untitled

Size: px

Start display at page:

Download "untitled"

庭炜任
5 years ago
Views:

1 Acta Physiologica Sinica, December 25, 2005, 57 (6): * (somatostatin, SST) 6 2 d 45 d SST SST SST 2 (somatostatin receptor 2, SSTR2) (1) 6 SST ( ) ng/mg SST SST SST ( ) ng/mg 6 (P<0.01) (2) SST (3) 6 SSTR2 SSTR2 SSTR2 SSTR2 (4) SST SSTR2 SST SST SST SSTR2 SST SST Q483; Q485; R574 Changes of somatostatin and expression of somatostatin receptor in small intestine and liver tissues during macaque development GUO Mei-Mei, TAN Qing-Hua, FAN Hua, HUANG Ming-Hui, WANG Chun-Hui, QIU Xiao-Qing, TANG Cheng-Wei * Division of Peptides Related with Human Diseases, State Key Laboratory of Biotherapy of Human Diseases, West China Hospital, Sichuan University, Chengdu , China Abstract: Intestinal tract, which produces more than fifty kinds of gut peptides, is regarded as the largest endocrine organ. With regard to the gut peptides, a number of studies were focused on their structure, function and the roles in some diseases. The changes in output or distribution of gut peptides in the intestinal tract during development have been largely unknown. This study was aimed to investigate the changes of somatostatin (SST) and somatostatin receptor 2 (SSTR2) in small intestinal and hepatic tissues during the development of macaque. The tissue samples of small intestine, liver or blood samples from peripheral and portal vein of 4 macaques in 6-month fetus, 2-day neonate, 45-day neonate and adult were obtained after anesthetization. The concentrations of SST in blood or tissues of macaques were measured by radioimmunoassay. The distributions of SST in small intestinal or hepatic tissues were visualized by immunohistochemical staining. The expression of SSTR2 was detected by in situ hybridization. SST concentration of intestinal tissue in 6-month-old macaque was (27.3±16.6) ng /mg protein and light positive staining of SST was localized in mucosal crypts but negative in muscle layer. The intestinal concentration of SST increased gradually with macaque development and reached to the peak [(120.1±35.3) ng /mg protein] in adult. It was significantly higher than that in fetus (P<0.01). Strong positive staining of SST was found in both mucosal crypts and myenteric nerve plexus of adult animal. SSTR2 was obviously expressed in intestinal epithelium of fetus but its expression was greatly reduced in epithelium and was shifted to mucosal crypts when grown to adult. Negative staining of Received Accepted This work was supported by the National Natural Science Foundation of China (No ). * Corresponding author. Tel: ; Fax: ; cwtang@medmail.com.cn

2 720 Acta Physiologica Sinica, December 25, 2005, 57(6): SSTR2 in muscle layer of fetal or neonatal macaque turned to be positive in myenteric nerve plexus of adult. The levels of SST or SSTR2 in liver decreased gradually during development. SST concentrations of small intestinal tissue kept significantly higher than those of hepatic tissues in the macaque developing stages. SST levels of portal vein were also maintained significantly higher than those of peripheral blood in the macaque developing stages. In conclusion, the level of SST and expression of SSTR2 in mucosal crypt increased gradually with macaque development. SST from intestinal tract was quickly degraded in portal vein before entering into liver. SST positive myenteric nerve plexus was visualized only in mature macaque. Key words: somatostatin; somatostatin receptor; development; macaque; small intestine; liver Brazeau [1] (somatostatin, SST) SST D [2] SST [3-6] SST [7-11] SST SST SST d 45 d (5 ) 1.2 ( ) 20~35 µl/kg 5 ml 3 ml ( 2~3 cm) ( ) ( ) 10% EDTA-Na 2 60 µl kiu r/min 15 min 0.5 ml/ 1.0 ml/ ml 50 µl 4 2 ml ml - (3:1 V/V) 3 ml mg 2 ml min 0.1 mol/l 0.5 ml (VC750 Bio- Rad ) 4 1~2 h 0.1 mol/l NaOH 0.5 ml r/min 30 min ml 3 ml (20~ 25 ) 15 min (550 Bio-Rad ) A 595nm 1.6 (radioimmunoassay, RIA) [12] SST ( ) (SN-695B ) SST 60 2 h 15 min / (37 ) 20 min 10% 20 min SST (Zymed ) 1: min 4 - (Zymed )

3 : 721 DAB (Santa Cruz ) 1.8 SST 2 (somatostain recepcer 2, SSTR2) 20% 3% DEPC 1% 37 4 h 37 SSC ( ) DAB 1.9 mean SD SST pg/mg SPSS10.0 P< SST SST SST SST 10~1 000 P<0.01 ( 1) SST P<0.01 SST ( 1) 2.2 SST SST SST SST SST SST ( 1) SST 45 d SST SST ( 2) SST 2.3 SSTR2 6 SSTR2 SSTR2 SSTR2 SSTR2 SSTR2 ( 3) 6 SSTR2 SSTR2 SSTR2 ( 4) 3 SST SST SST [12-14] SST SST SST SST SST Age 1. SST Table 1. SST concentration changes in blood or tissues during development of macaque Blood (pg/ml) Tissue (pg/mg protein) Peripheral blood Portal vein Liver Small intestine 6-month fetus ± ±9.54 * 2050±779 # 27272±16689 # 2-day neonate 0.85± ±1.74 * 1206± ± day neonate 46.70± ±3.72 * 404± ±4553 Adult ± ±71.08 * 115± ±35263 * P<0.01 compared to peripheral blood, P<0.01 compared to liver, # P<0.01 compared to adult.

4 722 Acta Physiologica Sinica, December 25, 2005, 57(6): SST ( ) Fig.1. Comparison of SST positive staining and distribution in small intestines of fetal and adult macaque (immunohistochemical staining). The arrows pointing to the brown staining show SST positive reaction. Scale bar, 1 mm. 2. SST ( ) Fig.2. Comparison of SST positive staining in liver during development of macaque (immunohistochemical staining). The arrows pointing to the brown staining show SST positive reaction. Scale bar, 0.1 mm. SST SST SST [15] SST SST [16,17] SST SST SSTR2 SSTR2 SST SSTR

5 : SSTR2 ( ) Fig.3. Comparison of SSTR2 positive staining and distribution in small intestines during development of macaque (in situ hybridization immunohistochemical staining). Scale bar, 1 mm. 4. SSTR2 ( ) Fig.4. Comparison of SSTR2 positive staining in liver during development of macaque (in situ hybridization immunohistochemical staining). Scale bar, 0.1 mm. D SST [18] SST SSTR2 Gomez de Segura [16] SST SST SSTR2 SST SSTR2 SST SSTR2 Abdu [19] SST SSTR2 SST SSTR2 SST

6 724 Acta Physiologica Sinica, December 25, 2005, 57(6): SST SST SST SST RIA SST SSTR2 SSTR2 [20] SSTR2 SSTR Brazeau P, Vale W, Burgus R, Ling N, Butcher M, Rivier J, Guillenim R. Hypothalamic polypeptide that inhibits the secretion of immunoreactive pituitary growth hormone. Science 1973; 179 (68): Ruan HZ ( ), Li XC, Cai WQ. Somatostatin inhibited pain modulation action of substance P in spinal cord. Acta Physiol Sin ( ) 1997; 49(1): (Chinese, English abstract). 3 Cooke HJ. Neurobiology of the intestinal mucosa. Gastroenterology 1986; 90(4): Bellinger DL, Lorton D, Romano TD, Olschowka JA, Felten SY, Felten DL. Neuropeptide innervation of lymphoid organs. Ann NY Acad Sci 1990; 594: Teitelbaum DH, Del Valle J, Reyas B, Post L, Gupta A, Mosely RL, Merion R. Intestinal intraepithelial lymphocytes influence the production of somatostation. Surgery 1996; 120: Ormsbee HS 3rd, Koehler SL Jr, Telford GL. Somatostatin inhibits motilin-induced interdigestive contractile activity in the dog. Am J Dig Dis 1978; 23(9): Kusumoto Y, Iwanga T, Ito S, Fujita T. Juxtaposition of somatostatin cell in the dog stomach. Arch Histol Jpn 1979; 42(4): Park J, Chiba T, Yamada T. Mechanisms for muscarinic for direct inhibition of canine gastric parietal cells by somatostatin. J Biol Chem 1987; 262(29): Seal A, Yamada T, Debas H, Hollinshead J, Osadchey B, Aponte G, Walsh J. Somatostatin-14 and -28: clearance and potency on gastric function in dogs. Am J Physiol 1982; 243 (2): G97-G Colturi TJ, Unger RH, Feldman M. Role of circulating somatostatin in regulation of gastric acid secretion, gastrin release, and islet cell function. Studies in healthy subjects and duodenal ulcer patients. J Clin Invest 1984; 74(2): Ferjoux G, Bousquet C, Cordelier P, Benali N, Lopez F, Rochaix P, Buscail L, Susini C. Signal transduction of somatostatin receptors negatively controlling cell proliferation. J Physiol Paris 2000; 94(3-4): Wang L ( ), Tang CW, Wang CH, Li X. Quantitative variation of vasoactive intestinal polypeptide and its receptor in rat liver during development. Acta Physiol Sin ( ) 2005; 57(3): (Chinese, English abstract). 13 Ebeid AM, Escourrou J, Soeters PB, Murray P, Fischer JE. Hepatic inactivation of vasoactive intestinal peptide in man and dog. Ann Surg 1978; 188(1): Svoboda M, De Neef P, Tastenoy M, Christophe J. Molecular characteristics and evidence for internalization of vasoactive intestinal peptide (VIP) receptors in the tumoral rat pancreatic cell line AR 4-2 J. Eur J Biochem 1988; 176(3): Lu B, Fifini M, Emanueli C, Geppetti P, Grady EF, Gerard NP, Ansell J, Payan DG, Gerard C, Bunnett N. The control of microvascular permeability and blood pressure by neutral endopeptidase. Nat Med 1997; 3(8): Gomez de Segura IA, Castell T, Vazquez I, Mata A, De Miguel E. Somatostatin antagonism prevents elemental diet-induced intestinal atrophy in the rat. Dig Dis Sci 2001; 46(9): Hajri A, Aprahamian M, Damage C. Effect of prolonged administration of long-acting somatostatin on caerulein, CCK-8 and GRP induced pancreatic growth in the rat. Regul Pep 1991; 32(2): Walters JR. Cell and molecular biology of the small intestine: new insights into differentiation, growth and repair. Curr Opin Gastroenterol 2004; 20(2): Abdu F, Hicks GA, Hennig G, Allen JP, Grundy D. Somatostatin sst(2) receptors inhibit peristalsis in the rat and mouse jejunum. Am J Physiol Gastrointest Liver Physiol 2002; 282 (4): G624-G Dasgupta P. Somatostatin analogues: multiple roles in cellular proliferation, neoplasia, and angiogenesis. Pharmacol Ther 2004; 102(1):

21 13 D / 2 10 ig ig 10 d 1. 0 g kg /2 2 β- 5 g kg g kg mg kg - 1 ig ig 24 h 5% 1 2% CMC-Na 2. 5 ml 30

21 13 D / 2 10 ig ig 10 d 1. 0 g kg /2 2 β- 5 g kg g kg mg kg - 1 ig ig 24 h 5% 1 2% CMC-Na 2. 5 ml 30 21 13 1 2 1* 2 1. 430065 2. 445000 20 10 5 g kg - 1 5 mg kg - 1 ig 10 d ig HE 1 TFF1 TFF1 P < 0. 01 P < 0. 01 P < 0. 01 P < 0. 05 R285. 5 doi 10. 13422 /j. cnki. syfjx. 2015130100 A 1005-9903 2015 13-0100-05