电针不同穴组对功能性腹泻大鼠下丘脑及结肠5-羟色胺和c-fos蛋白表达的影响
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摘要
目的:通过观察电针不同组穴对功能性腹泻(FD)大鼠下丘脑及结肠组织5-羟色胺1A受体(5-HT_(1A)R)、c-fos蛋白表达的影响,初步探讨电针对FD大鼠脑-肠关系的影响。方法:雄性SD大鼠随机分为空白组、模型组、电针Ⅰ组、电针Ⅱ组,每组10只。采用番泻叶加束缚制动的方法建立FD大鼠模型。电针Ⅰ组电针大鼠双侧"天枢""大肠俞",电针Ⅱ组电针大鼠双侧"曲池""上巨虚",均留针20min,每日1次,连续10d。观察并记录各组大鼠治疗前后稀便率及腹泻指数。Western blot法检测各组大鼠下丘脑及结肠组织5-HT_(1A)R、c-fos蛋白表达水平。结果:治疗前,与空白组比较,模型组、电针Ⅰ组、电针Ⅱ组稀便率及腹泻指数均升高(P<0.01)。治疗后,与空白组比较,模型组稀便率及腹泻指数均升高(P<0.01);与模型组比较,电针Ⅰ组稀便率及腹泻指数均降低(P<0.01);与电针Ⅰ组比较,电针Ⅱ组稀便率及腹泻指数显著升高(P<0.01)。与空白组比较,模型组下丘脑及结肠组织中5-HT_(1A)R和c-fos蛋白表达水平均升高(P<0.05);与模型组比较,电针Ⅰ组下丘脑及结肠组织中5-HT_(1A)R及c-fos蛋白表达水平降低(P<0.05),电针Ⅱ组结肠组织中5-HT_(1A)R蛋白表达水平降低(P<0.05);与电针Ⅰ组比较,电针Ⅱ组大鼠下丘脑中5-HT_(1A)R、下丘脑及结肠组织中c-fos蛋白表达水平均升高(P<0.05)。结论:电针"天枢""大肠俞"可减轻番泻叶诱导的FD大鼠的腹泻症状,降低下丘脑及结肠组织5-HT_(1A)R和c-fos的表达,其机制可能与脑-肠轴系统有关。
Objective To observe the effect of electroacupuncture(EA)on expression of 5-HT_(1A)receptor(5-HT_(1A)R and c-fos proteins in the hypothalamus and colon tissues in functional diarrhea(FD)rats so as to explore its underlying mechanisms in improving intestinal function via brain-gut axis.Methods A total of 40 SD rats were randomized into blank control,model,EA Tianshu(ST25)-Dachangshu(BL25,ST25-BL25)and EA Quchi(LI11)-Shangjuxu(ST37,LI11-ST37)groups.The FD model was established by gavage of Folium Sennae(10 mg/kg)and constraint immobilization once daily for 29 days.EA(10 Hz/50 Hz,1.5 mA)was applied to bilateral ST25 and BL25 in EA ST25-BL25 group,and bilateral LI11 and ST37 in EA LI11-ST37 group for20 min,once daily for successive 10 days.The expression of 5-HT_(1A)R and c-fos protein in the hypothalamus and colon tissues was determined by using Western blot.The state of stool was recorded for calculating the loose stool rate and diarrhea index.Results After modeling,the loose stool rate,diarrhea index and expression levels of 5-HT_(1A)R and c-fos proteins in the colon and hypothalamus tissues were obviously increased in the model group in contrast with the blank control group(P<0.01,P<0.05).Following EA intervention,the loose stool rate and diarrhea index,the expression levels of 5-HT_(1A)R and c-fos proteins in the hypothalamus and colon in the EA ST25-BL25 group,and the expression of 5-HT_(1A)R in the colon in the EA LI11-ST37 group were significantly downregulated relevant to the model group(P<0.01,P<0.05).No significant changes were found in loose stool rate and diarrhea index,and the expression levels of hypothalamic 5-HT_(1A)R and hypothalamic and colonic c-fos proteins in the EA LI11-ST37 group(P>0.05).The expression levels of 5-HT_(1A)R protein in the hypothalamus and c-fos protein in the hypothalamus and colon in the EA LI11-ST37 group were significantly up-regulated relevant to the EA ST25-BL25 group(P<0.05).Conclusion EA of ST25-BL25 can down-regulate expression of 5-HT_(1A)R and c-fos proteins in the hypothalamus and colon tissue in FD rats,which may contribute to its function in improving symptoms of FD possibly via brain-gut axis.
Objective To observe the effect of electroacupuncture(EA)on expression of 5-HT_(1A)receptor(5-HT_(1A)R and c-fos proteins in the hypothalamus and colon tissues in functional diarrhea(FD)rats so as to explore its underlying mechanisms in improving intestinal function via brain-gut axis.Methods A total of 40 SD rats were randomized into blank control,model,EA Tianshu(ST25)-Dachangshu(BL25,ST25-BL25)and EA Quchi(LI11)-Shangjuxu(ST37,LI11-ST37)groups.The FD model was established by gavage of Folium Sennae(10 mg/kg)and constraint immobilization once daily for 29 days.EA(10 Hz/50 Hz,1.5 mA)was applied to bilateral ST25 and BL25 in EA ST25-BL25 group,and bilateral LI11 and ST37 in EA LI11-ST37 group for20 min,once daily for successive 10 days.The expression of 5-HT_(1A)R and c-fos protein in the hypothalamus and colon tissues was determined by using Western blot.The state of stool was recorded for calculating the loose stool rate and diarrhea index.Results After modeling,the loose stool rate,diarrhea index and expression levels of 5-HT_(1A)R and c-fos proteins in the colon and hypothalamus tissues were obviously increased in the model group in contrast with the blank control group(P<0.01,P<0.05).Following EA intervention,the loose stool rate and diarrhea index,the expression levels of 5-HT_(1A)R and c-fos proteins in the hypothalamus and colon in the EA ST25-BL25 group,and the expression of 5-HT_(1A)R in the colon in the EA LI11-ST37 group were significantly downregulated relevant to the model group(P<0.01,P<0.05).No significant changes were found in loose stool rate and diarrhea index,and the expression levels of hypothalamic 5-HT_(1A)R and hypothalamic and colonic c-fos proteins in the EA LI11-ST37 group(P>0.05).The expression levels of 5-HT_(1A)R protein in the hypothalamus and c-fos protein in the hypothalamus and colon in the EA LI11-ST37 group were significantly up-regulated relevant to the EA ST25-BL25 group(P<0.05).Conclusion EA of ST25-BL25 can down-regulate expression of 5-HT_(1A)R and c-fos proteins in the hypothalamus and colon tissue in FD rats,which may contribute to its function in improving symptoms of FD possibly via brain-gut axis.
引文
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[3]TALLEY N J,HOWELL S,POULTON R.The irritable bowel syndrome and psychiatric disorders in the community:is there alink[J].Am J Gastroenterol,2001,96(4):1072-1079.
[4]李兆申,詹丽杏,邹多武,等.肠易激综合征患者分泌5-羟色胺的肠嗜铬细胞形态及功能的改变[J].中华消化杂志,2004,24(2):94-97.
[5]DARVISH-DAMAVADI M,NIKFAR S,ABDOLLAHI M.A systermatic reviw of efficacy and tolerability of mebeverine in irritable bowel syndrome[J].Word J Gastroenterol,2010,16(5):547-553.
[6]AIT-BELGNAOUI A,EUTAMENE H,HOUDEAU E,et al.Lactobacillus farciminis treatment attenuates stress-induced overexpression of Fos protein in spinal and supraspinal sites after colorectal distension in rats[J].Neurogastroenterol Motil,2009,21(5):567-573.
[7]VAZQUEZ-ROQUE M I,CAMILLERI M,CARLSON P,et al.HLA-DQ genotype is associated with accelerated small bowel transit in patients with diarrhea-predominant irritable bowel syndrome[J].Eur J Gastroenterol Hepatol,2011,23(6):481-487.
[8]AWAD R A.Neurogenic bowel dysfunction in patients with spinal cord injury,myelomeningocele,multiple sclerosis and Parkinsons disease[J].World J Gastroenterol,2011,17(46):5035-5048.
[9]梁莉婕,焦立波,高明.建立动物腹泻模型的常用药物及方法的研究进展[J].辽宁中医杂志,2011,38(8):1687-1689.
[10]包新民,舒斯云.大鼠脑立体定位图谱[M].北京:人民卫生出版社,1991.
[11]BOSE M,FARTHING M J.Irritable bowel syndrome:new horizons in pathophysiology and treatment[J].Br J Surg,2001,88(11):1425-1426.
[12]COELHO A M,JACOB L,FIORAMONTI J,et al.Rectal antinociceptive properties of alverine citrate are linked to antagonism at the 5-HT1A receptor subtype[J].J Pharm Pharmacol,2001,53(10):1419-1426.
[13]SUZUKI R,RYGH L J,DICKENSON A H.Bad news from the brain:descending 5-HT pathways that control spinal pain processing[J].Trends Pharmacol Sci,2004,25(12):613-617.
[14]CAMILLERI M,MAYER E A,DROSSMAN D A,et al.Improvement in pain and bowel function in female irritable bowel patients with alosetron,a 5-HT3receptor antagonist[J].Aliment Pharmacol Ther,1999,13(9):1149-1159.
[15]GORARD D A,DEWSNAP P A,MEDBAK S H,et al.Central 5-hydroxytryptaminergic function in irritable bowel syndrome[J].Scand J Gastroenterol,1995,30(10):994-999.
[16]孙刚,杨云生,彭丽华,等.肠易激综合征大鼠内脏敏感性异常与结肠及中枢神经系统5-HT和C-fos表达的关系[J].胃肠病学和肝病学杂志,2008,17(4):313-317.
[17]ZIGHELBOIM J,TALLEY N J,PHILLIPS S F,et al.Visceralperception in irritable bowel syndrome.rectal and gastric responses to distension and serotonin type 3antagonism[J].Dig Dis Sci,1995,40(4):819-827.
[18]卢晓虹,李凌江,李昌琦,等.应激对中枢神经系统即刻早期基因c-fos表达及HPA轴的调节作用研究[J].中国心理卫生杂志,2000,14(1):10-13.
[19]RINGEL Y,DROSSMAN D A,TURKINGTON T G,et al.Regional brain activation in response to rectal distension in patients with irritable bowel syndrome and the effect of a history of abuse[J].Dig Dis Sci,2003,48(9):1774-1781.
[20]胡品津.从脑-肠互动的高度认识肠易激综合征[J].中华消化杂志,2003,23(5):261-262.
[21]GAO J,WU X Y,OWYANG C,et al.Enhanced responses of the anteriorcingulate cortex neurones to colonic distension in viscerally hypersensitive rats[J].J Physiol(Lond),2006,570(Pt1):169-183.
[22]王渊,刘智斌,牛文民,等.电针不同穴位对功能性腹泻大鼠血清和组织中P物质、血管活性肠肽的影响[J].广州中医药大学学报,2012,29(6):656-659.
[23]朱兵.针灸双向调节效应的生物学意义[J].世界中医药,2013,8(3):241-244.
[2]EMMANUEL A V,MASON H J,KAMM M A.Relationship between psychologicalstate and level of activity of extrinsic gut innervation in patients with a functional gut disorder[J].Gut,2001,49(2):209-213.
[3]TALLEY N J,HOWELL S,POULTON R.The irritable bowel syndrome and psychiatric disorders in the community:is there alink[J].Am J Gastroenterol,2001,96(4):1072-1079.
[4]李兆申,詹丽杏,邹多武,等.肠易激综合征患者分泌5-羟色胺的肠嗜铬细胞形态及功能的改变[J].中华消化杂志,2004,24(2):94-97.
[5]DARVISH-DAMAVADI M,NIKFAR S,ABDOLLAHI M.A systermatic reviw of efficacy and tolerability of mebeverine in irritable bowel syndrome[J].Word J Gastroenterol,2010,16(5):547-553.
[6]AIT-BELGNAOUI A,EUTAMENE H,HOUDEAU E,et al.Lactobacillus farciminis treatment attenuates stress-induced overexpression of Fos protein in spinal and supraspinal sites after colorectal distension in rats[J].Neurogastroenterol Motil,2009,21(5):567-573.
[7]VAZQUEZ-ROQUE M I,CAMILLERI M,CARLSON P,et al.HLA-DQ genotype is associated with accelerated small bowel transit in patients with diarrhea-predominant irritable bowel syndrome[J].Eur J Gastroenterol Hepatol,2011,23(6):481-487.
[8]AWAD R A.Neurogenic bowel dysfunction in patients with spinal cord injury,myelomeningocele,multiple sclerosis and Parkinsons disease[J].World J Gastroenterol,2011,17(46):5035-5048.
[9]梁莉婕,焦立波,高明.建立动物腹泻模型的常用药物及方法的研究进展[J].辽宁中医杂志,2011,38(8):1687-1689.
[10]包新民,舒斯云.大鼠脑立体定位图谱[M].北京:人民卫生出版社,1991.
[11]BOSE M,FARTHING M J.Irritable bowel syndrome:new horizons in pathophysiology and treatment[J].Br J Surg,2001,88(11):1425-1426.
[12]COELHO A M,JACOB L,FIORAMONTI J,et al.Rectal antinociceptive properties of alverine citrate are linked to antagonism at the 5-HT1A receptor subtype[J].J Pharm Pharmacol,2001,53(10):1419-1426.
[13]SUZUKI R,RYGH L J,DICKENSON A H.Bad news from the brain:descending 5-HT pathways that control spinal pain processing[J].Trends Pharmacol Sci,2004,25(12):613-617.
[14]CAMILLERI M,MAYER E A,DROSSMAN D A,et al.Improvement in pain and bowel function in female irritable bowel patients with alosetron,a 5-HT3receptor antagonist[J].Aliment Pharmacol Ther,1999,13(9):1149-1159.
[15]GORARD D A,DEWSNAP P A,MEDBAK S H,et al.Central 5-hydroxytryptaminergic function in irritable bowel syndrome[J].Scand J Gastroenterol,1995,30(10):994-999.
[16]孙刚,杨云生,彭丽华,等.肠易激综合征大鼠内脏敏感性异常与结肠及中枢神经系统5-HT和C-fos表达的关系[J].胃肠病学和肝病学杂志,2008,17(4):313-317.
[17]ZIGHELBOIM J,TALLEY N J,PHILLIPS S F,et al.Visceralperception in irritable bowel syndrome.rectal and gastric responses to distension and serotonin type 3antagonism[J].Dig Dis Sci,1995,40(4):819-827.
[18]卢晓虹,李凌江,李昌琦,等.应激对中枢神经系统即刻早期基因c-fos表达及HPA轴的调节作用研究[J].中国心理卫生杂志,2000,14(1):10-13.
[19]RINGEL Y,DROSSMAN D A,TURKINGTON T G,et al.Regional brain activation in response to rectal distension in patients with irritable bowel syndrome and the effect of a history of abuse[J].Dig Dis Sci,2003,48(9):1774-1781.
[20]胡品津.从脑-肠互动的高度认识肠易激综合征[J].中华消化杂志,2003,23(5):261-262.
[21]GAO J,WU X Y,OWYANG C,et al.Enhanced responses of the anteriorcingulate cortex neurones to colonic distension in viscerally hypersensitive rats[J].J Physiol(Lond),2006,570(Pt1):169-183.
[22]王渊,刘智斌,牛文民,等.电针不同穴位对功能性腹泻大鼠血清和组织中P物质、血管活性肠肽的影响[J].广州中医药大学学报,2012,29(6):656-659.
[23]朱兵.针灸双向调节效应的生物学意义[J].世界中医药,2013,8(3):241-244.
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