Ghrelin对乙醇诱导的急性胃黏膜损伤的保护作用及机理研究
摘要
目的
研究ghrelin对乙醇诱导的急性大鼠胃黏膜损伤的保护作用,并通过观察辣椒素神经拮抗剂对ghrelin保护作用的影响来进一步探讨ghrelin的作用机制。
方法
实验一
1.64只雌性SD大鼠分成四组,空白对照组(2m1/kg 0.9%氯化钠液腹腔注射+1ml/100g 0.9%氯化钠液灌胃),乙醇模型组(2m1/kg 0.9%氯化钠液腹腔注射+1ml/10Og 75%乙醇灌胃),低剂量ghrelin组(2p,g/kg ghrelin腹腔注射+lml/100g 75%7,醇灌胃),高剂量ghrelin组(20μg/kg ghrelin腹腔注射+lml/100g 75%乙醇灌胃)
2小时后四组大鼠麻醉取胃。
2.计算各组大鼠胃黏膜溃疡指数。
3.HE染色观察胃黏膜炎症损伤程度。
4.通过荧光半定量RT-PCR法测定eNOS/iNOS和COX-1/COX-2的mRNA表达,
实验二
1.64只雌性SD大鼠分成四组,空白对照组(2m1/kg 0.9%氯化钠液腹腔注射+lml/100g 0.9%氯化钠液灌胃),乙醇模型组(2ml/kg 0.9%氯化钠液腹腔注射+1ml/100g 75%乙醇灌胃),ghrelin组(20μg/kg ghrelin腹腔注射+lml/100g 75%7,醇灌胃),Capsazepine组(20μg/kg ghrelin腹腔注射+1mg/kg Capsazepine腹腔注射+1ml/100g 75%乙醇灌胃),2小时后四组大鼠麻醉取胃。
2.计算各组大鼠胃黏膜溃疡指数。
3.HE染色观察胃黏膜炎症损伤程度。
4.通过荧光半定量RT-PCR法测定eNOS(?)(?)iNOS(?)(?)COX-1/COX-2的1nRNA表达,测定CGRP的mRNA的表达。
5.酶联免疫吸附试验(ELISA)法测定胃组织中CGRP含量。
结果
实验一
1. Ghrelin组胃溃疡指数明显小于乙醇模型组,与Ghrelin呈剂量相关性。
2. Ghrelin组病理炎症反应较乙醇模型组明显减轻,与Ghrelin呈剂量相关性。
3.荧光半定量RT-PCR:ghrelin组与乙醇模型组相比,eNOS mRNA表达上升,COX-1 mRNA表达无变化,但iNOS mRNA和COX-2表达下降,与ghrelin呈剂量相关性。
实验二
1. Capsazepine组胃溃疡指数明显大于ghrelin组。
2. Capsazepine组病理炎症反应明显较ghrelin组加重。
3.荧光半定量RT-PCR:capsazepine组与ghrelin组相比,iNOS和COX-2mRNA表达上升；(;NOS(?)(?)CGRP mRNA表达下降,COX-1 mRNA表达无变化
4.酶联免疫吸附试验(ELISA) CGRP测定：capsazepine组与ghrelin组相比,组织中CGRP含量下降。
结论
本实验结果提示Ghrelin具有对乙醇诱导的胃黏膜保护作用,其保护机制可能是通过参与辣椒素一敏感感觉神经保护性反射,调节CGRP的表达从而参与调控NOS和COX活性来达到胃黏膜保护的作用。
Objective
To investigate the gastroprotective effects of ghrelin against ethanol-induced acute gastric injury in rats and possible mechanism.
Methods
Part 1
1.64 female Sprague-Dawley(SD) rats were divided into four groups and were intragastrically administrated as follows:Control group(2mg/kg 0.9%NaCl ip+lml/100g 0.9%NaCl ig);Ethanol model group(2ml/kg 0.9%NaCl ip+lml/100g 75%ethanol ig);Low-dose ghrelin group(2μg/kg ghrelin ip+lml/100g 75%ethanol ig);High-dose ghrelin group(20μg/kg ghrelin ip+lml/100g 75%ethanol ig).Rats were anaesthetized and the stomachs were removed after 2 hours.
2. The ulcer index of gastric mucosal lesions in rats were calculated.
3. Gastric mucosal inflammation were assessed by hematoxylin-eosin (H&E) staining.
4. eNOS/iNOS and COX-1/COX-2 mRNA expression in these specimens were verified by semi-quantitative reverse transcriptional polymerase chain reaction (SqRT-PCR).
Part 2
1.64 female SD rats were divided into four groups and were intragastrically administrated as follows:Control group(2mg/kg 0.9%NaCl ip+lml/100g 0.9%NaCl ig);Ethanol model group(2ml/kg 0.9%NaCl ip+lml/100g 75%ethanol ig);Ghrelin group(2μg/kg ghrelin ip+lml/100g 75%ethanol ig);Capsazepine group(20μg/kg ghrelin ip+lmg/kg Capsazepine ip+lml/100g 75%ethanol ig).Rats were anaesthetized and the stomachs were removed after 2 hours.
2. The ulcer index of gastric mucosal lesions in rats were calculated.
3. The inflammatory lesions of gastric mucosa were assessed by hematoxylin-eosin (H&E) staining.
4. eNOS/iNOS, COX-1/COX-2 and CGRP mRNA expression in these specimens were verified by SqRT-PCR
5.The levels of CGRP in these specimens were measured by Enzyme-linked immunosorbent assay (ELISA).
Results
Part 1
1. The ulcer index of ghrelin groups were significantly less than that of ethanol model group, and were positively correlated with dosages of ghrelin.
2. Compared with ethanol model group,the gastric mucosal inflammation lesions of ghrelin group were significantly reduced, and were positively correlated with dosages of ghrelin.
3. Compared with Ethanol model group,grhrein groups were characterized by no changes of COX-1 mRNA level, but iNOS mRNA and COX-2 mRNA decreased, eNOS mRNA increased in a dose-dependent manner.
Part 2
1. The ulcerative index of capsazepine group were significantly greater than those of ghrelin group.
2. Compared with ghrelin group,the gastric mucosal inflammation lesions of capsazepine group increased significantly.
3. Compared with ghrelin group, there was no change of COX-1 mRNA level in capsazepine group,but mRNA levels of COX-2 and iNOS increased while eNOS and CGRP decreased.
4. Compared with ghrelin group, the levels of CGRP in gastric tissue of capsazepine decreased.
Conclusions
Ghrelin had protective effects on ethanol-induced acute gastric mucosal injury in rats.The mechanism of the protective effects was related to CGRP which can be involved in regulating capsaicin-sensitive nerves and thus participated in the regulation of the activity of NOS and COX.
研究ghrelin对乙醇诱导的急性大鼠胃黏膜损伤的保护作用,并通过观察辣椒素神经拮抗剂对ghrelin保护作用的影响来进一步探讨ghrelin的作用机制。
方法
实验一
1.64只雌性SD大鼠分成四组,空白对照组(2m1/kg 0.9%氯化钠液腹腔注射+1ml/100g 0.9%氯化钠液灌胃),乙醇模型组(2m1/kg 0.9%氯化钠液腹腔注射+1ml/10Og 75%乙醇灌胃),低剂量ghrelin组(2p,g/kg ghrelin腹腔注射+lml/100g 75%7,醇灌胃),高剂量ghrelin组(20μg/kg ghrelin腹腔注射+lml/100g 75%乙醇灌胃)
2小时后四组大鼠麻醉取胃。
2.计算各组大鼠胃黏膜溃疡指数。
3.HE染色观察胃黏膜炎症损伤程度。
4.通过荧光半定量RT-PCR法测定eNOS/iNOS和COX-1/COX-2的mRNA表达,
实验二
1.64只雌性SD大鼠分成四组,空白对照组(2m1/kg 0.9%氯化钠液腹腔注射+lml/100g 0.9%氯化钠液灌胃),乙醇模型组(2ml/kg 0.9%氯化钠液腹腔注射+1ml/100g 75%乙醇灌胃),ghrelin组(20μg/kg ghrelin腹腔注射+lml/100g 75%7,醇灌胃),Capsazepine组(20μg/kg ghrelin腹腔注射+1mg/kg Capsazepine腹腔注射+1ml/100g 75%乙醇灌胃),2小时后四组大鼠麻醉取胃。
2.计算各组大鼠胃黏膜溃疡指数。
3.HE染色观察胃黏膜炎症损伤程度。
4.通过荧光半定量RT-PCR法测定eNOS(?)(?)iNOS(?)(?)COX-1/COX-2的1nRNA表达,测定CGRP的mRNA的表达。
5.酶联免疫吸附试验(ELISA)法测定胃组织中CGRP含量。
结果
实验一
1. Ghrelin组胃溃疡指数明显小于乙醇模型组,与Ghrelin呈剂量相关性。
2. Ghrelin组病理炎症反应较乙醇模型组明显减轻,与Ghrelin呈剂量相关性。
3.荧光半定量RT-PCR:ghrelin组与乙醇模型组相比,eNOS mRNA表达上升,COX-1 mRNA表达无变化,但iNOS mRNA和COX-2表达下降,与ghrelin呈剂量相关性。
实验二
1. Capsazepine组胃溃疡指数明显大于ghrelin组。
2. Capsazepine组病理炎症反应明显较ghrelin组加重。
3.荧光半定量RT-PCR:capsazepine组与ghrelin组相比,iNOS和COX-2mRNA表达上升；(;NOS(?)(?)CGRP mRNA表达下降,COX-1 mRNA表达无变化
4.酶联免疫吸附试验(ELISA) CGRP测定：capsazepine组与ghrelin组相比,组织中CGRP含量下降。
结论
本实验结果提示Ghrelin具有对乙醇诱导的胃黏膜保护作用,其保护机制可能是通过参与辣椒素一敏感感觉神经保护性反射,调节CGRP的表达从而参与调控NOS和COX活性来达到胃黏膜保护的作用。
Objective
To investigate the gastroprotective effects of ghrelin against ethanol-induced acute gastric injury in rats and possible mechanism.
Methods
Part 1
1.64 female Sprague-Dawley(SD) rats were divided into four groups and were intragastrically administrated as follows:Control group(2mg/kg 0.9%NaCl ip+lml/100g 0.9%NaCl ig);Ethanol model group(2ml/kg 0.9%NaCl ip+lml/100g 75%ethanol ig);Low-dose ghrelin group(2μg/kg ghrelin ip+lml/100g 75%ethanol ig);High-dose ghrelin group(20μg/kg ghrelin ip+lml/100g 75%ethanol ig).Rats were anaesthetized and the stomachs were removed after 2 hours.
2. The ulcer index of gastric mucosal lesions in rats were calculated.
3. Gastric mucosal inflammation were assessed by hematoxylin-eosin (H&E) staining.
4. eNOS/iNOS and COX-1/COX-2 mRNA expression in these specimens were verified by semi-quantitative reverse transcriptional polymerase chain reaction (SqRT-PCR).
Part 2
1.64 female SD rats were divided into four groups and were intragastrically administrated as follows:Control group(2mg/kg 0.9%NaCl ip+lml/100g 0.9%NaCl ig);Ethanol model group(2ml/kg 0.9%NaCl ip+lml/100g 75%ethanol ig);Ghrelin group(2μg/kg ghrelin ip+lml/100g 75%ethanol ig);Capsazepine group(20μg/kg ghrelin ip+lmg/kg Capsazepine ip+lml/100g 75%ethanol ig).Rats were anaesthetized and the stomachs were removed after 2 hours.
2. The ulcer index of gastric mucosal lesions in rats were calculated.
3. The inflammatory lesions of gastric mucosa were assessed by hematoxylin-eosin (H&E) staining.
4. eNOS/iNOS, COX-1/COX-2 and CGRP mRNA expression in these specimens were verified by SqRT-PCR
5.The levels of CGRP in these specimens were measured by Enzyme-linked immunosorbent assay (ELISA).
Results
Part 1
1. The ulcer index of ghrelin groups were significantly less than that of ethanol model group, and were positively correlated with dosages of ghrelin.
2. Compared with ethanol model group,the gastric mucosal inflammation lesions of ghrelin group were significantly reduced, and were positively correlated with dosages of ghrelin.
3. Compared with Ethanol model group,grhrein groups were characterized by no changes of COX-1 mRNA level, but iNOS mRNA and COX-2 mRNA decreased, eNOS mRNA increased in a dose-dependent manner.
Part 2
1. The ulcerative index of capsazepine group were significantly greater than those of ghrelin group.
2. Compared with ghrelin group,the gastric mucosal inflammation lesions of capsazepine group increased significantly.
3. Compared with ghrelin group, there was no change of COX-1 mRNA level in capsazepine group,but mRNA levels of COX-2 and iNOS increased while eNOS and CGRP decreased.
4. Compared with ghrelin group, the levels of CGRP in gastric tissue of capsazepine decreased.
Conclusions
Ghrelin had protective effects on ethanol-induced acute gastric mucosal injury in rats.The mechanism of the protective effects was related to CGRP which can be involved in regulating capsaicin-sensitive nerves and thus participated in the regulation of the activity of NOS and COX.
引文
1. Kojima M,Hosoda H,Date Y,et al.Ghrelin is a growth-hormone-releasing acylated peptide from stomach[J].Nature,1999;402(6762):656-60
2. Hosda H,Kojima M,Matsuo H et al.Ghrelin and des-acyl ghrelin:two major forms of rat ghrelin peptide in gastrointestinal tissue[J].Biochem Biophys Res Commun,2000;279(3):909-13
3. Rindi G,Necchi V,Savio A,et al.Characterisation of gastric ghrelin cell in man and other mammals:studies in adult and fetal tissue[J].Histochem Cell Bio, 2002;117(6):511-9
4. Hosoda H,Kojima M,Mizushima T,et al.Structural divergence of human ghre-lin.Identification of multiple ghrelin-derived molecules produced by post-trans-lational processing[J].Biol Chem,2003;278(1):64-70
5. Gnanapavan S,Kola B,Bustin SA,et al. The tissue distribution of the mRNA of ghrelin and subtypes of its receptor, GHS-R, in humans[J].Clin Endocrinol Metab,2002;87(6):2988-91
6. Papotti M,Ghe C,Cassoni P,et al.Growth hormone secretagogue binding sites in perpheral human tissues[J].Clin Endocrinol Metab,2000;85(10):3803-7.
7. Wren AM,Small CJ,Ward HL,et al.The novel hypothalamic peptide ghrelin stimulates food intake and growth hormone secretion[J].Endocrinology,2000; 141(11):4325-8.
8. Wang L,Saint-Pierre DH,Tache Y.Peripheral ghrelin selectively increases Fos expression in neuropeptide Y-synthesizing neurons in mouse hypothalamic arcuate nucleus[J].Neurosci Lett,2002;325(1):47-51
9. Date Y,Murakami N,Toshinai K,et al.The role of the gastric afferent vagal nerve in ghrelin-induced feeding and growth hormone secretion in rats[J]. Gastroenterology,2002; 123(4):1120-8
10. Masuda Y,Tanaka T,Inomata N,et al.Ghrelin stimulates gastric acid secretion and motility in rats[J].Biochem Biophys Res Commun,2000;276(3):905-8
11.Sakata I,Yamazaki M,Inoue K,et al.Growth hormone secretagogue receptor expression in the cells of the stomach-projected afferent nerve in the rat nodose ganglion[J].Neurosci Lett,2003;342(3):183-6
12. Sibilia V,Rindi G,Pagani F et al.Ghrelin protects against ethanol-induced gastric ulcers in rats:studies on the mechanisms of action[J].Endocrinology, 144(1):353-9
13. Brzozowski T,Konturek PC,Konturek SJ,et al.Exogenous and endogenous Ghrelin in gastroprotection against stress-induced gastric damage[J].Regul Pept,2004;120(1-3):39-51
14. Holzer P,Neural emergency in the stomach[J].Gastroenterology,1998;114(4):823-39
15. Peskar BM.Neural aspects of prostaglandin involvement in gastric mucosal defense[J].Physiol Pharmacol,2001;52(4):555-68
16. Green T,Dockray GJ.Characterization of the peptidergic afferent innervation of the stomach in the rat,mouse and guinea-pig[J].Neuroscience,1988;5(1):181-93
17. Sternini C,Anderson K.Calcitonin gene-related peptide-containing neurons supplying the rat digestive system:differential disteibution and expression pattern[J].Somatosens Mot Res,1992;9(1):45-59
18.贾云丹,陈曦,唐明,蒋正尧.功能性Ghrelin受体在内脏迷走和脊髓传入神经通路中的表达[J].生理学报Acta Physiologica Sinica,2008;60(1):149-55
19.徐叔云,卞如濂,陈修.药理实验方法学[M].第2版.北京：人民卫生出版社,1991；1158-62
20. Sawaoka H,Tsuji S,Tsujii M,et al.Expression of the cyclooxygenase-2 gene in gastric epithelium[J].Clin Gastroenterol,1997;25(1):105-10
21. Brzozowski T,Konturek PC,Drozdowicz D,et al.Role of central and peripheral ghrelin in the mechanism of gastric mucosal defence[J].Inflammopharmaco-logy,2005;13(1-3):45-62
22. Sibilia V,Pagani F,Rindi G,et al.Central ghrelin gastroprotection involves nitric oxide/prostaglandin cross-talk[J].British Journal of Pharmacology,2008; 154(3): 688-97
23. Clemeti G,Caruso A,Prato A,et al.A role of nitric oxide in the anti-ulcer activing of calcitionin gene-related peptide[J].Eur J Pharmacol,1994;256(2):7-8
24. Strecker T,Dux M,Messlinger K.Nitric oxide releases calcitonin-gene-related peptide from rat dura mater encephali promoting increases in meningeal blood flow[J].J Vasc Res,2002;39(6):489-96
1. Kojima M,Hosoda H,Date Y,et al.Ghrelin is a growth-hormone-releasing acylated peptide from stomach[J].Nature,1999;402(6762):656-60
2. Kojima M,Kangawa K.Structure and function of Ghrelin[J].Results Probl Cell Diffe,2008;46:89-115
3. Korbonits M,Goldstone AP,Gueorguiev M,et al. Ghrelin-a hormone with multiple functions[J]. Front Neuroendocrinol,2004;25(1):27-68
4. Sato M,Nakahara K,Goto S,et al.Effects of Ghrelin and des-acyl ghrelin on neurogenesis of the rat fetal spinal cord[J].Biochem Biophys Res Commun, 2006;350(3):598-603
5. Gnanapavan S,Kola B,Bustin SA,et al.The tissue distribution of the mRNA of Ghrelin and subtypes of its receptor,GHS-R,in humans[J].Clin Endocrinol Metab 2002;87(6):2988-91
6. Date Y,Kojima M,Hosoda H,et al.Ghrelin,a novel growth hormone-releasing aeylated pepfide,is synthesized in a distinct endocrine cell type in the gastro-intestinal tracts of mats and humans[J].Endocrinology,2000;141(11):4255-61
7. Rindi G,Necchi V,Savio A,et al.Characterisation of gastric ghrelin cell in man and other mammals:studies in adult and fetal tissue[J].Histochem Cell Bio,2002;117(6):511-9
8. Sakata I,Nakamura K,Yamazaki M,et al.Ghrelin-producing cells exist as two types of cells,closed and opened-type cells,in the rat gastrointestinal tract[J]. Pepfides,2002;23(3):531-6
9. Dornonville de la Cour C,Bjorkqvist M, Sandvik AK,et al.A-like cells in the rat stomach contain ghrelin and do not operate under gastrin control[J]. Regul Pept,2001;99(2-3):141-50
10. Masuda Y,Tanaka T,Inomata N,et al.Ghrelin stimulates gastric acid secretion and motility in rats[J].Biochem Biophys Res Commun,2000;276(3):905-8
11. Depoortere I,Thijs T,Thielemans L,et al.Interaction of the growth hormone-releasing peptides ghrelin hormone-releasing peptide-6 with the motilin recept or in the rabbit gastric antrum[J].Pharmacol Exp Ther,2003;305(2):660-7
12. Trudel L,Tomasetto C,Rio MC,et al.Ghrelin/motilin-related peptide is a potent prokinetic to reverse gastric postoperative ileus in rats[J].Am J Physiol Gastroinest Liver Physiol,2002;282(6):948-52
13. Edholm T,Levin F,Hellstrom PM,et al.Ghrelin stimulates motility in the small intestine of rats through intrinsic cholinergic neurons [J]. Regul Pept,2004;121 (1-3):25-30
14. Sallam HS,Oliveira HM,Gan HT,et al.Ghrelin improves burn-induced delayed gastrointestinal transit in rats[J].Am J Physiol Regul Integr Comp Physiol, 2007;292(1):253-7
15. Fujimiya M,Asakawa A,Fujino K,et al.Acylated ghrelin and des-acyl ghrelin exert different effects on the gastrointestinal motility in conscious rats[J]. Intemational Congress Series,2006;1287:361-7
16. Bassil AK,Dass NB,Sanger GJ.The prokinetic-like activity of ghrelin in rat isolated stomach is mediated via cholinergic and tachykininergic motor neurons[J]. Eur J Pharmacol,2006;544(1-3):146-52
17. Ariga H,Imai K,Chen C,et al.Does ghrelin explain accelerated gastric empty-ing in the early stages of diabetes mellitus?[J].Am J Physiol Regul Integr Comp Physiol,2008;294(6):1807-12
18. Tack J,Depoortere I,Bisschops R,et al.Influence of ghrelin on interdigestive gastrointestinal motility in humans[J].Gut,2006;55(3):327-33
19. Ariga H,Tsukamoto K,Chen C,et al.Endogenous acyl ghrelin is involved in mediating spontaneous phase Ⅲ-like contractions of the rat stomach[J].Neuro-gastroenterol Motil,2007;19(8):675-80
20. Sibilia V,Rindi G,Pagani F et al.Ghrelin protects against ethanol-induced gas tric ulcers in rats:studies on the mechanisms of action[J].Endocrinology, 2003;144(1):353-9
21. Brzozowski T,Konturek PC,Konturek SJ,et al.Exogenous and endogenous ghrelin in gastroprotection against stress-induced gastric damage [J].Regul Pept, 2004; 120(1-3):39-51.
22. Brzozowski T,Konturek PC,Drozdowicz D,et al.Role of central and peripheral ghrelin in the mechanism of gastric mucosal defence[J].Inflammopharmaco-logy,2005;13(1-3):45-62
23. Konturek PC,Brzozowski T,Konturek SJ,et al.Role of histamine in ghrelin-induced gastroprotection against acute gastric lesions[J].Inflamm Res,2007;56(1):S25-6
24. Nwokolo CU,Freshwater DA,O'Hare P,et al.Plasma ghrelin following cure of Helicobacter pylori[J].Gut,2003;52(5):637-40
25. Gokcel A,Gumurdulu Y,Kayaselcuk F,et al.Helicobacter pylori has no effact on plasma ghrelin levels[J].Eur J Endocrinol,2003;148(4):423-6
26. Osawa H,Nakazato M, Date,Y,et al.Impaired production of gastric Ghrelin in chronic gastritis associated with Helicobacter pylori[J].Clin Endocrinol Metab, 2005;90(1):10-6
27. D'Onghia V,Leoncini R,Carli R,et al. Circulating gastrin and ghrelin levels in patients with colorectal cancer:correlation with tumour stage, Helicobacter pylori infection and BMI[J].Biomed Pharmacother,2007;61(2-3):137-41
28. Waseem T, Javaid-ur-Rehman,Ahmad F,et al. Role of ghrelin axis in colorectal cancer:a novel association[J].Peptides,2008;29(8):1369-76
29. Mark S,Talat W,Hiromichi I,et al.Ghrelin promotes pancreatic adenocarcinoma cellular proliferation and invasiveness[J].Biochem Biophys Res Commun,2003; 309(2):464-8
30. Lee HM,Wang G,Englander EW,et al.Ghrelin,a new gastrointestinal endocrine peptide that stimulates insulin secretion:enteric distribution,ontogeny,influence of endocrine,and dietary manipulations[J].Endocrinology,2002; 143(1):185-190
31. 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):124-9
32. 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):241-4
2. Hosda H,Kojima M,Matsuo H et al.Ghrelin and des-acyl ghrelin:two major forms of rat ghrelin peptide in gastrointestinal tissue[J].Biochem Biophys Res Commun,2000;279(3):909-13
3. Rindi G,Necchi V,Savio A,et al.Characterisation of gastric ghrelin cell in man and other mammals:studies in adult and fetal tissue[J].Histochem Cell Bio, 2002;117(6):511-9
4. Hosoda H,Kojima M,Mizushima T,et al.Structural divergence of human ghre-lin.Identification of multiple ghrelin-derived molecules produced by post-trans-lational processing[J].Biol Chem,2003;278(1):64-70
5. Gnanapavan S,Kola B,Bustin SA,et al. The tissue distribution of the mRNA of ghrelin and subtypes of its receptor, GHS-R, in humans[J].Clin Endocrinol Metab,2002;87(6):2988-91
6. Papotti M,Ghe C,Cassoni P,et al.Growth hormone secretagogue binding sites in perpheral human tissues[J].Clin Endocrinol Metab,2000;85(10):3803-7.
7. Wren AM,Small CJ,Ward HL,et al.The novel hypothalamic peptide ghrelin stimulates food intake and growth hormone secretion[J].Endocrinology,2000; 141(11):4325-8.
8. Wang L,Saint-Pierre DH,Tache Y.Peripheral ghrelin selectively increases Fos expression in neuropeptide Y-synthesizing neurons in mouse hypothalamic arcuate nucleus[J].Neurosci Lett,2002;325(1):47-51
9. Date Y,Murakami N,Toshinai K,et al.The role of the gastric afferent vagal nerve in ghrelin-induced feeding and growth hormone secretion in rats[J]. Gastroenterology,2002; 123(4):1120-8
10. Masuda Y,Tanaka T,Inomata N,et al.Ghrelin stimulates gastric acid secretion and motility in rats[J].Biochem Biophys Res Commun,2000;276(3):905-8
11.Sakata I,Yamazaki M,Inoue K,et al.Growth hormone secretagogue receptor expression in the cells of the stomach-projected afferent nerve in the rat nodose ganglion[J].Neurosci Lett,2003;342(3):183-6
12. Sibilia V,Rindi G,Pagani F et al.Ghrelin protects against ethanol-induced gastric ulcers in rats:studies on the mechanisms of action[J].Endocrinology, 144(1):353-9
13. Brzozowski T,Konturek PC,Konturek SJ,et al.Exogenous and endogenous Ghrelin in gastroprotection against stress-induced gastric damage[J].Regul Pept,2004;120(1-3):39-51
14. Holzer P,Neural emergency in the stomach[J].Gastroenterology,1998;114(4):823-39
15. Peskar BM.Neural aspects of prostaglandin involvement in gastric mucosal defense[J].Physiol Pharmacol,2001;52(4):555-68
16. Green T,Dockray GJ.Characterization of the peptidergic afferent innervation of the stomach in the rat,mouse and guinea-pig[J].Neuroscience,1988;5(1):181-93
17. Sternini C,Anderson K.Calcitonin gene-related peptide-containing neurons supplying the rat digestive system:differential disteibution and expression pattern[J].Somatosens Mot Res,1992;9(1):45-59
18.贾云丹,陈曦,唐明,蒋正尧.功能性Ghrelin受体在内脏迷走和脊髓传入神经通路中的表达[J].生理学报Acta Physiologica Sinica,2008;60(1):149-55
19.徐叔云,卞如濂,陈修.药理实验方法学[M].第2版.北京：人民卫生出版社,1991；1158-62
20. Sawaoka H,Tsuji S,Tsujii M,et al.Expression of the cyclooxygenase-2 gene in gastric epithelium[J].Clin Gastroenterol,1997;25(1):105-10
21. Brzozowski T,Konturek PC,Drozdowicz D,et al.Role of central and peripheral ghrelin in the mechanism of gastric mucosal defence[J].Inflammopharmaco-logy,2005;13(1-3):45-62
22. Sibilia V,Pagani F,Rindi G,et al.Central ghrelin gastroprotection involves nitric oxide/prostaglandin cross-talk[J].British Journal of Pharmacology,2008; 154(3): 688-97
23. Clemeti G,Caruso A,Prato A,et al.A role of nitric oxide in the anti-ulcer activing of calcitionin gene-related peptide[J].Eur J Pharmacol,1994;256(2):7-8
24. Strecker T,Dux M,Messlinger K.Nitric oxide releases calcitonin-gene-related peptide from rat dura mater encephali promoting increases in meningeal blood flow[J].J Vasc Res,2002;39(6):489-96
1. Kojima M,Hosoda H,Date Y,et al.Ghrelin is a growth-hormone-releasing acylated peptide from stomach[J].Nature,1999;402(6762):656-60
2. Kojima M,Kangawa K.Structure and function of Ghrelin[J].Results Probl Cell Diffe,2008;46:89-115
3. Korbonits M,Goldstone AP,Gueorguiev M,et al. Ghrelin-a hormone with multiple functions[J]. Front Neuroendocrinol,2004;25(1):27-68
4. Sato M,Nakahara K,Goto S,et al.Effects of Ghrelin and des-acyl ghrelin on neurogenesis of the rat fetal spinal cord[J].Biochem Biophys Res Commun, 2006;350(3):598-603
5. Gnanapavan S,Kola B,Bustin SA,et al.The tissue distribution of the mRNA of Ghrelin and subtypes of its receptor,GHS-R,in humans[J].Clin Endocrinol Metab 2002;87(6):2988-91
6. Date Y,Kojima M,Hosoda H,et al.Ghrelin,a novel growth hormone-releasing aeylated pepfide,is synthesized in a distinct endocrine cell type in the gastro-intestinal tracts of mats and humans[J].Endocrinology,2000;141(11):4255-61
7. Rindi G,Necchi V,Savio A,et al.Characterisation of gastric ghrelin cell in man and other mammals:studies in adult and fetal tissue[J].Histochem Cell Bio,2002;117(6):511-9
8. Sakata I,Nakamura K,Yamazaki M,et al.Ghrelin-producing cells exist as two types of cells,closed and opened-type cells,in the rat gastrointestinal tract[J]. Pepfides,2002;23(3):531-6
9. Dornonville de la Cour C,Bjorkqvist M, Sandvik AK,et al.A-like cells in the rat stomach contain ghrelin and do not operate under gastrin control[J]. Regul Pept,2001;99(2-3):141-50
10. Masuda Y,Tanaka T,Inomata N,et al.Ghrelin stimulates gastric acid secretion and motility in rats[J].Biochem Biophys Res Commun,2000;276(3):905-8
11. Depoortere I,Thijs T,Thielemans L,et al.Interaction of the growth hormone-releasing peptides ghrelin hormone-releasing peptide-6 with the motilin recept or in the rabbit gastric antrum[J].Pharmacol Exp Ther,2003;305(2):660-7
12. Trudel L,Tomasetto C,Rio MC,et al.Ghrelin/motilin-related peptide is a potent prokinetic to reverse gastric postoperative ileus in rats[J].Am J Physiol Gastroinest Liver Physiol,2002;282(6):948-52
13. Edholm T,Levin F,Hellstrom PM,et al.Ghrelin stimulates motility in the small intestine of rats through intrinsic cholinergic neurons [J]. Regul Pept,2004;121 (1-3):25-30
14. Sallam HS,Oliveira HM,Gan HT,et al.Ghrelin improves burn-induced delayed gastrointestinal transit in rats[J].Am J Physiol Regul Integr Comp Physiol, 2007;292(1):253-7
15. Fujimiya M,Asakawa A,Fujino K,et al.Acylated ghrelin and des-acyl ghrelin exert different effects on the gastrointestinal motility in conscious rats[J]. Intemational Congress Series,2006;1287:361-7
16. Bassil AK,Dass NB,Sanger GJ.The prokinetic-like activity of ghrelin in rat isolated stomach is mediated via cholinergic and tachykininergic motor neurons[J]. Eur J Pharmacol,2006;544(1-3):146-52
17. Ariga H,Imai K,Chen C,et al.Does ghrelin explain accelerated gastric empty-ing in the early stages of diabetes mellitus?[J].Am J Physiol Regul Integr Comp Physiol,2008;294(6):1807-12
18. Tack J,Depoortere I,Bisschops R,et al.Influence of ghrelin on interdigestive gastrointestinal motility in humans[J].Gut,2006;55(3):327-33
19. Ariga H,Tsukamoto K,Chen C,et al.Endogenous acyl ghrelin is involved in mediating spontaneous phase Ⅲ-like contractions of the rat stomach[J].Neuro-gastroenterol Motil,2007;19(8):675-80
20. Sibilia V,Rindi G,Pagani F et al.Ghrelin protects against ethanol-induced gas tric ulcers in rats:studies on the mechanisms of action[J].Endocrinology, 2003;144(1):353-9
21. Brzozowski T,Konturek PC,Konturek SJ,et al.Exogenous and endogenous ghrelin in gastroprotection against stress-induced gastric damage [J].Regul Pept, 2004; 120(1-3):39-51.
22. Brzozowski T,Konturek PC,Drozdowicz D,et al.Role of central and peripheral ghrelin in the mechanism of gastric mucosal defence[J].Inflammopharmaco-logy,2005;13(1-3):45-62
23. Konturek PC,Brzozowski T,Konturek SJ,et al.Role of histamine in ghrelin-induced gastroprotection against acute gastric lesions[J].Inflamm Res,2007;56(1):S25-6
24. Nwokolo CU,Freshwater DA,O'Hare P,et al.Plasma ghrelin following cure of Helicobacter pylori[J].Gut,2003;52(5):637-40
25. Gokcel A,Gumurdulu Y,Kayaselcuk F,et al.Helicobacter pylori has no effact on plasma ghrelin levels[J].Eur J Endocrinol,2003;148(4):423-6
26. Osawa H,Nakazato M, Date,Y,et al.Impaired production of gastric Ghrelin in chronic gastritis associated with Helicobacter pylori[J].Clin Endocrinol Metab, 2005;90(1):10-6
27. D'Onghia V,Leoncini R,Carli R,et al. Circulating gastrin and ghrelin levels in patients with colorectal cancer:correlation with tumour stage, Helicobacter pylori infection and BMI[J].Biomed Pharmacother,2007;61(2-3):137-41
28. Waseem T, Javaid-ur-Rehman,Ahmad F,et al. Role of ghrelin axis in colorectal cancer:a novel association[J].Peptides,2008;29(8):1369-76
29. Mark S,Talat W,Hiromichi I,et al.Ghrelin promotes pancreatic adenocarcinoma cellular proliferation and invasiveness[J].Biochem Biophys Res Commun,2003; 309(2):464-8
30. Lee HM,Wang G,Englander EW,et al.Ghrelin,a new gastrointestinal endocrine peptide that stimulates insulin secretion:enteric distribution,ontogeny,influence of endocrine,and dietary manipulations[J].Endocrinology,2002; 143(1):185-190
31. 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):124-9
32. 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):241-4