足底电击法诱导大鼠内脏高敏感模型的建立和评价
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摘要
目的通过足底电击建立大鼠内脏高敏感动物模型,并评价其有效性和优势。方法将雌性SD大鼠随机分为正常对照(NC)组、足底电击(FSS)组和避水应激(WAS)组。NC组每天放入模拟应激箱中1 h不予刺激;FSS组每天置于电击箱中1 h,给予电击刺激,刺激电压为40 V,刺激频率为20次/min,每次1 s,共5 min;WAS组每天给予1 h避水应激;造模期为10 d。观察记录大鼠一般情况和排便情况,用腹壁回缩反射(AWR)检测内脏敏感性,用ELISA法检测血清CRF、ACTH、CORT、5-HT浓度和结肠组织5-HT浓度。结果 (1) FSS组大鼠的内脏敏感性明显高于WAS组和NC组(P<0.05),结肠黏膜无明显病理性损伤。(2) FSS组大鼠血清CRF、ACTH、CORT、5-HT浓度和结肠组织5-HT浓度均较WAS组和NC组明显升高(P<0.05)。(3) FSS组大鼠排便量明显高于WAS组和NC组(P<0.05),其粪便含水量明显高于NC组(P<0.05),而且其大鼠排便量和粪便含水量数值稳定。结论足底电击造模法可建立大鼠内脏高敏感模型且优于避水应激法。
Objective To establish and evaluate a new rat model of visceral hypersensitivity(VH) induced by electric footshock. Methods Female SD rats were divided randomly into the normal control(NC) group, footshock stress(FSS) group, and water avoidance stress(WAS) group. The NC rats were placed in a footshock stress box for 1 h every day without stimulation. The FSS rats were placed in a footshock stress box for 1 h every day and stimulated by electric shock(U=40 v, t=1 s, f=20 times/min) for 5 min, and WAS rats were placed in a WAS box for 1 h every day. All groups were treated for consecutive 10 days. The general status and defecation status of the rats were recorded every other day. Visceral sensitivity was evaluated with abdominal withdrawal reflex(AWR) test. The serum concentrations of CRF, ACTH, CORT, and 5-HT and the colonic concentration of 5-HT were measured by ELISA. Results 1) Compared with the WAS and NC rats, the visceral sensitivity of FSS rats was significantly increased(P<0.05). There were no significant pathological manifestations in the colons of FSS or WAS rats.(2) Compared with the WAS and NC rats, the serum concentrations of CRF, ACTH, CORT, and 5-HT and the colonic concentration of 5-HT were all significantly increased in the FSS rats(P<0.05).(3) The defecation quantity of FSS rats was significantly greater than those of WAS and NC rats(P<0.05). The water content of the feces of FSS rats was significantly higher than that of NC rats(P<0.05). The defecation quantity and water content in the feces of FSS rats were stable during the experiment. Conclusions FSS is an optimal method for establishing a rat model of VH and is superior to water avoidance stress.
Objective To establish and evaluate a new rat model of visceral hypersensitivity(VH) induced by electric footshock. Methods Female SD rats were divided randomly into the normal control(NC) group, footshock stress(FSS) group, and water avoidance stress(WAS) group. The NC rats were placed in a footshock stress box for 1 h every day without stimulation. The FSS rats were placed in a footshock stress box for 1 h every day and stimulated by electric shock(U=40 v, t=1 s, f=20 times/min) for 5 min, and WAS rats were placed in a WAS box for 1 h every day. All groups were treated for consecutive 10 days. The general status and defecation status of the rats were recorded every other day. Visceral sensitivity was evaluated with abdominal withdrawal reflex(AWR) test. The serum concentrations of CRF, ACTH, CORT, and 5-HT and the colonic concentration of 5-HT were measured by ELISA. Results 1) Compared with the WAS and NC rats, the visceral sensitivity of FSS rats was significantly increased(P<0.05). There were no significant pathological manifestations in the colons of FSS or WAS rats.(2) Compared with the WAS and NC rats, the serum concentrations of CRF, ACTH, CORT, and 5-HT and the colonic concentration of 5-HT were all significantly increased in the FSS rats(P<0.05).(3) The defecation quantity of FSS rats was significantly greater than those of WAS and NC rats(P<0.05). The water content of the feces of FSS rats was significantly higher than that of NC rats(P<0.05). The defecation quantity and water content in the feces of FSS rats were stable during the experiment. Conclusions FSS is an optimal method for establishing a rat model of VH and is superior to water avoidance stress.
引文
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[11] 张北华,王微,王凤云,等,基于不同精神心理应激建立IBS-D大鼠模型的比较研究[J]. 世界中医药,2017(09):2128-2133.Zhang BH, Wang W, Wang FY, et al. Comparative study of establishment of a rat model of IBS-D based on different psychological stress [J]. World Chin Med. 2017(09): 2128-2133.
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[14] Kanada Y, Katayama A, Ikemoto H, et al. Inhibitory effect of the Kampo medicinal formula Yokukansan on acute stress-induced defecation in rats [J]. Neuropsychiatr Dis Treat, 2018, 14: 937-944.
[15] 刘晓萌,张荣,董世芬,等.乌灵菌粉贯叶连翘复方制剂干预对慢性不可预知应激大鼠模型抗抑郁作用及机制的研究[J]. 中国比较医学杂志,2016, 26(05):81-86.Liu XM,,Zhang R, Dong SF, et al. Effects of mixture of Wuling powder and Hypericum perforatum L. on depressed model rats induced by chronic unpredictable stress [J]. Chin J Comp Med, 2016, 26(05): 81-86.
[16] 郭维,孙琪,桂牧微,等.慢性应激损伤对消化系统相关血清肿瘤标志物的影响 [J]. 广东医学,2017(23):3558-3560.Guo W,Sun Q,Gui MW,et al. Experimental study on the effect of chronic stress damage on serum tumor markers [J].Guangdong Med J 2017(23): 3558-3560.
[17] Smith SM, Vale WW. The role of the hypothalamic-pituitary-adrenal axis in neuroendocrine responses to stress [J]. Dialogues Clin Neurosci, 2006, 8(4): 383-395.
[18] Nozu T, Okumura T. Corticotropin-releasing factor receptor type 1 and type 2 interaction in irritable bowel syndrome [J]. J Gastroenterol, 2015, 50(8): 819-830.
[19] Zhou XP, Sha J, Huang L, et al. Nesfatin-1/NUCB2 in the amygdala influences visceral sensitivity via glucocorticoid and mineralocorticoid receptors in male maternal separation rats [J]. Neurogastroenterol Motil, 2016, 28(10): 1545-1553.
[20] Mawe GM, Coates MD, Moses PL. Review article: Intestinal serotonin signalling in irritable bowel syndrome [J]. Aliment Pharmacol Ther, 2010, 23(8): 1067-1076.
[21] Gershon MD, Tack J. The serotonin signaling system: from basic understanding to drug development for functional GI disorders [J]. Gastroenterology, 2007, 132(1): 397-414.
[22] Bellono NW, Bayrer JR, Leitch DB, et al. Enterochromaffin cells are gut chemosensors that couple to sensory neural pathways [J]. Cell, 2017, 170(1): 185-198.
[2] Bradford K, Shih W, Videlock EJ, et al. Association between early adverse life events and irritable bowel syndrome [J].Clin Gastroenterol Hepatol, 2012, 10(4): 385-390.
[3] Van Oudenhove L, Crowell MD, Drossman DA, et al. Biopsychosocial aspects of functional gastrointestinal disorders :How central and environmental processes contribute to the development and expression of functional gastrointestinal disorders[J]. Gastroenterology, 2016,150(6):1355-1367.
[4] Tsang SW, Auyeung KK, Bian ZX, et al. Pathogenesis, experimental models and contemporary pharmacotherapy of irritable bowel syndrome: story about the brain-gut axis [J]. Curr Neuropharmacol, 2016,14(8): 842-856.
[5] Hungin AP, Becher A, Cayley B, et al. Irritable bowel syndrome: an integrated explanatory model for clinical practice [J]. Neurogastroenterol Motil, 2015, 27(6): 750-763.
[6] Keszthelyi D, Troost FJ, Masclee AA. Irritable bowel syndrome: methods, mechanisms, and pathophysiology. Methods to assess visceral hypersensitivity in irritable bowel syndrome [J]. Am J Physiol Gastrointest Liver Physiol, 2012, 303(2):G141-154.
[7] 耿婷婷,汪婷婷,姜劲峰,等,足底电击应激时程对大鼠焦虑样行为的影响[J]. 中国比较医学杂志,2016(01):69-72.Geng TT, Wang TT, Jiang JF, et al. The influence of footshock stress duration on anxiety-like behavior in rats [J]. Chin J Comp Med, 2016(01): 69-72.
[8] 汤球, 刘志学, 崔淑芳,等.大鼠抑郁症模型的建立与评价[J]. 实验动物科学, 2011, 28(1):6-9.Tao Q,Liu ZM,Cui SF,et al,Establishment and evaluation of rat model of depression [J]. Lab Anim Sci, 2011, 28(1):6-9.
[9] Bradesi S, Schwetz I, Ennes HS, et al. Repeated exposure to water avoidance stress in rats: a new model for sustained visceral hyperalgesia [J]. Am J Physiol Gastrointest Liver Physiol, 2005, 289(1): G42-53.
[10] Nozu T, Miyagishi S, Nozu R, et al. Water avoidance stress induces visceral hyposensitivity through peripheral corticotropin releasing factor receptor type 2 and central dopamine D2 receptor in rats [J]. Neurogastroenterol Motil, 2016, 28(4): 522-531.
[11] 张北华,王微,王凤云,等,基于不同精神心理应激建立IBS-D大鼠模型的比较研究[J]. 世界中医药,2017(09):2128-2133.Zhang BH, Wang W, Wang FY, et al. Comparative study of establishment of a rat model of IBS-D based on different psychological stress [J]. World Chin Med. 2017(09): 2128-2133.
[12] O’Mahony SM, Hyland NP, Dinan TG, et al. Maternal separation as a model of brain-gut axis dysfunction [J]. Psychopharmacology (Berl), 2011, 214(1): 71-88.
[13] Jia M, Lu X, Wang Z, et al. Effects of Fengliao-Changweikang in diarrhea-predominant irritable bowel syndrome rats and its mechanism involving colonic motility [J]. J Neurogastroenterol Motil, 2018, 24(3): 479-489.
[14] Kanada Y, Katayama A, Ikemoto H, et al. Inhibitory effect of the Kampo medicinal formula Yokukansan on acute stress-induced defecation in rats [J]. Neuropsychiatr Dis Treat, 2018, 14: 937-944.
[15] 刘晓萌,张荣,董世芬,等.乌灵菌粉贯叶连翘复方制剂干预对慢性不可预知应激大鼠模型抗抑郁作用及机制的研究[J]. 中国比较医学杂志,2016, 26(05):81-86.Liu XM,,Zhang R, Dong SF, et al. Effects of mixture of Wuling powder and Hypericum perforatum L. on depressed model rats induced by chronic unpredictable stress [J]. Chin J Comp Med, 2016, 26(05): 81-86.
[16] 郭维,孙琪,桂牧微,等.慢性应激损伤对消化系统相关血清肿瘤标志物的影响 [J]. 广东医学,2017(23):3558-3560.Guo W,Sun Q,Gui MW,et al. Experimental study on the effect of chronic stress damage on serum tumor markers [J].Guangdong Med J 2017(23): 3558-3560.
[17] Smith SM, Vale WW. The role of the hypothalamic-pituitary-adrenal axis in neuroendocrine responses to stress [J]. Dialogues Clin Neurosci, 2006, 8(4): 383-395.
[18] Nozu T, Okumura T. Corticotropin-releasing factor receptor type 1 and type 2 interaction in irritable bowel syndrome [J]. J Gastroenterol, 2015, 50(8): 819-830.
[19] Zhou XP, Sha J, Huang L, et al. Nesfatin-1/NUCB2 in the amygdala influences visceral sensitivity via glucocorticoid and mineralocorticoid receptors in male maternal separation rats [J]. Neurogastroenterol Motil, 2016, 28(10): 1545-1553.
[20] Mawe GM, Coates MD, Moses PL. Review article: Intestinal serotonin signalling in irritable bowel syndrome [J]. Aliment Pharmacol Ther, 2010, 23(8): 1067-1076.
[21] Gershon MD, Tack J. The serotonin signaling system: from basic understanding to drug development for functional GI disorders [J]. Gastroenterology, 2007, 132(1): 397-414.
[22] Bellono NW, Bayrer JR, Leitch DB, et al. Enterochromaffin cells are gut chemosensors that couple to sensory neural pathways [J]. Cell, 2017, 170(1): 185-198.