摘要
目的探讨不同浓度的草药复方制剂STW-5-NⅡ对不同化学物及机械刺激诱导的大鼠小肠传入神经电活动变化的影响,并分析其可能的机制。
方法24只雄性Wister大鼠均分为4组,动物麻醉后经胃管灌入不同浓度STW-5-NⅡ药液(原液、50%原液、25%原液)及对照液。用细胞外多元肠系膜传入神经记录技术(extracellular multi-unit mesenteric afferent nerverecording)描记肠传入神经元电活动信号。给予不同剂量5-羟色胺(5-HT)(5,10,20,40μg/kg)或不同剂量缓激肽(BK)(15,30,60μg/kg)静脉注射,以及不同程度的肠管扩张(0cmH_2O～60cmH_2O)刺激,观察各种刺激前后小肠传入神经电活动的变化,比较和分析不同浓度STW-5-NⅡ对此变化的影响。
结果STW-5-NⅡ对小肠传入神经基本电活动没有影响。不同剂量的5-HT和BK均可引起小肠传入神经电活动的增加;中、高浓度的STW-5-NⅡ明显降低BK的这种增加作用(P＜0.05,或P＜0.01);肠管扩张刺激在20-60cmH_2O时小肠传入神经电活动增加,不同浓度的STW-5-NⅡ对此有降低作用,对5-HT诱导的小肠传入神经电活动的增加也有降低的趋势,但作用不明显(P＞0.05)。
结论STW-5-NⅡ可以降低大鼠小肠对某些化学刺激和机械刺激的敏感性,可能有助于与内脏敏感性增高有关的胃肠道运动功能障碍性疾病的治疗。
目的:研究大麻素受体激动剂、拮抗剂对高敏感小肠传入神经电活动的影响,对大鼠小肠平滑肌电活动的影响,以探讨内源性大麻素系统对胃肠运动的作用。
方法:离体试验:将雄性Wistar大鼠麻醉后取空肠肠管,用细胞外多元肠系膜传入神经记录技术(extracellular multi-unit mesenteric afferent nerve recording)描记肠传入神经元电活动信号。然后肠管灌流大麻素受体激动剂(Win55,10~(-5)mol/L)或拮抗剂(SR141716A,10~(-5)mol/L),给予5-羟色胺(5-HT,500μM)、缓激肽(BK,1μM)或不同程度的肠管扩张(0cmH_2O～80cmH_2O)刺激,观察各刺激前后小肠传入神经电活动的变化。在体试验:用PowerLab微电极法,描记大鼠胃肠平滑肌电活动,大麻素受体激动剂(HU210,100μg/kg)或拮抗剂(AM251,3mg/kg)腹腔注射,观察大鼠胃肠平滑肌电活动的变化。
结果:大麻素受体拮抗剂SR141716A(10~(-5)mol/L)明显降低肠管扩张(30cmH_2O～80cmH_2O)以及BK(1μM)诱导的离体小肠传入神经电活动的增加(P＜0.05),而大麻素受体激动剂Win55对此类化学、机械刺激诱发的小肠传入神经电活动无明显影响;SR141716A的类似物AM251(3mg/kg)明显增强在体大鼠小肠平滑肌峰电活动的频率及幅度,而大麻素受体激动剂HU210(100μg/kg)则明显降低在体大鼠小肠平滑肌峰电活动的频率及幅度。
结论:内源性大麻素系统参与大鼠胃肠运动的调节。在整体状态下,该系统对大鼠小肠平滑肌峰电活动起抑制作用;对离体肠管,大麻素受体拮抗剂以及大麻素受体激动剂对其传入神经基础电活动没有明显影响,但某些病因引起的肠管传入神经电活动的增加(高敏感状态)受大麻素受体拮抗剂所抑制,提示局部大麻素系统活动增强可能参与了肠道高敏感的发生。
Objective:To investigate the effect of different concentration of a new simplified herbal combination product STW-5-NII on intestinal afferent nerve discharge after different mechanical and chemical stimuli.
Methods:24 male Wister Rats were divided into four groups.STW-5-NII at a concentration of 100%,50%,25%or vehicle were administered orally in rats after anesthetized.The extracellular multi-unit mesenteric afferent nerve recordings were obtained in the proximal jejunum.Afferent discharge to 5-Hydroxytryptamine(5-HT) (5、10、20 and 40μg kg~(-1),i.v.),bradykinin(BK)(15、30 and 60μg kg~(-1)i.v.)and luminal distension(0cmH_2O -60cmH_2O)were recorded and analyzed.
Results:Afferent nerve discharge was increased after different dose of 5-HT and BK. Afferent nerve sensitivity to bradykinin but not to 5-HT was significantly different depending on the dose of pretreatment with STW-5-NII.A pressure-dependent increase in afferent nerve discharge was observed during intraluminal distension that was not significant different following pretreatment with any concentraction of STW-5-NII.
Conclusions:The herbal preparation STW-5-NII desensitized intestinal afferents selectively to systemic bradykinin.This new preparation might be helpful for patients suffering from disorders associated with visceral hypersensitivity.
Objective To investigate the effects of CB agonist and antagonist on the afferent nerve discharge in the intestinal experimental hypersensitivity and the intestinal myoelectric activity in rats.
Methods Experiment in vitro:Wistar rats were anesthetized and ileal segments removed with the mesenteric arcade attached.The segments were perfused with Krebs' buffer(37℃)which was either supplemented with the CB-1 receptor agonist WIN 55,212-2(10~(-5)mold),or the CB-1 receptor antagonist SR 141716A(10~(-5)mol/l), or vehicle,respectively,(n=6,each).The changes of intestinal afferent discharge were observed after adding 5-HT(500μM)or BK(1μM)or luminal distension(0 cmH_2O～80cmH_2O).The afferent nerve discharge was recorded using extracellular multi-unit mesenteric afferent nerve recording.Experiment in vivo:SD rat intestinal myoelectric motility was recorded using PowerLab system(Australia),and the effect of CB receptor agonist HU210(100μg/kg)or CB receptor antagonist AM 251(3mg/kg)was observed after their intraperitoneal injection respectively(n=6, each).
Results in vitro:Using CB-1 receptor antagonist SR141716A,the intestinal afferent nerve discharge induced by bradykinin stimulation was reduced significantly (P＜0.05),and the same effect was also found in the afferent nerve discharge induced by the noxious mechanical distension of the intestinal loop(30 to 80cmH_2O) (P＜0.05).in vivo:CB receptor antagonist AM 251(3mg/kg)increased the frequencies and amplitudes of the intestinal spike activities in rats.While CB receptor agonist HU210(100μg/kg)got the oppositive effection on the frequencies and amplitudes of the intestinal myoelectric activities in rats.
Conclusion The endocannabinoid system took part in the regulation of the gastrointestinal motility.This system seemed to play an inhibiting role in the intestinal myoelectric activity in rats.It had no effect on the baseline of the afferent discharge from the isolated intestinal segments of rats.But the increment of the afferent discharge induced by some stimulatiors was attenuated by the CB antagonist significantly,which suggested that the increased activity of the local endocannabinoid system may be involved in the pathogenesis of the small intestinal hypersensitivity.
引文
1.梁列新,候晓华.中华实用内科学杂志.2004,24(8):511-512
2.张敏敏,李兆申.肠道迷走神经感觉通路调节机制的研究进展.胃肠病学,2002,7(5):315.
3.Barbara G,De Giorgio R,Stanghellini V,et al.A role for inflammation in irritable bowel syndrome.Gut,2002,51(Suppl 1):i41-i44.
4.Collins SM,V alliance B,Barbara G,et al.Putative inflammatory and immunological mechanisms in functional bowel disorders.Baillieres Best Pract Res Clin Gastroenterol,1999,13(3):429-436.
5.Melzer J,Rosch W,Reichling J,et al.Meta-analysis:phytotherapy of functional dyspepsia with the herbal drug preparation STW 5(Iberogast).Aliment Pharmacol Ther,2004,20:1279-1287.
6.Rosch W,Liebregts T,Gundermann KJ,et al.Phytotherapy for functional dyspepsia:A review of the clinical evidence for the herbal preparation STW 5.Phytomedicine,2006,13(Suppl)1:114-121.
7.Allescher HD.Functional dyspepsia-A multicausal disease and its therapy.Phytomedicine,2006,13(Suppl1):2-11.
8.Madisch A,Melderis H,Mayr G,et al.A plant extract and its modified preparation in functional dyspepsia.Results of a double-blind placebo controlled comparative study.Z Gastroenterol,2001,39:511-517.
9.Liu CY,Mueller MH,Glatzie J,et al.The herbal preparation STW 5(Iberogast)desensitizes intestinal afferents in the rat small intestine.Neurogastroenterol Motil,2004,16:759-764.
10.Pertwee RG.Cannabinoid receptors and pain.Prog Neurobiol 2001,63:569-611.
11.Di Marzo V,Bifulco M,De Petrocellis L.Endocannabinoids and multiple sclerosis:a blessing from the 'inner bliss'.Trends Pharmacol Sci 2000,21:195-197.
12.Curatolo M,Bogduk N.Pharmacological pain treatment of musculoskeletal disorders:current perspectives and future prospects.Clin J Pain 2001,17:25-32.
13.De Petrocellis L,Melck D,Bisogno T,et al.Endocannabinoids and fatty acid amides in cancer,inflammation and related disorders.Chem Phys Lipids 2000,108:191-209.
14.Grispoon L,Bakalar JB.Marihuana.The forbidden medicine.New Haven,CT:Yale University Press;1997
15.Razden RK.Structure-activity relation in cannabinoids.Pharmacol Rew,1986,38:175-149.
16.Pertwee RG.Cannabinoid receptor ligands:clinical and neuropharmacological considerations relevant to future drug discovery and development.Expert Opin Invest Drugs 2000,9:1553-1571.
17. Devane WA, Hanus L, Breuer A, et al. Isolation and structure of a brain constituent that binds to the cannabinoid receptor. Science, 1992,258 : 1946 - 1949.
18. Izzo AA, Fezza F, Capasso R, et al. Cannabinoid CB(1)-receptor mediated regulation of gastrointestinal motility in mice in a model of intestinal inflammation.Br J Pharmacol 2001,134:563 - 570
19. Wilson RI, Nicoll RA. Endocannabinoid signaling in the brain. Science, 2002,296(5568):678-682
20. Katayama K, Ueda N, Kurahashi Y, et al. Distribution of anandamide amidohydrolase in rat tissues with special reference to small intestine. Biochim Biophys Acta, 1997,1347:212-218
21. Pinto L, Izzo AA, Cascio MG, et al. Endocannabinoids as physiological regulators of colonic propulsion in mice. Gastroenterology, 2002,123:227-234
22. Pertwee RG, Fernando SR, Griffin G, et al. Effect of phenylmethylsulphonyl fluoride on the potency of anandamide as an inhibitor of electrically evoked contractions in two isolated tissue preparations. Eur J Pharmacol 1995,272:73 - 78
23. Matsuda LA, Lolait SJ, Brownstein MJ, et al. Structure of a cannabinoid receptor and a functional expression of the cloned cDNA. Nature, 1990,346:561-564.
24. Munro S, Thomas KL, Abu-shaa M, et al. Molecular characterization of a peripheral receptor for cannabinoids. Nature, 1993 ,356 : 61-65.
25. Griffin G, Fernando SR, Ross RA, et al. Evidence for the presence of CB2-like cannabinoid receptors on peripheral nerve terminals. Eur J Pharmacol 1997,339:53-61.
26. Shire D, Carillon C, Kaghad M, et al. An amino-terminal variant of the central cannabinoid receptor resulting from alternative splicing. J Biol Chem 1995, 270:3726-3731.
27. Buckley NE, Hansson S, Harta G,et al. Expression of the CB1 and CB2 receptor messenger RNAs during embryonic development in the rat. Neuroscience 1998, 82:1131-1149.
28. Coutts AA, Irving AJ, Mackie K,et al. Localisation of cannabinoid CB(1) receptor immunoreactivity in the guinea pig and rat myenteric plexus. J Compc Neurol,2002, 448:410-422
29. Pinto L, Capasso R, Di Carlo G,et al. Endocannabinoids and the gut. Prostaglandins Leukot Essent Fatty Acids,2002,66:333-341
30. Coutts AA, Pertwee RG. Evidence that cannabinoidinduced inhibition of electrically evoked contractions of the myenteric plexus-longitudinal muscle preparation of guinea pig small intestine can be modulated by Ca2+ and cAMP. Can J Physiol Pharmacol, 1998, 76:340-346
31. Mascolo N, Izzo AA, Ligresti A, et al. The endocannabinoid system and the molecular basis of paralytic ileus in mice. FASEB J ,2002,16:1973-1975
32. Feinle-Bisset C, Vozzo R, Horowitz M, et al. Diet, food intake, and disturbed physiology in the pathogenesis of symptoms in functional dyspepsia .Am J Gastroenterol, 2004,99:170 -181.
33. Rosemore JG, Lacy BE. Irritable bowel syndrome: basis of clinical management strategies. J Clin Gastroenterol,2002,35(Suppl):37-44.
34.Olden KW.Diagnosis of irritable bowel syndrome.Gastroenterology 2002,122:1701-1714.
35.Pare P.Systematic approach toward the clinical diagnosis of functional dyspepsia.Can J Gastroenterol,1999,13:647-654.
36.时乐,卜平.内脏感觉的检测方法.世界华人消化杂志.2006,14(3):324-331.
37.Melzer J,Rosch W,Reichling J,et al.Meta-analysis:phytotherapy of functional dyspepsia with the herbal drug preparation STW 5(Iberogast).Aliment Pharmacol Ther,2004,20:1279-1287.
38.Kreis ME,Haupt W,Krikup AJ,et al.Histamine sensitivity of mesenteric afferent nerves in the rat jejunum.Am J Physiol Gastrointest Liver Physiol,1998,275(4):675-680.
39.周吕,柯美云.神经胃肠病学与动力—基础与临床.北京:科学出版社,第一版,2005.
40.崔香淑,李在琉.胃肠感觉的神经解剖学基础.诊断学理论与实践,2006,5(1):5-7.
41.Grundy D.Sensory signals from the gastrointestinal tract.Journal of Pediatric Gastroenterology and Nutrition,2005,41(Supply)7-9.
42.Kreis ME,Jiang W,Kirkup AJ,et al.Cosensitivity of vagal mucosal afferents to histamine and 5-HT in the rat jejunum.Am J Physiol Gastrointest Liver Physiol,2002,283:G612-617.
43.Maubach KA,Grundy D.The role of prostaglandins in the bradykinin-induced activation of serosal afferents of the rat jejunum in vitro.J Physiol,1999,515:277-285.
44.Grundy D.Neuroanatomy of visceral nociception:vagal and splanchnic afferent.Gut.2005,51(Suppl Ⅰ):2-5.
45.Krowicki ZK,Moerscbaecher JM,Winsauer PJ,Divagalli SV,Horuby PJ:Δ9-Tetrahydrocannabinol inhibits gastric motility in the rat through cannabinoid CB1receptors.Eur J Pharmacol 1999,37:187-196.
46.Izzo AA,Addeo D,Amato M,et al.In vivo modulation of gastrointestinal motility by cannabinoid drugs.Pharm Pharmacol Commun 2000,6:255-258.
47.Adami M,Bertini S,Frati P,et al.Cannabinoid CB1 receptors are involved in the regulation of rat gastric acid secretion.Pharm Pharmacol Commun 2000,6:273-275.
48.Coruzzi G,Adami M,Coppelli G,et al.Inhibitory effect of the cannabinoid receptor agonist WIN 55,212-2 on pentagastrin-induced gastric acid secretion in the anaesthetized rat.Naunyn Schmiedebergs Arch Pharmacol 1999,360:715-718.
49.Tyler K,Hillard CJ,Greenwood-Van Meerveld B.Inhibition of small intestinal secretion by cannabinoids is CB1 receptor-mediated in rats.Eur J Pharmacol 2000,409:207-211.
50.Izzo AA,Mascolo N,Capasso R,et al.Inhibitory effect of cannabinoid agonists on gastric emptying in the rat.Naunyn Schmiedebergs Arch Pharmacol 1999,360:221-223.
51.Pertwee RG,Fernando SR,Nash JE,et al.Further evidence for the presence of cannabinoid CB1 receptors in guinea-pig small intestine.Br J Pharmacol 1996,118:2199-2205.
52.Izzo AA,Mascolo N,Tonini M,Capasso F.Modulation of peristalsis by cannabinoid CB1 ligands in the isolated guinea-pig ileum.Br J Pharmacol 2000,129:984-990.
53.江澄川,赵志奇,蒋豪主编.疼痛的基础与临床.上海:复旦大学出版社和上海医科大学出版社.2001
54.Poitras P,R iberdy Po itras M,P lourde V,et al.Evolution of visceral sensitivity in patients with irritable bowel syndrome.Dig Dis Sci,2002,47(4):914-920.
55.lzzo AA,Mascolo N,Borrelli F,et al.Excitatory transmission to the circular muscle of the guinea-pig ileum:evidence for the involvement of cannabinoid CB1 receptors.Br J Pharmacol 1998,124:1363-1368.
56.Coutts AA,Pertwee RG:Inhibition by cannabinoid receptor agonists of acetylcholine release from the guinea-pig myenteric plexus.Br J Pharmacol 1997,121:1557-1566.
57.Izzo AA,Pinto L,Borrelli F,et al.Central and peripheral cannabinoid modulation of gastrointestinal transit in physiological states or during the diarrhoea induced by croton oil.Br J Pharmacol 2000,129:1627-1632.
58.Izzo AA,Mascolo N,Pinto L,et al.The role of cannabinoid receptors in intestinal motility,defaecation and diarrhoea in rats.Eur J Pharmacol 1999,384:37-42
59.M.Stoor,A.Sibaev,G.Marsicano,et al.Cannabinoid receptor type1 modulates excitatory and inhibitory neurotransmission in mouse colon.Am J Physiol Gastrointest Liver Physiol 2004286:G110-G117
60.Adami M,Bertini S,Frati P,et al.Cannabinoid CB1 receptors are involved in the regulation of rat gastric acid secretion.Pharm Pharmacol Commun 2000,6:273-275.
61.Federico Massa,Giovanni Marsicano,Heike Hermann,et al.The endogenous cannabinoid system protects against colonic inflammation.J of Clinical Investigation.2004,13(8):1202-1209
62.茹立强.消化管壁内神经丛及其神经元.现代科技综述大辞典(朱永和主编).北京:北京出版社,1998.1301-1302
