The present study was aimed to investigate the changes of vasoactive intestinal polypeptide (VIP) and VIP receptor 1 (VIPR1) in small intestinal and hepatic tissues during macaque development. The tissue samples of small intestine, liver and blood samples from peripheral and portal vein of 4 macaques of 6-month fetus, 2-day neonate, 45-day neonate and adult were obtained after anesthetization. The concentration of VIP in blood or tissues of macaques was measured by radioimmunoassay. The distribution of VIP in small intestinal or hepatic tissues was visualized by immunohistochemical staining. The expression of VIPR1 was detected by in situ hybridization. The results showed that: (1) VIP concentration in intestinal tissue of 6-month fetus was (20.7+/-14.3) ng/mg protein, and a few VIP-positive nerve fibers first appeared in intestinal villus root and submucosal layer but not in muscle layer. The intestinal concentration of VIP increased gradually with macaque development and reached (514.8+/- 49.2) ng/mg protein in adult, significantly higher than that in 6-month fetus (P<0.01). (2) In adult animal, VIP-positive nerve fibers became thicker and gradually extended into the mucosal crypt, submucosal layer nerve, myenteric nerve plexus of annular muscle and indulge muscle, and annular muscle. Correspondingly, the expression of VIPR1 in intestine was up-regulated during development. (3) On the contrary, the levels of VIP and VIPR1 in liver were gradually decreased during development. (4) VIP concentration in small intestinal tissue was higher than that in hepatic tissue during development. The VIP level in portal vein was also significantly higher than that in peripheral blood during development. In conclusion, the levels of VIP and VIPR1 in mucosal crypt, submucosal layer nerve, myenteric nerve plexus of annular muscle and indulge muscle increase rapidly after birth. Most of VIP from intestinal tract is degraded in portal vein before entering liver, suggesting that VIP does not metabolize and decompose in liver, and that VIPR1 is only present in embryo hepatic blood vessels.
Animals ; Animals, Newborn ; Fetus ; Intestine, Small ; metabolism ; Liver ; metabolism ; Macaca mulatta ; embryology ; growth & development ; metabolism ; Receptors, Vasoactive Intestinal Polypeptide, Type I ; metabolism ; Vasoactive Intestinal Peptide ; metabolism

It was hypothesized that the VPAC1 agonist may exert anti-obesity functions because VPAC1 is involved in the anorexigenic effects and the anti-inflammatory function of pituitary adenylate cyclase-activating polypeptide (PACAP)/vasoactive intestinal polypeptide (VIP). Furthermore, our in vitro test showed that the expression of VPAC1 increased significantly after the 3T3-L1 adipocytes were differentiated, and that incubation of adipocytes with VPAC1 agonist (10-1 000 nmol/L per 1x10(6) cells) resulted in stimulation of lipolysis. To test the effect of VPAC1 agonist [Lys15, Arg16, Leu27]-VIP (1-7) GRF (8-27) on diet-induced obesity (DIO), we further designed the following two in vivo experiments: (1) Mice were fed on high-fat diet (HFD) and intraperitoneally (i.p.) treated with VPAC1 agonist simultaneously for 28 d; (2) Mice were given HFD for 35 d, and subsequently fed on the same HFD and i.p. treated with VPAC1 agonist for the next 28 d. The physiological indices, including body weight, weight of white adipose tissue, plasma glucose and blood lipid, were collected. The results showed that treatment with VPAC1 agonist inhibited ingestion significantly and prevented the elevations in body weight and the weights of the white adipose tissues (epididymal and dorsal) induced by HFD. The increases in plasma glucose, cholesterol, triglycerides and LDL induced by HFD were also down-regulated in mice treated with VPAC1 agonist. VPAC1 agonist treatment also improved the glucose tolerance. Therefore, VPAC1 agonist treatment inhibits the development of the obesity induced by HFD and helps to improve the morbidities associated with DIO.
3T3-L1 Cells ; Adipocytes ; drug effects ; Animals ; Body Weight ; Diet, High-Fat ; Mice ; Obesity ; drug therapy ; Receptors, Vasoactive Intestinal Polypeptide, Type I ; agonists ; Vasoactive Intestinal Peptide ; pharmacology

OBJECTIVE: To determine whether chronic alcoholism alters the expression levels of Vasoactive intestinal polypeptide (VIP) and its receptor (VIPR1) in the cochlea of chronic alcoholism rats. METHOD: We measured their expression levels in 30 SD rats, in which we created models of different degrees of chronic alcoholism. We investigated the presence of the mRNA of VIP in the cochlea of chronic alcoholism rats and controls by reverse transcription-polymerase chain reaction (RT-PCR) method. We investigated the presence of proteins of VIPR1 in poisoned rats and controls by western blot. We also evaluated the local distribution of VIP cells by immunohistochemistry. RESULT: We found that the levels of VIP and VIPR1 were downregulated in the chronic alcoholism groups compared to the controls group. The differences in some expression levels were significant different between chronic alcoholism rats and control rats. Moreover, at different degrees of alcohol poisoning in rats, the contents of VIP and VIPR1 differed. Decreased levels of VIP and VIPR1 were detected in the deep chronic alcoholism group compared to the group with low-degree poisoning (P < 0.05). In spiral ganglion cell plasm the expression of VIP and VIPR1 had no significant difference in three groups (P > 0.05). CONCLUSION These results suggest that VIP and VIPR1 play an important role in the auditory function in rats with chronic alcoholism. Chronic alcoholism may cause a peptide hormone secretion imbalance in the auditory system, eventually leading to hearing loss.
Alcoholism ; metabolism ; Animals ; Cochlea ; metabolism ; Disease Models, Animal ; Down-Regulation ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Receptors, Vasoactive Intestinal Polypeptide, Type I ; metabolism ; Spiral Ganglion ; Vasoactive Intestinal Peptide ; metabolism

OBJECTIVE: To explore the transcriptional regulation mechanism and biological function of low expression of vasoactive intestinal peptide receptor 1 (VIPR1) in hepatocellular carcinoma (HCC). METHODS: We constructed plasmids carrying wild-type VIPR1 promoter or two mutant VIPR1 promoter sequences for transfection of the HCC cell lines Hep3B and Huh7, and examined the effect of AP-2<i>αi> expression on VIPR1 promoter activity using dual-luciferase reporter assay. Pyrosequencing was performed to detect the changes in VIPR1 promoter methylation level in HCC cells treated with a DNA methyltransferase inhibitor (DAC). Chromatin immunoprecipitation was used to evaluate the binding ability of AP-2<i>αi> to VIPR1 promoter. Western blotting was used to assess the effect of AP-2<i>αi> knockdown on VIPR1 expression and examine the differential expression of VIPR1 in the two cell lines. The effects of VIPR1 overexpression and knockdown on the proliferation, cell cycle and apoptosis of HCC cells were analyzed using CCK8 assay and flow cytometry. We also observed the growth of HCC xenograft with lentivirus-mediated over-expression of VIPR1 in nude mice. RESULTS: Compared with the wild-type VIPR1 promoter group, co-transfection with the vector carrying two promoter mutations and the AP-2<i>αi>-over-expressing plasmid obviously restored the luciferase activity in HCC cells (<i>Pi> < 0.05). DAC treatment of the cells significantly decreased the methylation level of VIPR1 promoter and inhibited the binding of AP-2<i>αi> to VIPR1 promoter (<i>Pi> < 0.01). The HCC cells with AP-2<i>αi> knockdown showed increased VIPR1 expression, which was lower in Huh7 cells than in Hep3B cells. VIPR1 overexpression in HCC cells caused significant cell cycle arrest in G2/M phase (<i>Pi> < 0.01), promoted cell apoptosis (<i>Pi> < 0.001), and inhibited cell proliferation (<i>Pi> < 0.001), while VIPR1 knockdown produced the opposite effects. In the tumor-bearing nude mice, VIPR1 overexpression in the HCC cells significantly suppressed the increase of tumor volume (<i>Pi> < 0.001) and weight (<i>Pi> < 0.05). CONCLUSION VIPR1 promoter methylation in HCC promotes the binding of AP-2<i>αi> and inhibits VIPR1 expression, while VIPR1 overexpression causes cell cycle arrest, promotes cell apoptosis, and inhibits cell proliferation and tumor growth.
Animals ; Carcinoma, Hepatocellular/pathology* ; Cell Line, Tumor ; Cell Proliferation/genetics* ; Gene Expression Regulation, Neoplastic ; Humans ; Liver Neoplasms/pathology* ; Luciferases/genetics* ; Methylation ; Mice ; Mice, Nude ; Receptors, Vasoactive Intestinal Polypeptide, Type I/metabolism* ; Transcription Factor AP-2/metabolism*