To study the expression and distribution of neuropeptide Y (NPY) and long leptin receptor (OB-Rb) in the gastrointestinal tract of giant panda, samples of three animals were collected from the key laboratory for reproduction and conservation genetics of endangered wildlife of Sichuan province, China conservation and research center for the giant panda. Paraffin sections of giant panda gastrointestinal tissue samples were observed using hematoxylin-eosin staining (HE) and strept actividin-biotin complex immunohistochemical staining (IHC). The results show that the intestinal histology of three pandas was normal and no pathological changes, and there were rich single-cell and multi-cell mucous glands, long intestinal villi and thick muscularis mucosa and muscle layer. Positive cells expressing NPY and OB-Rb were widely detected in the gastrointestinal tract by IHC methods. NPY positive nerve fibers and neuronal cell were widely distributed in submucosal plexus and myenteric plexus, especially in the former. They were arranged beaded or point-like shape. NPY positive cells were observed in the shape of ellipse and polygon and mainly located in the mucous layer and intestinal glands. OB-Rb positive cells were mainly distributed in the mucous layer and the laminae propria, especially the latter. These results confirmed that NPY and OB-Rb are widely distributed in the gut of the giant panda, which provide strong reference for the research between growth and development, digestion and absorption, and immune function.
Animals ; China ; Intestines ; metabolism ; Neuropeptide Y ; genetics ; metabolism ; Receptors, Leptin ; genetics ; metabolism ; Ursidae ; genetics ; metabolism

Neuropeptide Y (NPY) is one of the most important orexigenic agents in central regulation of feeding behavior, body weight and energy homeostasis in domestic chickens. To examine differences in the hypothalamic NPY between layer-type and meat-type of chickens, which are two divergent kinds of the domestic chickens in feeding behavior and body weight, we detected mRNA levels of NPY in hypothalamic infundibular nucleus (IN), paraventricular nucleus (PVN) and lateral hypothalamic area (LHA) of these two types of chickens using one-step real time RT-PCR. The meat-type chicken had more food daily (about 1.7 folds) and greater body weights (about 1.5 folds) and brain weights than the layer-type chicken at the age of 14 d. In the meat-type of chicken, NPY mRNA levels of the IN and PVN were significantly greater than those of the LHA, and were not significantly different between the IN and PVN. However, in the layer-type of chicken, NPY mRNA levels were significantly greater in the IN than those in the LHA and PVN, and were not significantly different between the PVN and LHA. In all these hypothalamic regions, the layer-type of chicken had significantly higher NPY mRNA levels than the meat-type chicken did. These results suggest the expression of NPY in the hypothalamus has a type-dependent pattern in domestic chickens.
Animals ; Body Weight ; Chickens ; classification ; metabolism ; Hypothalamus ; metabolism ; Male ; Meat ; Neuropeptide Y ; genetics ; RNA, Messenger ; analysis

OBJECTIVEBenzodiazepines (BDZ) have many effects on various kinds of epilepsies, but long-term treatment with BDZ often leads to drug tolerance. This study aimed to seek drugs which can reverse the tolerance of flurazepam (FZP), and to explore the role of neuropeptide Y (NPY) in the reversal effect.

METHODSA rat model of anticonvulsant tolerance to FZP was prepared. The rats with FZP tolerance were randomly assigned to seven groups: FZP-tolerance, and nifedipine, levetiracetam, topiramate, flumazenil, L-NAME and pyridoxamine treatment groups. The tolerance to FZP was evaluated through pentylenetetrazol (PTZ) infusion into a tail vein. The latency to onset of clonic seizure and the PTZ threshold were recorded. The mRNA of NPY receptor Y2 in the hippocampus was determined by RT-PCR, and the distribution of NPY in the hippocampus was examined by immunohistochemistry.

RESULTSIn comparison with the blank control group, the average latency to the onset of clonic seizure was shortened, the average PTZ threshold decreased and the expression of NYT and NPY receptor Y2 mRNA decreased significantly in the FZP-tolerance group (p<0.01). In comparison with the FZP-tolerance group, the average latency to onset of clonic seizure was prolonged by 2 times and the average PTZ threshold doubled in the topiramate treatment group. The average latency to onset of clonic seizure was prolonged by 1 time and the average PTZ threshold increased 1 time in the nifedipine, the levetiracetam and the flumazenil treatment groups. The mRNA expression of NPY receptor Y2 increased by 1 or 2 times in the flumazenil, the nifedipine and the topiramate treatment groups when compared with the FZP-tolerance group.

CONCLUSIONSNifedipine, levetiracetam, topiramate and flumazenil can reverse the anticonvulsant tolerance to flurazepam. NPY may play a role in mediating the reversal effect.

Animals ; Anticonvulsants ; pharmacology ; Drug Tolerance ; Flurazepam ; pharmacology ; Hippocampus ; chemistry ; drug effects ; Male ; Neuropeptide Y ; analysis ; physiology ; Pentylenetetrazole ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Reaction Time ; Receptors, Neuropeptide Y ; genetics ; Seizures ; chemically induced ; drug therapy

OBJECTIVETo investigate the effects of alpha-keto acid on the expression of neuropeptide Y in malnutrition rats with chronic renal failure.

METHODSSD rats received 5/6 nephrectomy and were fed with 4% casein to establish models of malnutrition with chronic renal failure. Serum albumin, urea nitrogen, serum creatinine, type-1 insulin like growth factor and body weight of the rats were measured. The rat models were randomized into chronic renal failure group, alpha-keto acid group and normal control group, and after a 4-week treatment as indicated, neuropeptide Y mRNA levels in the hypothalamus were measured by RT-PCR in rats with surgically induced renal failure (two-stage subtotal nephrectomy). The blood neuropeptide Y of the rats were analyzed by radioimmunoassay.

RESULTSMalnutrition occurred in chronic renal failure rats at the end of 10 weeks. Compared with those in the chronic renal failure group, the plasma neuropeptide Y concentrations in alpha-keto acid group were significantly lowered with substantially elevated neuropeptide Y mRNA expression in the hypothalamus.

CONCLUSIONalpha-keto acid capsule can improve malnutrition in rats with renal insufficiency possibly by up-regulating neuropeptide Y mRNA expression in the hypothalamus and reducing the level of blood neuropeptide Y.

Animals ; Hypothalamus ; metabolism ; Keto Acids ; pharmacology ; therapeutic use ; Kidney Failure, Chronic ; blood ; Male ; Malnutrition ; blood ; drug therapy ; Neuropeptide Y ; blood ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo assay the expression of neuropeptide Y (NPY) in rat testes and to investigate the significance of NPY in the regulation of androgen production and spermatogenesis.

METHODSNPY mRNA levels in SD rat testes were measured by RT-PCR semi-quantity with beta-actin as internal control. NPY distribution was observed immunohistochemically.

RESULTSNPY gene expressed in the testes, showing the strong positive band in the PCR production electrophoresis gel. In the immunostaining slides, NPY was found positively expressed in the Leyding cell area and around the testicular vessels and tubules, but not in the seminiferous tubules.

CONCLUSIONThere was positive expression of NPY in the rat testes, which showed that NPY played a direct role in the regulation of testicular function.

Animals ; Immunohistochemistry ; Male ; Neuropeptide Y ; analysis ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Testis ; chemistry

The PTD-NPY fusion gene derived from HIV-1 TAT protein transduction domain and rat neuropeptide Y was amplified by overlap extension PCR, digested and subcloned into yeast expression vector pPICZ alpha A to construct recombinant expression plasmid pPICZ alpha-PTD-NPY. The cloned PTD-NPY fusion gene was identified by PCR and restriction enzyme digestion and sequenced. The exact recombinant plasmid was linearized by Sac I and integrated by electrotransformation into the genome of Pichia pastoris GS115 cells. Then, these positive recombinant yeast cells were induced by 10 mL/L methanol to express soluble PTD-NPY fusion protein. After 120 h of methanol induction, the SDS-PAGE electrophoresis result indicated PTD-NPY fusion protein was efficiently secreted into the medium. Western blotting analysis proved that the expressed fusion protein had specific NPY binding activity. The successful expression of PTD-NPY fusion protein in Pichia pastoris provided basis for its further application study.
Animals ; Cloning, Molecular ; Gene Fusion ; Genetic Vectors ; genetics ; Neuropeptide Y ; genetics ; Pichia ; genetics ; metabolism ; Polymerase Chain Reaction ; methods ; Rats ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transduction, Genetic ; tat Gene Products, Human Immunodeficiency Virus ; genetics

OBJECTIVETo investigate the effect of Shenshuai Yangzhen Capsule (SYC) on hypothalamic leptin-neuropeptide Y (NPY) and proopiomelanocortin (POMC) axes in chronic renal failure (CRF) rats with malnutrition (MN).

METHODSForty-two male SD rats of SPF grade were established into CRF-MN model by 5/6 nephrectomy and 4% casein diet, the happening time of MN in them was recorded. Rats successfully modeled were randomized into three groups, 11 rats in Group A treated with SYC, 11 in group B treated with composite alpha-keto acid and 12 in Group C was untreated. Besides, a normal control group was set up with 8 healthy rats. After being treated for 4 weeks, the renal function related indices, including serum creatinine (Scr), blood urea nitrogen (BUN), 24 hour urine protein (24 h Upro), albumin (ALB), haemoglobin (Hb) insulin like growth factor-1 (IGF-1), total cholesterol (TC) and triglyeride (TG) were measured, and body weight, food intake in rats were observed dynamically, blood leptin and NPY level in rats were determined by radioimmunoassay; mRNA expressions of OB-Rb, NPY and POMC in hypothalamus were detected with RT-PCR.

RESULTSCRF rats revealed MN at the end of 10th week after modeling. Compared with Group C, the condition of MN in Group A was significantly improved, showing increase of food intake and body weight (P < 0.05), marked improvement of renal function (P < 0.05), decrease of LP and NPY levels in plasma (P < 0.05), as well as up-regulated NPY mRNA expression and down-regulated mRNA expressions of OB-Rb and POMC in hypothalamus (P < 0.01).

CONCLUSIONSYC can improve the malnutrition condition in rats with CRF, which is possibly by way of depressing OB-Rb and POMC mRNA expression and upgrading NPY mRNA expression in hypothalamus.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Hypothalamus ; metabolism ; Kidney Failure, Chronic ; complications ; metabolism ; physiopathology ; Leptin ; genetics ; metabolism ; Male ; Malnutrition ; etiology ; metabolism ; physiopathology ; Neuropeptide Y ; genetics ; metabolism ; Pro-Opiomelanocortin ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the underlying mechanism of electroacupuncture for anti-aging.

METHODSForty Sprague-Dawley rats (female and male take one half for each), 3-month old, were divided into a control group, a model group, a routine electroacupuncture group (current intensity, 1 mA) and a strong electroacupuncture group (current intensity, 4.5 mA), 10 rats in each group. The model of aged rats was established by D-galactose in the latter three groups. The acupoints of "Guanyuan" (CV 4) and "Zusanli" (ST 36) were used for electroacupuncture treatment, six times per week for 4 weeks. After that, the level of interleukin 6 (IL-6) in the serum, as well as the expression of neuropeptide Y mRNA (NPY mRNA) and IL-6 receptor (IL-6R) in the periventricular hypothalamic nucleus (PVN) were examined and compared between each group.

RESULTSIn comparison of the control rats, the model rats expressed with the lower level of NPY mRNA in PVN, higher levels of IL-6 in the serum and IL-6R in PVN, which is different from each other (P < 0.05). In both routine electroacupuncture group and strong electroacupuncture group, the level of NPY mRNA in PVN was up-regulated, in contrast, the levels of IL-6 in the serum and IL-6R in PVN were cut down, which were different from those of the model group (both P < 0.05). Furthermore, the therapeutic effect of the strong electroacupuncture group is different from that of the routine electroacupuncture group (P < 0.05).

CONCLUSIONElectroacupuncture at "Qiangzhuang" acupoints plays an active role to slow down the aging process on the sub-acute aged rats through regulating the function of neuro-immune system, and the therapeutic effect of strong stimulation is better than that of routine stimulation.

Acupuncture Points ; Aging ; immunology ; physiology ; Animals ; Disease Models, Animal ; Electroacupuncture ; Female ; Humans ; Hypothalamus ; metabolism ; Interleukin-6 ; genetics ; metabolism ; Male ; Neuroimmunomodulation ; Neuropeptide Y ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Interleukin-6 ; genetics ; metabolism

OBJECTIVE: To screen the differentially expressed gene profile from the smooth muscles in the fundus uterus at the active stage of labor, and to provide candidate genes for picking out the drug targets related to uterine contraction. METHODS: Differentially expressed genes of uterine smooth muscles in the corpus from pro and post spontaneous parturition and those induced by oxytocin,as well as those from the corpus and the lower portion spontaneous parturition,were scanned respectively by human full-length genetic cDNA microarray with 8064 probe sets. Semi-quantitative RT-PCR was applied to testify the expression of voltage dependent calcium channel-L subtype (CACNA). The differentially expressed genes in the structure and function of the drug targets were picked out by bio-informatics to serve as candidate drug targets related to uterine contraction. RESULTS: The expressions of 29 genes were upregulated in fundus smooth muscles from the pro and post natural parturition, the pro and post inductive parturition of oxytocin, and the natural parturition. The expression of CACNA gene in RT-PCR was in accordance with that in the microarray. Among the 29 genes, neuromedin B receptor (NMBR) gene and neuropeptide Y (NPY) gene were the genes which not only had the targets of uterine contracted medicine, but also could contract the uterine. The differential expression ratios of NMBR in the above 3 types of uterine myometrium were 6.9,11.3, and 9.0, respectively while those of NPY were 6.0,29.8, and 2.9 respectively. CONCLUSION NMBR, whose expression in the uterine smooth muscles is always up-regulated at different parturition conditions, is likely to be an ideal candidate target of uterotonic drugs.
Calcium Channels ; genetics ; Drug Evaluation, Preclinical ; Female ; Gene Expression ; Gene Expression Profiling ; Humans ; Myometrium ; drug effects ; Neuropeptide Y ; genetics ; Oligonucleotide Array Sequence Analysis ; Pregnancy ; Receptors, Bombesin ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Uterine Contraction ; drug effects

OBJECTIVETo investigate the role of Ca(2+)/calmodulin-dependent calcineurin (CaN) signaling pathway in neuropeptide Y (NPY)-induced cardiomyocyte hypertrophy in rat.

METHODSCardiomyocytes of neonatal Wistar rats were cultured in the presence of 10 and 100 nmol/L NPY, and cyclosporine A (CsA) was applied to inhibit the activity of CaN. The protein synthesis rate, c-jun mRNA expression, CaN protein expression, CaN activity and intracellular Ca(2+) concentration in the cardiomyocytes were assessed.

RESULTSCompared with the control group, (3)H-Leu incorporation and expression of c-jun mRNA in the cardiomyocytes treated with 100 nmol/L NPY increased significantly (P<0.05, P<0.001), and the effect of NPY was blocked by CsA. The activity of CaN (P<0.05), CaN expression (P<0.05), and Ca(2+) concentration in the cytoplasm (P<0.001) and nuclei (P<0.001) of the cells with 100 nmol/L NPY treatment also significantly increased compared with those in the control cells.

CONCLUSIONNPY can induce cardiomyocyte hypertrophy in rats, in which process Ca(2+)/calmodulin-dependent CaN signaling pathway plays an important role.

Animals ; Animals, Newborn ; Calcineurin ; metabolism ; Cells, Cultured ; Hypertrophy ; Myocytes, Cardiac ; metabolism ; pathology ; Neuropeptide Y ; pharmacology ; Proto-Oncogene Proteins c-jun ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Signal Transduction