厚朴三物汤对术后肠梗阻大鼠小肠Cajal细胞及iNOS表达的影响
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摘要
目的:以Cajal间质细胞(ICC)及诱导型一氧化氮合酶(iNOS)表达为切入点,观察厚朴三物汤对术后肠梗阻大鼠小肠炎症反应与神经反馈的影响,并探讨其对胃肠道运动作用的相关机制。方法:健康成年雄性SD大鼠87只,随机分为假手术组、模型组、厚朴三物汤低、中、高剂量组。采用手术方法造模,造模后分别给予生理盐水或不同剂量的厚朴三物汤灌胃,低、中、高剂量组给药量分别为2. 25,4. 5,9 g·kg-1。给药后观察各组大鼠术后不同时间的小肠推进率及小肠形态学变化,免疫组织化学法检测ICC细胞及iNOS的表达情况。结果:与假手术组比较,造模后大鼠小肠推进率明显下降(P <0. 01),造模后可见小肠肠管明显扩张、胀气,苏木素-伊红(HE)染色可见小肠绒毛破裂,结构不完整,黏膜下可见血管充血水肿。与盐水组比较,厚朴三物汤中、高剂量组大鼠24,48 h小肠推进率显著提高(P <0. 01),而且厚朴三物汤治疗后3组在48 h内大鼠小肠推进率均随时间延长而增加(P <0. 01)。HE染色可见大鼠肠黏膜绒毛脱落及组织水肿程度较盐水组明显减轻。免疫组化显示造模后大鼠小肠Cajal间质细胞表达下降,iNOS阳性表达显著升高;而厚朴三物汤治疗后3组Cajal间质细胞在24,48 h均显著升高,中、低剂量组12,24 h iNOS阳性表达明显降低(P <0. 05,P <0. 01)。结论:厚朴三物汤能够显著提高术后肠梗阻模型大鼠的小肠推进率,促进大鼠术后胃肠动力的恢复,其作用机制与抑制iNOS的产生,减轻炎症反应,同时增加ICC数量,保证其正常功能,从而改善肠动力障碍有关。
Objective: To evaluate the effect of Houpu Sanwu Tang on the postoperative ileus(POI),and observe its underlying mechanisms of action on interstitial cells of cajal(ICC) and inducible nitric oxide synthase(iNOS) regulation of POI. Method: Totally 87 healthy adult male SD rats were randomly divided into sham operation group,saline control group and Houpu Sanwu Tang group at low,medium and high doses. Houpu Sanwu Tang low,middle and high dose groups received orally Houpu Sanwu Tang(2. 25,4. 5,9 g · kg-1);Sham operation group(Sham operation) and saline control group(Saline control) received orally normal saline.Surgical procedure was used to induce the postoperative ileus. Changes in intestinal propulsion rate,intestinal mucosal injury and small intestine expression of c-kit and iNOS among these groups were detected. Result:Intestinal propulsion rate was significantly higher in Houpo Sanwu Tang group than that in Saline control group(P < 0. 01). Rats in Houpo Sanwu Tang group showed reduced intestinal mucosal injury compared with those in Saline control group. C-kit in Houpo Sanwu Tang groups was significantly increased(P < 0. 01) at 24 h and 48 h,and iNOS in low-dose group and middle-dose group was significantly reduced(P < 0. 05,P < 0. 01) at 12 h and24 h compared with those in the Saline control group. Conclusion: Houpu Sanwu Tang can improve the intestinal propulsion rate and the recovery in POI rats. The mechanisms are related to the inhibition of the generation of iNOS,the alleviation of inflammatory response,and increase of the number of ICC,so as to ensure its normal function,and improve the intestinal dynamic disorder.
Objective: To evaluate the effect of Houpu Sanwu Tang on the postoperative ileus(POI),and observe its underlying mechanisms of action on interstitial cells of cajal(ICC) and inducible nitric oxide synthase(iNOS) regulation of POI. Method: Totally 87 healthy adult male SD rats were randomly divided into sham operation group,saline control group and Houpu Sanwu Tang group at low,medium and high doses. Houpu Sanwu Tang low,middle and high dose groups received orally Houpu Sanwu Tang(2. 25,4. 5,9 g · kg-1);Sham operation group(Sham operation) and saline control group(Saline control) received orally normal saline.Surgical procedure was used to induce the postoperative ileus. Changes in intestinal propulsion rate,intestinal mucosal injury and small intestine expression of c-kit and iNOS among these groups were detected. Result:Intestinal propulsion rate was significantly higher in Houpo Sanwu Tang group than that in Saline control group(P < 0. 01). Rats in Houpo Sanwu Tang group showed reduced intestinal mucosal injury compared with those in Saline control group. C-kit in Houpo Sanwu Tang groups was significantly increased(P < 0. 01) at 24 h and 48 h,and iNOS in low-dose group and middle-dose group was significantly reduced(P < 0. 05,P < 0. 01) at 12 h and24 h compared with those in the Saline control group. Conclusion: Houpu Sanwu Tang can improve the intestinal propulsion rate and the recovery in POI rats. The mechanisms are related to the inhibition of the generation of iNOS,the alleviation of inflammatory response,and increase of the number of ICC,so as to ensure its normal function,and improve the intestinal dynamic disorder.
引文
[1]袁凯涛,石汉平.术后肠梗阻的病因与预防[J].中华普通外科学文献:电子版,2010,4(6):588-591.
[2] Carroll J,Alavi K. Pathogenesis and management of postoperative ileus[J]. Clin Colon Rectal Surg,2009,22(1):47-50.
[3] Boeckxstaens G E, de Jonge W J. Neuroimmune mechanismsin postoperative ileus[J]. Gut,2009,58(9):1300-1311.
[4]刘桂芳,耿涛,房玉涛.糖胃安煎剂对糖尿病胃肠病变大鼠Cajal间质细胞和P物质的影响[J].中国实验方剂学杂志,2018,24(22):127-132.
[5]周晴晴,曹立幸,陈志强.中医药对围手术期术后胃肠功能恢复作用的研究进展[J].中国中西医结合杂志,2013,33(6):858-860.
[6]范永升,姜德友.金匮要略[M]. 3版.北京:中国中医药出版社,2014:122.
[7]李慎贤,王宝胜,王廷振.厚朴三物汤空肠给药促进胃十二指肠术后胃肠功能恢复的临床观察[J].中国中西医结合急救杂志,2005,5,12(3):162-164.
[8]张晓东,顾群浩,蔡照弟,等.厚朴三物汤治疗不完全性肠梗阻52例临床观察[J].河北中医,2012,34(6):852-853.
[9]王殿民.全成分中药配方颗粒葛根汤治疗颈椎病的疗效观察[J].西部医学,2011,23(8):1531-1533.
[10] Kalff J C,Buchholz B M,Eskandari M K,et al.Biphasic response to gut manipulation and temporal correlation of cellular infiltrates and muscle dysfunction in rat[J]. Surgery,1999,126(3):498-509.
[11]陈奇.中药药理学实验方法学[M].北京:人民卫生出版社,1994:55.
[12] Remmele W,Stegner H E. Recommendation for uniform definition of an immunoreactive score(IRS)for immunohistochemical estrogen receptor detection(ERICA)in breast cancer tissue[J]. Pathology,1987,8(3):138-140.
[13]吴咸中.腹部外科实践[M]. 3版.天津:天津科学技术出版社,2004:866-869.
[14] Kalff J C,Schraut W H,Simmons R L,et al. Surgical manipulation of the gut elicits an intestinal muscularis inflammatory response resulting in postsurgical ileus[J]. Ann Surg,1998,228(5):652-663.
[15] Ohama T,Hori M,Ozaki H. Mechanism of abnormal intestinal motility in inflammatory bowel disease:how smooth muscle contraction is reduced[J]. Smooth Muscle Res,2007,43(2):43-54.
[16]李小玲,林中,袁园,等.胃肠动力的神经调节[J].神经解剖学杂志,2012,28(2):213-216.
[17]周英豪,吉敏,李成林. Cajal间质细胞与胃肠道疾病[J].医学综述,2012,18(13):1998-2001.
[18] LI J,Goodyer C G,Fellows F,et al. Stem cell factor/c-Kit interactions regulate human islet-epithelial cluster proliferation and differentiation[J]. Int J Biochem Cell Biol,2006,38(5/6):961-972.
[19] Bashamboo A,Taylor A H,Samuel K,et al. The survival of differentiating embryonic stem cells is dependent on the SCF-KIT pathway[J]. J Cell Sci,2006,119(15):3039-3046.
[20]蒋峰,陈玉根.维持Cajal间质细胞网络机制的研究进展[J].江苏医药,2017,43(18):1338-1341.
[21] Vannucchi M G. Receptors in interstitial cells of Cajal:identification and possible physiological roles[J].Microsc Res Tech,1999,47(5):325-335.
[22]田姣.胃肠道Cajal间质细胞与干细胞因子/c-kit信号系统的研究进展[J].临床儿科杂志,2013,31(4):385-388.
[23] Derbyshire E R,Marletta M A. Structure and regulation of soluble guanylate cyclase[J]. Annu Rev Biochem,2012,81:533-539.
[24] Iino S,Horiguchi K,Nojyo Y,et al. Interstitial cells of Cajal contain signalling molecules for transduction of nitrergic stimulation in guinea pig caecum[J].Neurogastroenterol Motil,2009,21(5):542-550.
[25]瞿金鸿,李炯侠,王子坪.“六腑以通为用”在临床中的应用[J].辽宁中医药大学学报,2008,10(6):21-22.
[26]高聪,韩勇,林大勇,等.加味桃核承气汤内服和灌肠对腹部术后早期肠梗阻神经-炎症机制的影响[J].中国实验方剂学杂志,2017,23(16):176-181.
[27] Kalff J C,Buchholz B M,Eskandari M K,el at.Biphasic response to gut manipulation and temporal correlation of cellular infiltrates and muscle dysfunction in rat[J]. Surgery,1999,126(3):498-509.
[28]王立茹.鼻咽合剂不同剂型治疗慢性鼻炎咽炎疗效比对研究[J].陕西中医,2012,33(6):702-703.
[29]余轶群,陈飞松.健脾消满中药配方颗粒治疗痞满证的临床观察[J].北京中医,2007,26(4):229-231.
[2] Carroll J,Alavi K. Pathogenesis and management of postoperative ileus[J]. Clin Colon Rectal Surg,2009,22(1):47-50.
[3] Boeckxstaens G E, de Jonge W J. Neuroimmune mechanismsin postoperative ileus[J]. Gut,2009,58(9):1300-1311.
[4]刘桂芳,耿涛,房玉涛.糖胃安煎剂对糖尿病胃肠病变大鼠Cajal间质细胞和P物质的影响[J].中国实验方剂学杂志,2018,24(22):127-132.
[5]周晴晴,曹立幸,陈志强.中医药对围手术期术后胃肠功能恢复作用的研究进展[J].中国中西医结合杂志,2013,33(6):858-860.
[6]范永升,姜德友.金匮要略[M]. 3版.北京:中国中医药出版社,2014:122.
[7]李慎贤,王宝胜,王廷振.厚朴三物汤空肠给药促进胃十二指肠术后胃肠功能恢复的临床观察[J].中国中西医结合急救杂志,2005,5,12(3):162-164.
[8]张晓东,顾群浩,蔡照弟,等.厚朴三物汤治疗不完全性肠梗阻52例临床观察[J].河北中医,2012,34(6):852-853.
[9]王殿民.全成分中药配方颗粒葛根汤治疗颈椎病的疗效观察[J].西部医学,2011,23(8):1531-1533.
[10] Kalff J C,Buchholz B M,Eskandari M K,et al.Biphasic response to gut manipulation and temporal correlation of cellular infiltrates and muscle dysfunction in rat[J]. Surgery,1999,126(3):498-509.
[11]陈奇.中药药理学实验方法学[M].北京:人民卫生出版社,1994:55.
[12] Remmele W,Stegner H E. Recommendation for uniform definition of an immunoreactive score(IRS)for immunohistochemical estrogen receptor detection(ERICA)in breast cancer tissue[J]. Pathology,1987,8(3):138-140.
[13]吴咸中.腹部外科实践[M]. 3版.天津:天津科学技术出版社,2004:866-869.
[14] Kalff J C,Schraut W H,Simmons R L,et al. Surgical manipulation of the gut elicits an intestinal muscularis inflammatory response resulting in postsurgical ileus[J]. Ann Surg,1998,228(5):652-663.
[15] Ohama T,Hori M,Ozaki H. Mechanism of abnormal intestinal motility in inflammatory bowel disease:how smooth muscle contraction is reduced[J]. Smooth Muscle Res,2007,43(2):43-54.
[16]李小玲,林中,袁园,等.胃肠动力的神经调节[J].神经解剖学杂志,2012,28(2):213-216.
[17]周英豪,吉敏,李成林. Cajal间质细胞与胃肠道疾病[J].医学综述,2012,18(13):1998-2001.
[18] LI J,Goodyer C G,Fellows F,et al. Stem cell factor/c-Kit interactions regulate human islet-epithelial cluster proliferation and differentiation[J]. Int J Biochem Cell Biol,2006,38(5/6):961-972.
[19] Bashamboo A,Taylor A H,Samuel K,et al. The survival of differentiating embryonic stem cells is dependent on the SCF-KIT pathway[J]. J Cell Sci,2006,119(15):3039-3046.
[20]蒋峰,陈玉根.维持Cajal间质细胞网络机制的研究进展[J].江苏医药,2017,43(18):1338-1341.
[21] Vannucchi M G. Receptors in interstitial cells of Cajal:identification and possible physiological roles[J].Microsc Res Tech,1999,47(5):325-335.
[22]田姣.胃肠道Cajal间质细胞与干细胞因子/c-kit信号系统的研究进展[J].临床儿科杂志,2013,31(4):385-388.
[23] Derbyshire E R,Marletta M A. Structure and regulation of soluble guanylate cyclase[J]. Annu Rev Biochem,2012,81:533-539.
[24] Iino S,Horiguchi K,Nojyo Y,et al. Interstitial cells of Cajal contain signalling molecules for transduction of nitrergic stimulation in guinea pig caecum[J].Neurogastroenterol Motil,2009,21(5):542-550.
[25]瞿金鸿,李炯侠,王子坪.“六腑以通为用”在临床中的应用[J].辽宁中医药大学学报,2008,10(6):21-22.
[26]高聪,韩勇,林大勇,等.加味桃核承气汤内服和灌肠对腹部术后早期肠梗阻神经-炎症机制的影响[J].中国实验方剂学杂志,2017,23(16):176-181.
[27] Kalff J C,Buchholz B M,Eskandari M K,el at.Biphasic response to gut manipulation and temporal correlation of cellular infiltrates and muscle dysfunction in rat[J]. Surgery,1999,126(3):498-509.
[28]王立茹.鼻咽合剂不同剂型治疗慢性鼻炎咽炎疗效比对研究[J].陕西中医,2012,33(6):702-703.
[29]余轶群,陈飞松.健脾消满中药配方颗粒治疗痞满证的临床观察[J].北京中医,2007,26(4):229-231.