Effect of Ghrelin on Insulin Secretion and Expression of Glut-2 Protein in Isolated Pancreas of rat_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

YU Yifan , ZHOU Yong , WANG Yong ,  LIU Jingang

Department of Biliary， Hernia， and Weight Reduction Surgery， Affiliated Shengjing Hospital， China Medical University， Shenyang 110004， Liaoning Province， China;

Corresponding author：

LIU Jingang, Email: liujg51347@hotmail.com

Keywords：

Ghrelin; Glucose transporter protein-2; Isolated pancreas perfusion; Islet; Rat

Video：

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Objective 　To explore the effect of ghrelin on insulin secretion and expression of glucose transporter protein-2 （Glut-2） in isolated pancreas of rats.
Methods 　Twenty five Wistar rats were randomly devided into normal control group （NC group）， high concentration of glucose group （HCG group）， high concentration of glucose with high concentration of ghrelin group （10-8mol/L， HCG＋HCGh group）， medium concentration of ghrelin group（10-9mol/L， HCG＋MCGh group）， and low concentration of ghrelin group （10-10mol/L， HCG＋LCGh group） with 5 rats in each group. The rat isolated pancreas perfusion models were established firstly， then from the distal end of abdominal aortas， the models were perfused with low concentration of glucose （5.5mmol/L）， high concentration of glucose （33.3mmol/L） or high concentration of glucose added with different concentrations of ghrelin. Levels of insulin outflowed from portal vein were tested by ELISA method， expression levels of Glut-2 protein were tested by immunohistochemical method，and ultrastructure changes of islet β cell were observed under the transmission electron microscope.
Results 　There were no significant difference on levels of fasting blood glucose （FBG）， fasting insulins （FINS）， homeostasis model of assess-ment for insulin resistence index （HOMA-IR）， and homeostasis model of assessment for pancreatic β cell function （HOMA-β），（P＞0.05）. There were no significant difference on insulin levels of effluent from portal vein of 5 groups （P＞0.05） when isolated pancreas perfused with 5.5mmol/L glucose， while had 2 secretion peaks in 3min and 10-12min after 33.3 mmol/L glucose perfusion， where HCG＋HCGh group at the top. The mean density value of Glut-2 protein in NC group was higher than that of other 4 groups （P＜0.05）. The results of transmission electron microscopy showed that apoptosis was lighter in NC group than that of other 4 groups， and apoptosis of HCG＋HCGh group was lighter than that of HCG＋MCGh group and HCG＋LCGh group.
Conclusions 　In isolated pancreas of rats， ghrelin promotes high concentration of glucose-stimulated insulin secretion， decreases expression of Glut-2 protein， and protects the islet β cell.

Citation： YU Yifan,ZHOU Yong,WANG Yong,LIU Jingang,.. Effect of Ghrelin on Insulin Secretion and Expression of Glut-2 Protein in Isolated Pancreas of rat. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2013, 20(3): 251-258. doi: Copy

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7.	Pöykkö SM， Kellokoski E， Hörkkö S， et al. Low plasma ghrelin is associated with insulin resistance， hypertension， and the prevalence of type 2 diabetes[J]. Diabetes， 2003， 52(10)：2546-2553.
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12.	Date Y， Nakazato M， Hashiguchi S， et al. Ghrelin is present in pancreatic alpha-cells of humans and rats and stimulates insulin secretion[J]. Diabetes， 2002， 51(1)：124-129.
13.	Lee HM， Wang G， Englander EW， et al. Ghrelin， a new gastrointestinal endocrine peptide that stimulates insulin secretion：entericdistribution， ontogeny， influence of endocrine， and dietarymanipulations[J]. Endocrinology， 2002， 143(1)：185-190.
14.	Salehi A， Dornonville de la Cour C， Håkanson R， et al. Effects of ghrelin on insulin and glucagon secretion：a study of isolated pancreatic islets and intact mice[J]. Regul Pept， 2004， 118(3)：143-150.
15.	Chung H， Kim E， Lee DH， et al. Ghrelin inhibits apoptosis in hypothalamic neuronal cells during oxygen-glucose deprivation[J]. Endocrinology， 2007， 148(1)：148-159.
16.	Miao Y， Xia Q， Hou Z， et al. Ghrelin protects cortical neuron against focal ischemia/reperfusion in rats[J]. Biochem Biophys Res Commun， 2007， 359(3)：795-800.
17.	Thorens B， Wu YJ， Leahy JL， et al. The loss of GLUT2 expression by glucose-unresponsive beta cells of db/db mice is reversible and is induced by the diabetic environment[J]. J Clin Invest， 1992， 90(1)：77-85.
18.	Bell GI， Kayano T， Buse JB， et al. Molecular biology of mammalian glucose transporters[J]. Diabetes Care， 1990， 13(3)：198-208.
19.	Fukumoto H， Kayano T， Buse JB， et al. Cloning and characterization of the major insulin-responsive glucose transporter expressed in human skeletal muscle and other insulin-responsive tissues[J]. J Biol Chem， 1989， 264(14)：7776-7779.
20.	Thorens B， Cheng ZQ， Brown D， et al. Liver glucose transporter：a basolateral protein in hepatocytes and intestine and kidney cells[J]. Am J Physiol Cell Physiol， 1990， 259(6 Pt 1)：C279-C285.
21.	Thorens B， Sarkar HK， Kaback HR， et al. Cloning and functional expression in bacteria of a novel glucose transporter present in liver， intestine， kidney， and beta-pancreatic islet cells[J]. Cell， 1988， 55(2)：281-290.
22.	Gould GW， Thomas HM， Jess TJ， et al. Expression of human glucose transporters in Xenopus oocytes：kinetic characterization and substrate specificities of the erythrocyte， liver， and brain isoforms[J]. Biochemistry， 1991， 30(21)：5139-5145.
23.	Orci L， Thorens B， Ravazzola M， et al. Localization of the pancreatic beta cell glucose transporter to specific plasma membrane domains[J]. Science， 1989， 245(4915)：295-297.
24.	Guillam MT， Hümmler E， Schaerer E， et al. Early diabetes and abnormal postnatal pancreatic islet development in mice lacking Glut-2[J]. Nat Genet， 1997， 17(3)：327-330.
25.	Guillam MT， Dupraz P， Thorens B. Glucose uptake，utilization，and signaling in GLUT2-null islets[J]. Diabetes， 2000， 49(9)：1485-1491.
26.	Guillemain G， Muñoz-Alonso MJ， Cassany A， et al. Karyopherin alpha2：a control step of glucose-sensitive gene expression in hepatic cells[J]. Biochem J， 2002， 364(Pt 1)：201-209.
27.	， 86(10)：5083-5086.

1. Couzin J. Medicine. Bypassing medicine to treat diabetes[J]. Science， 2008， 320(5875)：438-400.
2. Henquin JC， Ravier MA， Nenquin M， et al. Hierarchy of the beta-cell signals controlling insulin secretion[J]. Eur J ClinInvest， 2003， 33(9)：742-750.
3. Wang Y， Liu J. Combination of bypassing stomach and vagus dissection in high-fat diet-induced obese rats—a long-term investigation[J]. Obes Surg， 2010， 20(3)：375-379.
4. Irako T， Akamizu T， Hosoda H， et al. Ghrelin prevents development of diabetes at adult age in streptozotocin-treated newborn rats[J]. Diabetologia， 2006， 49(6)：1264-1273.
5. Kojima M， Hosoda H， Date Y， et al. Ghrelin is a growth-hormone-releasing acylated peptide from stomach[J]. Nature， 1999， 402(6762)：656-660.
6. Mclaughlin T， Abbasi F， Lamendola C， et al. Plasma ghrelin concentrations are decreased in insulin-resistant obese adults relativeto equally obese insulin-sensitive controls[J]. Clin Endocrinol Metab， 2004， 89(4)：1630-1635.
7. Pöykkö SM， Kellokoski E， Hörkkö S， et al. Low plasma ghrelin is associated with insulin resistance， hypertension， and the prevalence of type 2 diabetes[J]. Diabetes， 2003， 52(10)：2546-2553.
8. 赵乃倩，余叶蓉，谭惠文，等. 大鼠胰腺离体灌注技术的建立 [J]. 中华内分泌代谢杂志， 2007， 23(5)：459-463.
9. Broglio F， Arvat E， Benso A， et al. Ghrelin，a natural GH secretagogue produced by the stomach，induces hyperglycemia andreduces insulin secretion in humans[J]. J Clin Endocrinol Metab，.
10. Egido EM， Rodriguez-Gallardo J， Silvestre RA， et al. Inhibitory effect of ghrelin on insulin and pancreatic somatostatin secretion[J]. Eur J Endocrinol， 2002， 146(2)：241-244.
11. Reimer MK， Pacini G， Ahrén B. Dose-dependent inhibition by ghrelin of insulin secretion in the mouse[J]. Endocrinology， 2003， 144(3)：916-921.
12. Date Y， Nakazato M， Hashiguchi S， et al. Ghrelin is present in pancreatic alpha-cells of humans and rats and stimulates insulin secretion[J]. Diabetes， 2002， 51(1)：124-129.
13. Lee HM， Wang G， Englander EW， et al. Ghrelin， a new gastrointestinal endocrine peptide that stimulates insulin secretion：entericdistribution， ontogeny， influence of endocrine， and dietarymanipulations[J]. Endocrinology， 2002， 143(1)：185-190.
14. Salehi A， Dornonville de la Cour C， Håkanson R， et al. Effects of ghrelin on insulin and glucagon secretion：a study of isolated pancreatic islets and intact mice[J]. Regul Pept， 2004， 118(3)：143-150.
15. Chung H， Kim E， Lee DH， et al. Ghrelin inhibits apoptosis in hypothalamic neuronal cells during oxygen-glucose deprivation[J]. Endocrinology， 2007， 148(1)：148-159.
16. Miao Y， Xia Q， Hou Z， et al. Ghrelin protects cortical neuron against focal ischemia/reperfusion in rats[J]. Biochem Biophys Res Commun， 2007， 359(3)：795-800.
17. Thorens B， Wu YJ， Leahy JL， et al. The loss of GLUT2 expression by glucose-unresponsive beta cells of db/db mice is reversible and is induced by the diabetic environment[J]. J Clin Invest， 1992， 90(1)：77-85.
18. Bell GI， Kayano T， Buse JB， et al. Molecular biology of mammalian glucose transporters[J]. Diabetes Care， 1990， 13(3)：198-208.
19. Fukumoto H， Kayano T， Buse JB， et al. Cloning and characterization of the major insulin-responsive glucose transporter expressed in human skeletal muscle and other insulin-responsive tissues[J]. J Biol Chem， 1989， 264(14)：7776-7779.
20. Thorens B， Cheng ZQ， Brown D， et al. Liver glucose transporter：a basolateral protein in hepatocytes and intestine and kidney cells[J]. Am J Physiol Cell Physiol， 1990， 259(6 Pt 1)：C279-C285.
21. Thorens B， Sarkar HK， Kaback HR， et al. Cloning and functional expression in bacteria of a novel glucose transporter present in liver， intestine， kidney， and beta-pancreatic islet cells[J]. Cell， 1988， 55(2)：281-290.
22. Gould GW， Thomas HM， Jess TJ， et al. Expression of human glucose transporters in Xenopus oocytes：kinetic characterization and substrate specificities of the erythrocyte， liver， and brain isoforms[J]. Biochemistry， 1991， 30(21)：5139-5145.
23. Orci L， Thorens B， Ravazzola M， et al. Localization of the pancreatic beta cell glucose transporter to specific plasma membrane domains[J]. Science， 1989， 245(4915)：295-297.
24. Guillam MT， Hümmler E， Schaerer E， et al. Early diabetes and abnormal postnatal pancreatic islet development in mice lacking Glut-2[J]. Nat Genet， 1997， 17(3)：327-330.
25. Guillam MT， Dupraz P， Thorens B. Glucose uptake，utilization，and signaling in GLUT2-null islets[J]. Diabetes， 2000， 49(9)：1485-1491.
26. Guillemain G， Muñoz-Alonso MJ， Cassany A， et al. Karyopherin alpha2：a control step of glucose-sensitive gene expression in hepatic cells[J]. Biochem J， 2002， 364(Pt 1)：201-209.
27. ， 86(10)：5083-5086.

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CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Effect of Ghrelin on Insulin Secretion and Expression of Glut-2 Protein in Isolated Pancreas of rat

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