Adolescent ; Child ; Female ; Humans ; Male ; Nasal Mucosa ; metabolism ; Nasal Polyps ; metabolism ; Neuropeptides ; classification ; metabolism

OBJECTIVETo study the interaction among aquaporin1 (AQP), aquaporin2 (AQP2) and antisecretory factor( AF) , and their expression in the rat inner ear for furthur understanding of Meniere' s disease.

METHODSInner ear tissue section of six healthy male Sprague-Dawley rats was performed and Envision immunochemical staining was applied to detect the expression of AF, AQP1 and AQP2 in the rat inner ear. Vestibular and cochlear tissues of twenty healthy male Sprague-Dawley rats were dissected. Coimmunoprecipitation and Western Blot were used to specifically immunoprecipitate AF protein in the vestibular and cochlear tissues with monoclonal antibodies against AQP1 and polyclonal antibodies antibodies against AQP2 to detect the above precipitate with specific antibodies against AF.

RESULTS(1) AF was widely distributed in the inner ear, such as marginal cells of stria vascularis , five classes of spiral ligament fibrocyte , Reissner's membrane, basilar membrane, ampullar crest and so on with mild or moderate staining. In addition, round membrane was moderately or markedly stained. Positive immunostaining was found in the cochlear spiral ganglion, vestibular nerve and cochlear nerve. AQP1 was distributed in the intermediate cells in stria vascularis, type III fibrocyte of spiral ligament, basilar membrane and round membrane with moderate to marked degree of immunostaining intensity. AQP2 was mainly localized to the type II, IV, and V fibrocyte of spiral ligament, with moderate to marked degree of immunostaining intensity, round membrane was weakly stained. (2) No band was observed in the control and a single immunoreactive band of 60 000 was observed, which was equal to the molecular mass of AF.

CONCLUSIONS(1) AF, AQP1 and AQP2 have its individual specific localization in the rat inner ear, which was close to the parts of endolymph, so regulating water of the endolymph may be possible. (2) The range of localization of AF overlapped the distribution of AQP1 and AQP2. The results showed the existence of AF protein in the immunoprecipitate using co-immunoprecipitation combined with Western Blot. It suggested that the interaction between AQP1, AQP2 and AF might be possible.

Animals ; Aquaporin 1 ; metabolism ; Aquaporin 2 ; metabolism ; Cochlea ; metabolism ; Ear, Inner ; metabolism ; Male ; Neuropeptides ; metabolism ; Rats ; Rats, Sprague-Dawley

AIMTo investigate the changes and probable roles of adrenomedullin2/intermedin (AIDM2/IMD), a novel micromolecular bioactive peptide, in the lungs of rats with chronic hypoxic pulmonary hypertension.

METHODSTwenty male SD rats were randomly divided into normal control group (NC) and normobaric hypoxia group (4H). The protein levels of ADM and ADM2/IMD) in the plasma and lung were measured by radioimmunoassay and immunohistochemistry. The mRNA expressions of ADM, ADM2/IMD and their receptors C (RLR, RAMP1, RAMP2 and RAMP3 in the lung tissue were determined by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTS(1) The rat model of chronic pulmonary hypertension was confirmed by the increased mean pulmonary arterial pressure (mPAP) and weight ratio of right ventricle to left ventricle plus septum [RV/(LV + S)] in 4H group compared to NC group. (2) The concentrations of ADM in the plasma and lung homogenate of 4H group were 2.3 and 3.2 folds of NC group, respectively (all P < 0.01). The levels of ADM2/IMD were higher 89.6% and 45.0% in the plasma and lung homogenate of 4H group than those of NC group (respectively, P < 0.01, P < 0.05). (3) The mRNA expressions of ADM2/IMD and ADM in the lung of 4H group were up-regulated (respectively, P < 0.01, P < 0.05 vs. NC group). The expressions of CRLR and RAMP1 mRNAs were down-regulated (all P < 0.01 vs. NC group), while the levels of RAMP2 and RAMP3 mRNAs were no significant difference between the two groups. (4) The strong ADM2/IMD immunostaining was detected in the endothelial and adventitial cells of the rat pulmonary arteriole.

CONCLUSIONADM2/IMD, like its paralog ADM, might be closely related to the chronic hypoxic pulmonary hypertension in rats. The disorders of the gene expression and/or the synthesis and metabolism of ADM2/IMD and its receptor CRLR/RAMP1 possibly take part in the pathogenesis of chronic hypoxic pulmonary hypertension in rats.

Adrenomedullin ; metabolism ; Animals ; Hypertension, Pulmonary ; etiology ; metabolism ; Hypoxia ; complications ; metabolism ; Lung ; metabolism ; Male ; Neuropeptides ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVE[corrected] To characterize the insulinotropic action of hippocampal cholinergic neurostimulating peptide (HCNP) and analyze the role of type 3 muscarinic receptor (M(3)R) pathway in the action of HCNP.

METHODSINS-1 cells were incubated in routine RPMI 1640 medium (control group), RPMI 1640 supplemented with 50 pg/ml synthetic HCNP (HCNP group), or HCNP-containing medium with the addition of PMA 18 h prior to insulin release assay. The insulin levels in the medium was measured using radioimmunoassay following stimulation with different concentrations of glucose. Real-time quantitative PCR was used for detecting the gene expression of HCNP-pp, choline acetyltransferase (ChAT) and M(3)R in HCNP group and control group.

RESULTSAfter stimulation with different concentrations of glucose (5.6 and 16.7 mmol/L), HCNP group showed significantly higher insulin levels than the control and HCNP+ PMA groups. Compared with those in the control group, the mRNA levels of HCNP-pp, ChAT, and M(3)R were all lowered in HCNP group.

CONCLUSIONHCNP can promote insulin release in INS-1 cells by increasing ChAT activity and activating M(3)R, and this effect is inhibited by PMA.

Animals ; Cell Line ; Insulin ; secretion ; Neuropeptides ; pharmacology ; Rats ; Receptor, Muscarinic M3 ; metabolism

The current epidemic of obesity and its associated metabolic syndromes impose unprecedented challenges to our society. Despite intensive research on obesity pathogenesis, an effective therapeutic strategy to treat and cure obesity is still lacking. Exciting studies in last decades have established the importance of the leptin neural pathway in the hypothalamus in the regulation of body weight homeostasis. Important hypothalamic neuropeptides have been identified as critical neurotransmitters from leptin-sensitive neurons to mediate leptin action. Recent research advance has significantly expanded the list of neurotransmitters involved in body weight-regulating neural pathways, including fast-acting neurotransmitters, gamma-aminobutyric acid (GABA) and glutamate. Given the limited knowledge on the leptin neural pathway for body weight homeostasis, understanding the function of neurotransmitters released from key neurons for energy balance regulation is essential for delineating leptin neural pathway and eventually for designing effective therapeutic drugs against the obesity epidemic.
Animals ; Energy Metabolism ; Gene Expression ; Humans ; Hunger ; Hypothalamus ; metabolism ; physiology ; Leptin ; metabolism ; physiology ; Neural Pathways ; metabolism ; Neuropeptides ; genetics ; metabolism ; Obesity ; metabolism

OBJECTIVETo investigate the changes of encephalic contents of somatostatin (SS) and arginine vasopressin (AVP) in rats with vascular dementia.

METHODSTwenty-four male Sprague-Dawley rats were randomly divided into three groups: vascular dementia group (VDMG), sham operation group (SOG) and control group (CG). The vascular dementia model was established by permanent bilateral vertebral artery occlusion through electric coagulation and shut-off the bilateral carotid arteries. The remember behavior of animals was tested and the contents of SS and AVP in various encephalic region (frontal cortex, temporal lobe, hippocampus, cerebral ganglion and corpora striatum) were determined with radioimmunoassay.

RESULTDuring the 15-day-long remembering test, the frequency of making mistakes in the VDMG was higher remarkably than that in SOG and CG (P<0.05); and the relative contents of SS were decreased in frontal area cortex, temporal lobe, hippocampus, cerebral ganglion and corpora striatum (P<0.01), while decrease of AVP contents was only detected in temporal lobe and corpora striatum (P<0.05).

CONCLUSIONThe disturbance of learning and memory function might be associated with SS and AVP after multiple cerebral infarction in the animals.

Animals ; Arginine Vasopressin ; metabolism ; Brain ; metabolism ; Dementia, Vascular ; metabolism ; physiopathology ; Male ; Memory ; physiology ; Neuropeptides ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Somatostatin ; metabolism

To understand the etiology of metabolic disorders, including obesity and type II diabetes, it is essential to gain better insight into how stored and available energy sources are monitored by the central nervous system. In particular, a comprehension of the fine cellular interplay and intracellular mechanisms that enable appropriate hypothalamic and consequent endocrine and behavioral responses to both circulating hormonal and nutrient signals remains elusive. Recent data, including those from our laboratories, raised the notion that reactive oxygen species (ROS) generation is not merely a by-product of substrate oxidation, but it plays a crucial role in modulating cellular responses involved in the regulation of energy metabolism. These review summarizes the published recent data on the effect of ROS levels in the regulation of neuronal function, including that of hypothalamic melanocortin neurons, pro-opiomelanocortin and neuropeptide Y-/agouti related peptide-neurons, in the modulation of food intake.
Central Nervous System ; Comprehension ; Eating ; Energy Metabolism ; Hypothalamus ; Neurons ; Neuropeptides ; Obesity ; Peroxisomes ; Pro-Opiomelanocortin ; Reactive Oxygen Species

OBJECTIVETo detect the expression of protein 4.1 family members in mouse melanoma cell lines and evaluate their effect on cell proliferation.

METHODSPCR and Western blot were used to detected to the expression of protein 4.1 family members (4.1R, 4.1B, 4.1G, and 4.1N) at the mRNA and protein levels in B16 and B16-F10 cell lines. The expression plasmid vector pEGFP-N1-EPB41L3 carrying 4.1B gene sequence amplified from genomic RNA of mouse embryo fibroblasts was constructed and transiently transfected into mouse melanoma cells. The change in cell proliferation was assessed using MTT assay.

RESULTSThe mRNA and protein expressions of all the protein 4.1 family members, with the exception of 4.1B, were detected in both B16 and B16-F10 cells. Transfection of cells with the eukaryotic expression vector pEGFP-N1-EPB41L3 markedly inhibited cell proliferation as compared with the non-transfected cells.

CONCLUSIONThe eukaryotic expression vector carrying EPB41L3 sequence is capable of inhibiting the proliferation of mouse melanoma B16 and B16-F10 cells.

Animals ; Cell Line, Tumor ; Cell Proliferation ; Cytoskeletal Proteins ; metabolism ; Genetic Vectors ; Melanoma, Experimental ; metabolism ; Membrane Proteins ; metabolism ; Mice ; Microfilament Proteins ; Neuropeptides ; metabolism ; Plasmids ; Transfection

Animals ; Disease Models, Animal ; Hypoglycemia ; chemically induced ; metabolism ; Hypothalamus ; drug effects ; metabolism ; Insulin ; pharmacology ; Intracellular Signaling Peptides and Proteins ; metabolism ; Male ; Neuropeptides ; metabolism ; Orexins ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo analyze the level of the oxytocin (OT) and the expression of oxytocin receptor (OTR) in males with idiopathic infertility.

METHODSSixty-five infertile males aged 20 -45 years were divided according to their semen parameters into an idiopathic oligozoospermia group (OG, n = 20), an idiopathic asthenozoospermia group (AG, n = 25), and an idiopathic oligoasthenozoospermia group (OAG, n = 20). Another twenty 20-45 years old healthy male volunteers with a natural childbearing history were included in the control group (CG). All the subjects were detected for the contents of luteinizing hormone (LH), follicle stimulating hormone (FSH) , testosterone (T) and OT, and analyzed for the expression of OTR by sequencing the functional region of the OTR promoter (OTRP), OTR-mRNA, and OTR-COOH terminus. The gene sequences were compared using DNASTAR-MegAlign, Western blot values changed into enumeration data, and all the data analyzed by one-way ANOVA and Dunnette's multiple range t-test.

RESULTSA significantly lower content of OT was observed in CG ( [79.30 +/- 3.83] pg/ml) than in OG ([118.53 +/- 7.69] pg/ml, AG ([108.81 +/- 5.66] pg/ml) and OAG ([103.71 +/- 4.54] pg/ml) (F(0.05/2[2,82]) = 8.29, P < 0.01). The content of LH was significantly lower in AG ([4.26 +/- 0.31] IU/L) and OAG ([4.55 +/- 0.40] IU/L) than in OG ([6.77 +/- 0.57] IU/L) and CG ([7.19 +/- 0.50] IU/L) (F(0.05/2 [2,82]) = 11.64, P < 0.01), and so was the content of FSH in AG ( [5.02 + 0.39] IU/L) than in CG ([8.91 +/- 0.91] IU/L), OG ([11.86 +/- 1.76] IU/L) and OAG ([8.82 +/- 1.03] IU/L) (F(0.05/[2,82]) = 7.22, P < 0.01). There were no significant differences in the T content among the four groups (F(0.05/2[2,82] = 0.42, P = 0.739). No evident gene mutation was found in OTRP and OTR-mRNA gene sequencing. Human OTRs in the lymphocytes were monomers and oligomers, mostly tetramers and hexamers. There were obviously more monomers in AG (0.41 +/- 0.03) and OAG (0.13 +/- 0.01) than in OG (0.05 +/- 0.004) and CG (0.05 +/- 0.003) (F(0.05/2[2,82]) = 115.50, P < 0.01), while the number of oligomers was markedly decreased in 20% of the cases in AG.

CONCLUSIONSignificant differences in the content of OT and expression of OTR between fertile and infertile men suggested an association of OT with male infertility. The decreased expression of OTR oligomers and increased expression of monomers may be related to idiopathic asthenozoospermia, which has provided a new insight into the pathogenesis and treatment of male infertility.

Adult ; Case-Control Studies ; Humans ; Infertility, Male ; metabolism ; pathology ; Male ; Middle Aged ; Neuropeptides ; metabolism ; Oxytocin ; metabolism ; Receptors, Oxytocin ; metabolism ; Young Adult