Brain ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression ; Neuropeptides ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-fos ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-jun ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics

OBJECTIVE: To explore the relationship between neuropeptides and mast cell in the initiation and development of allergic rhinitis. METHOD: Thirty healthy rats were randomly divided into three groups. The rat model of allergic rhinitis was established by using ovalbumin intraperitoneal immunization and nasal antigen challenge. After treating with capsaicin for two weeks, the counts of mast cells and the density of SP distribution were observed routinely in the nasal mucosa obtained from each models by HE, toluidine blue and immunohistochemical staining. RESULT: The counts of mast cells in AR were greatly more than them in normal controls (P < 0.01). After treating with capsaicin the mast cells were rare and significantly fewer than the normals (P < 0.01); The expression of SP was lower than the AR (P < 0.01), but no difference between the capsaicin group and normal group (P > 0.05). CONCLUSION Capsaicin can decrease the infiltration of mast cells, down regulate the SP expression, and improve the symptoms of AR greatly.
Animals ; Capsaicin ; therapeutic use ; Mast Cells ; metabolism ; Nasal Mucosa ; pathology ; Neuropeptides ; biosynthesis ; Rats ; Rats, Sprague-Dawley ; Rhinitis, Allergic, Perennial ; drug therapy ; metabolism ; pathology

OBJECTIVETo study the effect of ligustrazine on the migration of neuronal precursors (NPs) after focal cerebral ischemia in adult rats and explore its acting mechanism on recovery of function.

METHODSRat model of left middle cerebral artery occlusion (MCAO) was established by thread ligation. Ligustrazine 40 mg/kg was injected peritoneally once a day 2 h after modeling. On the 3rd, 7th, 14th and 21st day after operation, the migration of Doublecortin (DCX, the marker of NPs) in subventricular zone (SVZ) and the rostral migratory stream (RMS) were observed with immunohistochemistry.

RESULTSThe migration of DCX-positive cells in SVZ (abbrev. as migration below) through RMS into the olfactory bulb started from the 3rd day after ischemia, and lasted to the 21st day; the migration directly or through RMS into the ischemic penumbra of the adjacent striatum started on the 7th day, and increased significantly on the 14th day; and a few of DCX positive cells migrated through corpus callosum into the ischemic cortex on the 21st day. The migration was similar in the two groups in its pathway, but the extent in the ligustrazine group was more intensive.

CONCLUSIONLigustrazine could promote direct migration of NPs into the ischemic cerebral cortex and striatum, suggesting that it might play an important role in promoting self-recovery of brain function after ischemia through accelerating the migration of NPs.

Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Brain Ischemia ; physiopathology ; Cell Movement ; drug effects ; Immunohistochemistry ; Male ; Microtubule-Associated Proteins ; biosynthesis ; Neurons ; drug effects ; metabolism ; pathology ; Neuropeptides ; biosynthesis ; Pyrazines ; pharmacology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo evaluate the effects of acute glucose level changes on expression of prepro-orexin, orexin 1 receptor (OX1R) and orexin 2 receptor (OX2R) mRNA in rat hypothalamus tissue and pancreatic islets cells.

METHODSThirty adult male Wistar rats were randomly divided into three equal groups (n = 10). The acute hypoglycemia rat model was induced by a single subcutaneous injection of insulin. Twenty acute hypoglycemia rats were divided into group B and group C. Group B was allowed to eat freely, while group C was food-deprived. Control rats were injected the same volume of saline. The effect of glucose levels (2.8 mmol/L and 8.3 mmol/L) on pancreatic islet cell orexin system was detected in pancreas islet cell cultured in vitro. The expression of prepro-orexin and OXR mRNA was examined in rat hypothalamus tissue and pancreatic islets cell cultured in vitro using reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSExpression of orexin mRNA increased about 150% for the food-deprived hypoglycemia rats in comparison with control group (P < 0.01), whereas expression of OX1R mRNA decreased up to 30% (P < 0.01). However, expression of OX2R mRNA was unchanged in comparison with control group. In vitro, after incubation with 2.8 mmol/L glucose for 6 hours, the expression of prepro-orexin mRNA increased 2 times in rat pancreas islet cells in comparison with 8.3 mmol/L glucose group (P < 0.01). But the expression of OX1R mRNA was not sensitive to acute glucose fluctuation.

CONCLUSIONSOrexin in rat hypothalamus is stimulated by decline in blood glucose and inhibited by signals related to feeding. Moreover, glucose plays a role in modulating the gene expression of prepro-orexin in rat pancreatic islet cells.

Animals ; Blood Glucose ; metabolism ; Glucose ; pharmacology ; Hypoglycemia ; metabolism ; Hypothalamus ; metabolism ; Insulin ; pharmacology ; Intracellular Signaling Peptides and Proteins ; genetics ; Islets of Langerhans ; metabolism ; Male ; Neuropeptides ; biosynthesis ; genetics ; Orexin Receptors ; Orexins ; Protein Precursors ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Wistar ; Receptors, G-Protein-Coupled ; Receptors, Neuropeptide ; biosynthesis ; genetics

Pituitary adenylate cyclase activating polypeptide (PACAP) was first isolated from ovine hypothalamus and was known to stimulate the release of growth factor in various cells. Recently, we reported the cellular localization of PACAP and its type I (PAC1 ) receptor in rat placenta during pregnancy. Placenta is a critical organ that synthesizes several growth factors and angiogenic factors for the fetal development and its own growth. However, there is little information regarding the cellular localization of PACAP and its receptor in human placenta at various gestations. The aim of the present study was to define the expression and distribution of PACAP and PAC1 receptor mRNAs in the human placenta during the pregnancy period. PACAP and PAC1 receptor mRNAs were expressed in stroma cells of stem villi and terminal villi. At the early stage, on 7 and 14 weeks, PACAP and PAC1 receptor genes were moderately expressed in stroma cells surrounding the blood vessels within stem villi. These genes were strongly expressed in stroma cells of stem villi and terminal villi on 24 and 38 weeks. The expression of these genes was increased as gestation advanced, and localized in the same areas. Localization of PACAP and PAC1 receptor demonstrate the evidence that PACAP may play an important role, as an autoregulator or pararegulator via its PAC1 receptor. In conclusion, our findings strongly suggest that PACAP may have a critical role in physiological function of the placenta for gestational maintenance and fetal growth.
Chorionic Villi/metabolism ; Female ; Gene Expression ; Humans ; Nerve Growth Factors/*biosynthesis ; Neuropeptides/*biosynthesis ; Neurotransmitter Agents/*biosynthesis ; Pituitary Adenylate Cyclase-Activating Polypeptide ; Placenta/*metabolism ; Pregnancy ; Pregnancy Trimester, First ; Pregnancy Trimester, Second ; RNA, Messenger ; Receptors, Cell Surface/*biosynthesis ; Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide ; Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Type I

OBJECTIVETo construct recombinant lentiviral vectors carrying Rheb gene and its mutant Rheb'D60K gene, and examine their expression in human liver cancer cells.

METHODSRheb gene was amplified by PCR to construct the recombinant plasmid LV31-Rheb-WT and LV31-Rheb-D60K. HEK-293 FT cells were contransfected with the recombinant lentiviral vector together with a lentiviral package plasmid to produce the lentiviral particles. The expression of PS6 protein was detected in the lentivirus-infected MCF-7 cells. The apoptosis of SK-HEP-1 cells transfected with LV31-Rheb-WT or LV31-Rheb-D60K was observed.

RESULTSThe recombinant LV31-Rheb-WT and LV31-Rheb-D60K vectors were confirmed by PCR and DNA sequencing. Western blotting showed that PS6 protein expression was increased in LV31-Rheb-WT-transfected cells while decreased in LV31-Rheb-D60K-transfected cells. LV31-Rheb-D60K-transfected SK-HEP-1 cells showed more obvious apoptosis after starvation than LV31-Rheb-WT-transfected cells.

CONCLUSIONLentiviral vectors carrying Rheb gene and its mutant has been successfully constructed, which can be useful in further investigation of the role of Rheb gene in cancer cells.

Apoptosis ; genetics ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Genetic Vectors ; genetics ; HEK293 Cells ; Humans ; Lentivirus ; genetics ; metabolism ; Liver Neoplasms ; metabolism ; pathology ; MCF-7 Cells ; Monomeric GTP-Binding Proteins ; biosynthesis ; genetics ; Mutant Proteins ; genetics ; Neuropeptides ; biosynthesis ; genetics ; Ras Homolog Enriched in Brain Protein ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection

OBJECTIVETo investigate the effect of Shenmai Injection (SMI) and aminophylline on small airway smooth muscle cell (SASMC) apoptosis and the Fas/FasL expression in the papain induced emphysema model rats.

METHODSEmphysema model in rat was established by a single intratracheal instillation of papain. Apoptosis and Fas/FasL expression of SASMC were examined by immunohistochemical SABC and TUNEL assay at 1, 3, 5, 7, 15 and 30 days after modelling, and the effect of SMI and aminophylline on them were observed.

RESULTSFas, FasL expressions in normal SASMC were very low with a positive rate of (2.31 +/- 0.05)% and (1.28 +/- 0.47)% respectively. After papain instillation, the positive rates increased along with the prolonging of instillation time. SMI showed an inhibition on SASMC Fas and FasL expression but aminophylline didn't show. SASMC apoptosis was very low in normal rats with a rate of (0.87 +/- 0.32)%, it also raised after papain instillation and increased progressively along with the instillation time. SMI treatment could lower the apoptosis rate but aminophylline couldn't.

CONCLUSIONFas and FasL participated the SASMC apoptosis modulation in emphysema formation. SMI shows a definite treatment effect on emphysema by influencing the Fas and FasL protein expression and reducing SASMC apoptosis through inhibiting the release of inflammatory mediator.

Aminophylline ; pharmacology ; Animals ; Apoptosis ; drug effects ; Bronchi ; metabolism ; pathology ; Cells, Cultured ; Drug Combinations ; Drugs, Chinese Herbal ; pharmacology ; Emphysema ; metabolism ; pathology ; Fas Ligand Protein ; Female ; Male ; Membrane Glycoproteins ; biosynthesis ; metabolism ; Muscle, Smooth ; cytology ; metabolism ; Neuropeptides ; biosynthesis ; metabolism ; Papain ; Random Allocation ; Rats ; Rats, Wistar ; Receptors, Tumor Necrosis Factor ; fas Receptor

OBJECTIVETo explore the effect of intestinal ischemia/reperfusion (I/R) injury on leptin and orexin-A levels in peripheral blood and central secretory tissues, and investigate the roles of leptin and orexin-A in acute inflammatory responses.

METHODSAn intestinal I/R injury rat model was established, and the rats were grouped according to duration of the reperfusion time following a 60-min ischemia. Radioimmunoassay was used to examine the protein levels of leptin in the serum and adipose tissue, and the protein levels of orexin-A in the plasma and hypothalamus. Reverse transcriptase-polymerase chain reaction was also performed to detect the mRNA expressions of adipose leptin and hypothalamus orexin-A.

RESULTSCompared with that before injury, serum leptin level of 60-min ischemia with 30-min reperfusion (I60'R30') group decreased significantly and that of I60'R360' increased significantly. Compared with the sham-operation group (sham) after injury, serum leptin level of I60'R360' group increased significantly, and adipose leptin protein levels of I60'R30' and I60'R90' groups decreased significantly, whereas that of I60'R360' group increased obviously. Compared with sham group after injury, adipose leptin mRNA expressions of I60'R30', I60'R240' and I60'R360' groups all increased significantly, while that of I60'R150' showed significant decrease. No significant changes were noted in the protein levels of orexin-A either in the plasma or hypothalamus after I/R injury. In comparison with sham group after injury, hypothalamus orexin-A mRNA expressions of I60'R30' and I60'R90' groups showed gradual but significant decrease, and till 150 min of reperfusion, the expression reached its lowest, followed then by slow recovery at 240 and 360 min, though still remaining significantly lower than that of sham group.

CONCLUSIONLeptin and orexin-A have a time-dependent response to intestinal I/R injury, but the former appears to exhibit a faster response, and they may play a certain role in the metabolic disorders of acute inflammation.

Animals ; Female ; Inflammation ; blood ; genetics ; physiopathology ; Intestine, Small ; blood supply ; metabolism ; Intracellular Signaling Peptides and Proteins ; blood ; genetics ; Leptin ; blood ; genetics ; Male ; Neuropeptides ; blood ; genetics ; Orexins ; RNA, Messenger ; biosynthesis ; genetics ; Rabbits ; Radioimmunoassay ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; blood ; genetics ; physiopathology ; Reverse Transcriptase Polymerase Chain Reaction