic MIF is possibly one of the inflammatory factors involved in this pathogenesis. These data provide a reasonable support to the hypothesis of colitis being a neurogenic inflammation. In addition, a potential clinical significance for the finding that higher concentration of spinal SP can induce colitis via NK₁ receptor is discussed.
Animals ; Colitis ; chemically induced ; Colon ; pathology ; Disease Models, Animal ; Inflammation ; pathology ; Injections, Spinal ; Intramolecular Oxidoreductases ; metabolism ; Macrophage Migration-Inhibitory Factors ; metabolism ; Male ; Neurokinin-1 Receptor Antagonists ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, Neurokinin-1 ; metabolism ; Spinal Cord ; pathology ; Substance P ; adverse effects

OBJECTIVETo study the changes in the migration of airway smooth muscle cells (ASMC) in asthmatic rats with airway remodeling and the effect of NK-1R inhibitor WIN62577 on the migration of ASMC.

METHODSSprague-Dawley rats were randomly assigned into two groups: airway remodeling induced by asthma and normal control. ASMC from rats with asthma and airway remodeling induced by ovalbumin (OVA) inhalation for 8 weeks were primary cultured and purified. Immunofluorescence and real-time PCR were used to measure the expression of NK-1R. With NK-1R inhibitor WIN62577 treatment, the changes in the migration of ASMC were measured by transwell chambers.

RESULTSNK-1R in ASMC was expressed mainly in the cytoplasm and cell membrane in the airway remodeling group, and the mRNA expression of NK-1R was higher than the normal control group (P<0.01). The number of the migrated ASMC in the airway remodeling group was significantly higher than that in the normal control group (P<0.01). Various concentrations (10-11 mol/L, 10-10 mol/L, 10-9 mol/L and 10-8 mol/L) of WIN62577 treatment decreased the number of the migrated ASMC (P<0.05).

CONCLUSIONSNK-1R may affect airway remodeling possibly through promoting the migration ability of ASMC in rats with asthma.

Airway Remodeling ; drug effects ; Androstenes ; pharmacology ; Animals ; Asthma ; pathology ; Benzimidazoles ; pharmacology ; Cell Movement ; drug effects ; Female ; Myocytes, Smooth Muscle ; drug effects ; physiology ; Neurokinin-1 Receptor Antagonists ; pharmacology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the role of substance P in the formation of hypertrophic scar.

METHODSDermal fibroblasts derived from human normal skin were cultured with substance P alone or together with selective non-peptide NK-1 tachykinin antagonist, L-703, 606 oxalate salt. The effect of substance P on proliferation of fibroblasts was measured by MTT assay. Furthermore, the TGF-beta 1 mRNA expression in the fibroblasts was determined by in situ hybridization and image analysis.

RESULTSSubstance P enhanced fibroblast proliferation dose-dependently, which showed the maximum rate when the concentration of substance P was 25 ng/ml or higher in the culture media. By 48 hours cultured with 25 ng/ml of substance P, the fibroblasts expressed TGF-beta 1 mRNA more significantly than the fibroblasts without substance P. The effects of substance P on both fibroblast proliferation and TGF-beta 1 mRNA expression could be antagonized by L-703, 606 oxalate salt.

CONCLUSIONThe results suggest that substance P may play an important role in phenotype changes of fibroblasts in skin scarring.

Cell Division ; Cells, Cultured ; Dermis ; cytology ; Fibroblasts ; cytology ; drug effects ; metabolism ; Gene Expression ; drug effects ; Neurokinin-1 Receptor Antagonists ; Quinuclidines ; pharmacology ; RNA, Messenger ; Substance P ; metabolism ; pharmacology ; Transforming Growth Factor beta ; genetics ; Transforming Growth Factor beta1 ; Up-Regulation