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.
引文
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