不同灸量隔药灸对溃疡性结肠炎大鼠结肠组织病理学的影响
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摘要
目的:观察隔药灸不同灸量对溃疡性结肠炎(UC)大鼠结肠损伤的作用差异,从组织病理学角度筛选隔药灸干预UC大鼠的最佳灸量组合。方法:SD大鼠随机分为正常组、模型组、西药组、隔药灸1组、隔药灸2组、隔药灸3组、隔药灸4组,每组9只。采用抗原免疫加局部化学刺激法制备UC模型。隔药灸各组选取双侧"天枢"穴给予不同灸量组合(每日1壮、每日2壮、隔日1壮、隔日2壮)隔药灸治疗。观察大鼠一般情况,苏木素-伊红(HE)染色观察结肠组织形态变化,过碘酸雪夫(PAS)染色观察结肠黏蛋白分泌情况,高铁二胺-阿辛蓝(HID-AB)染色观察结肠黏蛋白硫酸化程度。结果:与正常组比较,模型组大鼠便稀,肛周脓血分泌,体质量增加缓慢(P<0.01);巨检显示模型大鼠结肠充血明显,严重者局部存在数个浅小溃疡;HE染色镜检显示结肠浅表性溃疡连续分布,腺腔不规则,黏膜固有层、黏膜下层水肿伴大量炎性细胞浸润,组织损伤评分显著升高(P<0.01);PAS染色、HID-AB染色显示结肠肠腺黏蛋白明显减少(P<0.01)。与模型组比较,西药组及隔药灸各组大鼠均体质量增加(P<0.01,P<0.05),大便成形,结肠组织损伤改善,表现为黏膜愈合和炎性反应减轻,损伤评分明显降低(P<0.01);PAS染色、HID-AB染色显示隔药灸各组大鼠结肠腺体分泌黏蛋白增加(P<0.01)。两因素两水平析因分析结果显示,隔日灸对组织学评分的改善作用优于每日灸(P<0.05),隔日灸2壮对黏蛋白硫酸化程度的调节效应优于1壮(P<0.01)。隔药灸壮数与频次对结肠组织评分、黏蛋白分泌均有交互作用,共同影响灸效。结论:隔药灸"天枢"穴具有促进UC大鼠结肠损伤修复及黏液分泌作用,隔日2壮灸为最佳灸量组合。
Objective To observe the effect of different-doses of herbal cake-partitioned moxibustion(Moxi)on histopathological changes of the damaged colon tissue in rats with ulcerative colitis(UC),so as to select an optimal dosage of Moxi in the treatment of UC.Methods Sixty-three male Sprague Dawley(SD)rats were randomized into 7 groups:normal control,model,medication,and 1,2,3 and 4 moxa-cone Moxi(n=9 rats per group).The UC model was developed by subcutaneous injection of emulsion(1 mL)containing colon mucosa-prepared protein suspension and complete Freund's adjuvant into the toes,groin and back.On the 38 th day,enema of 3%formalin and the aforementioned emulsion was used.Herbal-cake(composed of monkshood,cinnamon,etc.)partitioned Moxi with 1 or 2 moxa-cones(about 5 min/cone)was applied to bilateral"Tianshu"(ST 25)once daily or once every other day.The rat's general conditions(diet,movement,response ability,stool,and body weight)were observed,and histopathological changes(adhesion,ulcer formation and inflammation)of colon tissues were examined after hematoxylin-eosin(HE)staining,and scored(histopathological score).Gross score was given according to the severity of adhesion,ulcer formation and inflammation of colonic tissues under stereo microscope.The average optical density(AOD)values of colonic mucins were detected after periodic acid-schiff(PAS)staining,and those of the sulfated mucus content detected after high iron diamine-alcian blue(HID-AB)staining.Results Compared with the normal group,rats in the model group presented loose stool,or with pus and blood,and slowly increased body weight(P<0.01),obvious congestion of colon with ulcer spots or continuous superficial ulcer regions which had irregular glandular cavities,swelling and serious inflammatory infiltration in lamina propria and submucosa,and increased score of colon tissue damage(P<0.01).PAS and HID-AB staining showed a marked decrease of AOD values of colonic mucins and sulfated mucus in the model group relevant to the normal control group(P<0.01),suggesting a reduction of mucus secretion of intestinal glands.Following the intervention,rats in the Moxi groups presented an increase of the body weight,formed feces,and an improvement of the damaged colon tissues as mucosal healing and inflammatory reduction,and a marked decrease of the damage score relevant to the model rats.No significant differences were found in the gross scores among the medication,1,2,3 and 4 moxa-cone groups(P>0.05).The histopathological scores were significantly lower in the1 and 4 moxa-cone Moxi groups than in the medication group(P<0.05,P<0.01);and significantly lower in the 1,3,4 moxacone Moxi groups than in the 2 moxa-cone Moxi group(P<0.05,P<0.01).PAS staining showed a significant increase of the AOD values of colonic mucins in the 1,2,3 and 4 moxa-cone and medication groups relevant to the model group(P<0.01);and the AOD values of colonic mucins in the 1,3,4 moxa-cone Moxi groups were significantly increased than that in the 2 moxa-cone Moxi group(P<0.05,P<0.01).HID-AB staining showed that the AOD values of sulfated mucus content were significantly higher in the 2 and 4 moxa-cone Moxi groups than in the 3 moxa-cone Moxi group(P<0.01).The two-level two-factor factorial analysis showed an interaction existed between the moxa-cone number and Moxi frequency in reducing the gross score and histopathological score and in facilitating colonic mucin and sulfated mucus secretion.The histopathological score of the 4 moxa-cone Moxi group was significantly lower than that of the 2 moxa-cone Moxi group(P<0.05),and the sulfated mucus content was significantly higher in the 4 moxa-cone group than in the 3 moxa-cone group(P<0.01).The effect of Moxi given on alternate days was superior to that of daily Moxi in improving colonic histological damage.Conclusion Herbal cake-partitioned moxibustion at ST 25 can promote repair of the damaged colonic tissue and secretion of mucin in UC rats.The number of moxa cones and intervention frequency affect the efficacy of Moxi in improving histopathological changes.The Moxi intervention on alternate days and with 2 moxa-cones every time is recommended.
Objective To observe the effect of different-doses of herbal cake-partitioned moxibustion(Moxi)on histopathological changes of the damaged colon tissue in rats with ulcerative colitis(UC),so as to select an optimal dosage of Moxi in the treatment of UC.Methods Sixty-three male Sprague Dawley(SD)rats were randomized into 7 groups:normal control,model,medication,and 1,2,3 and 4 moxa-cone Moxi(n=9 rats per group).The UC model was developed by subcutaneous injection of emulsion(1 mL)containing colon mucosa-prepared protein suspension and complete Freund's adjuvant into the toes,groin and back.On the 38 th day,enema of 3%formalin and the aforementioned emulsion was used.Herbal-cake(composed of monkshood,cinnamon,etc.)partitioned Moxi with 1 or 2 moxa-cones(about 5 min/cone)was applied to bilateral"Tianshu"(ST 25)once daily or once every other day.The rat's general conditions(diet,movement,response ability,stool,and body weight)were observed,and histopathological changes(adhesion,ulcer formation and inflammation)of colon tissues were examined after hematoxylin-eosin(HE)staining,and scored(histopathological score).Gross score was given according to the severity of adhesion,ulcer formation and inflammation of colonic tissues under stereo microscope.The average optical density(AOD)values of colonic mucins were detected after periodic acid-schiff(PAS)staining,and those of the sulfated mucus content detected after high iron diamine-alcian blue(HID-AB)staining.Results Compared with the normal group,rats in the model group presented loose stool,or with pus and blood,and slowly increased body weight(P<0.01),obvious congestion of colon with ulcer spots or continuous superficial ulcer regions which had irregular glandular cavities,swelling and serious inflammatory infiltration in lamina propria and submucosa,and increased score of colon tissue damage(P<0.01).PAS and HID-AB staining showed a marked decrease of AOD values of colonic mucins and sulfated mucus in the model group relevant to the normal control group(P<0.01),suggesting a reduction of mucus secretion of intestinal glands.Following the intervention,rats in the Moxi groups presented an increase of the body weight,formed feces,and an improvement of the damaged colon tissues as mucosal healing and inflammatory reduction,and a marked decrease of the damage score relevant to the model rats.No significant differences were found in the gross scores among the medication,1,2,3 and 4 moxa-cone groups(P>0.05).The histopathological scores were significantly lower in the1 and 4 moxa-cone Moxi groups than in the medication group(P<0.05,P<0.01);and significantly lower in the 1,3,4 moxacone Moxi groups than in the 2 moxa-cone Moxi group(P<0.05,P<0.01).PAS staining showed a significant increase of the AOD values of colonic mucins in the 1,2,3 and 4 moxa-cone and medication groups relevant to the model group(P<0.01);and the AOD values of colonic mucins in the 1,3,4 moxa-cone Moxi groups were significantly increased than that in the 2 moxa-cone Moxi group(P<0.05,P<0.01).HID-AB staining showed that the AOD values of sulfated mucus content were significantly higher in the 2 and 4 moxa-cone Moxi groups than in the 3 moxa-cone Moxi group(P<0.01).The two-level two-factor factorial analysis showed an interaction existed between the moxa-cone number and Moxi frequency in reducing the gross score and histopathological score and in facilitating colonic mucin and sulfated mucus secretion.The histopathological score of the 4 moxa-cone Moxi group was significantly lower than that of the 2 moxa-cone Moxi group(P<0.05),and the sulfated mucus content was significantly higher in the 4 moxa-cone group than in the 3 moxa-cone group(P<0.01).The effect of Moxi given on alternate days was superior to that of daily Moxi in improving colonic histological damage.Conclusion Herbal cake-partitioned moxibustion at ST 25 can promote repair of the damaged colonic tissue and secretion of mucin in UC rats.The number of moxa cones and intervention frequency affect the efficacy of Moxi in improving histopathological changes.The Moxi intervention on alternate days and with 2 moxa-cones every time is recommended.
引文
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[6]刘兰英,雷玉婷,王和生.浅谈对艾灸灸量的认识[J].中国针灸,2015,35(11):1140-1142.
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[8]口锁堂,吴焕淦,刘慧荣,等.灸量的认识及意义[J].中华中医药学刊,2008,26(5):935-937.
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[13]史灵心,哈丽娟,曹方,等.现代针灸文献治疗腹痛的同功穴规律分析[J].世界中医药,2016,11(2):198-201.
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[15]包春辉,吴璐一,吴焕淦,等.针灸治疗活动期克罗恩病:随机对照研究[J].中国针灸,2016,36(7):683-688.
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[17]胥丽辉,王健,王锐,等.天枢穴治疗慢性便秘随机文献的应用规律分析[J].针灸临床杂志,2015,31(4):70-72.
[18]吴淑华.择时艾灸足三里、天枢穴促进食管癌术后胃肠功能恢复的探讨[J].光明中医,2015,30(4):805-807.
[19]李为贵,王彬彬,秦力.艾灸治疗功能性消化不良的临床观察[J].湖北中医杂志,2015,37(2):60.
[20]金焱.附子理中汤配合艾灸治疗原发性痛经65例[J].陕西中医,2010,31(3):280-281.
[21]陈德宇.健脾止泻汤联合隔药灸治疗溃疡性结肠炎随机平行对照研究[J].实用中医内科杂志,2013,27(20):15-17.
[22]刘瑛,谭振纹.隔药灸治疗慢性溃疡性结肠炎65例疗效观察[J].新中医,2007,39(12):21-22.
[23]吴焕淦,施征,朱毅,等.隔药灸治疗溃疡性结肠炎的临床研究[J].上海针灸杂志,2007,26(4):3-4.
[24]温凌洁.隔药灸治疗慢性溃疡性结肠炎临床观察[J].江西中医学院学报,2003,15(2):35-36.
[25]WANG Y,PARKER C E,FEAGAN B G,et al.Oral5-aminosalicylic acid for maintenance of remission in ulcerative colitis[J].Cochrane Database Syst Rev,2016(5):CD 000544.
[26]WU H G,ZHOU L B,SHI D R,et al.Morphological study on colonic pathology in ulcerative colitis treated by moxibustion[J].World J Gastroenterol,2000,6(6):861-865.
[27]JI J,LU Y,LIU H,et al.Acupuncture and moxibustion for inflammatory bowel diseases:a systematic review and metaanalysis of randomized controlled trials[J].Evid Based Complement Alternat Med,2013,2013:158352.
[28]张保球,杨洁仪.不同灸量治疗原发性骨质疏松症临床研究[J].上海针灸杂志,2015,34(6):568-569.
[29]苟朝琴,高静,吴晨曦,等.艾条灸灸量时间参数对原发性痛经疗效影响的Meta分析[J].中医临床研究,2016,8(34):14-20.
[30]付勇,章海凤,熊俊,等.热敏灸治疗肠易激综合征不同灸量的临床疗效观察[J].中国针灸,2014,34(1):45-48.
[31]范洪力,柯斌霞,卢淑洪,等.溃疡性结肠炎热敏灸灸温灸感观察研究[J].光明中医,2016,31(11):1604-1607.
[32]曾晓霞.不同灸量对慢性溃疡性结肠炎大鼠血清IL-8、IL-10含量影响的研究[D].沈阳:辽宁中医药大学,2014.
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[2]殷积美,宁瑞花,刘巍.溃疡性结肠炎患者生活质量的调查及分析[J].解放军护理杂志,2008,25(9A):40-41.
[3]JOOS S,WILDAU N,KOHNEN R,et al.Acupuncture and moxibustion in the treatment of ulcerative colitis:a randomized controlled study[J].Scand J Gastroenterol,2006,41(9):1056-1063.
[4]WU H,CHEN H,HUA X,et al.Clinical therapeutic effect of drug-separated moxibustion on chronic diarrhea and its immunologic mechanisms[J].J Tradit Chin Med,1997,17(4):253-258.
[5]LI H,HE T,XU Q,et al.Acupuncture and regulation of gastrointestinal function[J].World J Gastroenterol,2015,21(27):8304-8313.
[6]刘兰英,雷玉婷,王和生.浅谈对艾灸灸量的认识[J].中国针灸,2015,35(11):1140-1142.
[7]王桂英,王耀帅,王玲玲.艾灸疗法中灸感、灸温、灸量与灸效关系[J].中医杂志,2015,56(17):1519-1521.
[8]口锁堂,吴焕淦,刘慧荣,等.灸量的认识及意义[J].中华中医药学刊,2008,26(5):935-937.
[9]徐叔云,卞如濂,陈修.药理实验方法学[M].3版,北京:人民卫生出版社,2002:1335.
[10]WANG X,LIU Y,DING H,et al.Herb-partitioned moxibustion regulates the TLR 2/NF-κB signaling pathway in a rat model of ulcerative colitis[J].Evid Based Complement Alternat Med,2015,2015:949065.
[11]BUTZEN J D,PARMAR R,BELL C J,et al.Butyrate enema therapy stimulates mucosal repair in experimental colitis in the rat[J].Gut,1996,38(4):568-573.
[12]DIELEMAN L A,PALMEN M J,AKOL H,et al.Chronic experimental colitis induced by dextran sulphate sodium(DSS)is characterized by Th 1and Th 2cytokines[J].Clin Exp Immunol,1998,114(3):385-391.
[13]史灵心,哈丽娟,曹方,等.现代针灸文献治疗腹痛的同功穴规律分析[J].世界中医药,2016,11(2):198-201.
[14]黄琴峰.消化系统针灸疾病谱现代文献计量分析与评价[J].中国针灸,2007,27(10):769-777.
[15]包春辉,吴璐一,吴焕淦,等.针灸治疗活动期克罗恩病:随机对照研究[J].中国针灸,2016,36(7):683-688.
[16]王栋斌,葛淑琦,邱荃.艾灸天枢穴治疗脾胃虚弱型泄泻的临床观察[J].中国民间疗法,2016,24(6):15.
[17]胥丽辉,王健,王锐,等.天枢穴治疗慢性便秘随机文献的应用规律分析[J].针灸临床杂志,2015,31(4):70-72.
[18]吴淑华.择时艾灸足三里、天枢穴促进食管癌术后胃肠功能恢复的探讨[J].光明中医,2015,30(4):805-807.
[19]李为贵,王彬彬,秦力.艾灸治疗功能性消化不良的临床观察[J].湖北中医杂志,2015,37(2):60.
[20]金焱.附子理中汤配合艾灸治疗原发性痛经65例[J].陕西中医,2010,31(3):280-281.
[21]陈德宇.健脾止泻汤联合隔药灸治疗溃疡性结肠炎随机平行对照研究[J].实用中医内科杂志,2013,27(20):15-17.
[22]刘瑛,谭振纹.隔药灸治疗慢性溃疡性结肠炎65例疗效观察[J].新中医,2007,39(12):21-22.
[23]吴焕淦,施征,朱毅,等.隔药灸治疗溃疡性结肠炎的临床研究[J].上海针灸杂志,2007,26(4):3-4.
[24]温凌洁.隔药灸治疗慢性溃疡性结肠炎临床观察[J].江西中医学院学报,2003,15(2):35-36.
[25]WANG Y,PARKER C E,FEAGAN B G,et al.Oral5-aminosalicylic acid for maintenance of remission in ulcerative colitis[J].Cochrane Database Syst Rev,2016(5):CD 000544.
[26]WU H G,ZHOU L B,SHI D R,et al.Morphological study on colonic pathology in ulcerative colitis treated by moxibustion[J].World J Gastroenterol,2000,6(6):861-865.
[27]JI J,LU Y,LIU H,et al.Acupuncture and moxibustion for inflammatory bowel diseases:a systematic review and metaanalysis of randomized controlled trials[J].Evid Based Complement Alternat Med,2013,2013:158352.
[28]张保球,杨洁仪.不同灸量治疗原发性骨质疏松症临床研究[J].上海针灸杂志,2015,34(6):568-569.
[29]苟朝琴,高静,吴晨曦,等.艾条灸灸量时间参数对原发性痛经疗效影响的Meta分析[J].中医临床研究,2016,8(34):14-20.
[30]付勇,章海凤,熊俊,等.热敏灸治疗肠易激综合征不同灸量的临床疗效观察[J].中国针灸,2014,34(1):45-48.
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