Zhang Dan, Ren Yan-Bo, Wu Huan-Gan, Yang Yan-Ting, Wu Li-Jie, Zhang Ji, Shi Zheng, Ma Xiao-Peng
Shanghai Research Institute of Acupuncture-moxibustion and Meridian, Shanghai 200030, China.
Zhongshan Hospital Affiliated to Fudan University, Shanghai 201907.
Zhen Ci Yan Jiu. 2018 Feb 25;43(2):68-74. doi: 10.13702/j.1000-0607.170843.
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.
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 (=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 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 dia-mine-alcian blue (HID-AB) staining.
Compared with the normal group, rats in the model group presented loose stool, or with pus and blood, and slowly increased body weight (<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 (<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 (<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 (>0.05). The histopathological scores were significantly lower in the 1 and 4 moxa-cone Moxi groups than in the medication group (<0.05, <0.01); and significantly lower in the 1, 3, 4 moxa-cone Moxi groups than in the 2 moxa-cone Moxi group (<0.05, <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 (<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 (<0.05, <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 (<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 (<0.05), and the sulfated mucus content was significantly higher in the 4 moxa-cone group than in the 3 moxa-cone group (<0.01). The effect of Moxi given on alternate days was superior to that of daily Moxi in improving colonic histological damage.
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.
观察不同剂量隔药饼灸对溃疡性结肠炎(UC)大鼠受损结肠组织病理变化的影响,筛选隔药饼灸治疗UC的最佳剂量。
将63只雄性Sprague Dawley(SD)大鼠随机分为7组:正常对照组、模型组、药物组、1、2、3、4壮隔药饼灸组(每组9只)。采用将含结肠黏膜制备蛋白悬液与完全弗氏佐剂的乳剂(1 mL)皮下注射至大鼠足趾、腹股沟及背部的方法制备UC模型。于第38天,用3%甲醛及上述乳剂灌肠。将由附子、肉桂等组成的药饼置于双侧“天枢”(ST 25)穴进行隔药饼灸,每次1或2壮(每壮约5分钟),每日1次或隔日1次。观察大鼠一般情况(饮食、活动、反应能力、粪便及体重),苏木精-伊红(HE)染色后观察结肠组织病理变化(粘连、溃疡形成及炎症)并评分(组织病理学评分)。在立体显微镜下根据结肠组织粘连、溃疡形成及炎症的严重程度给予大体评分。过碘酸-希夫(PAS)染色后检测结肠黏蛋白的平均光密度(AOD)值,高铁二胺-阿尔辛蓝(HID-AB)染色后检测硫酸化黏液含量的AOD值。
与正常组相比,模型组大鼠出现稀便或脓血便,体重增长缓慢(<0.01),结肠明显充血,有溃疡斑或连续的浅表溃疡区,腺腔不规则,固有层和黏膜下层肿胀且炎症浸润严重,结肠组织损伤评分增加(<0.01)。PAS和HID-AB染色显示,模型组结肠黏蛋白和硫酸化黏液的AOD值与正常对照组相比显著降低(<0.01),提示肠腺黏液分泌减少。干预后,隔药饼灸组大鼠体重增加,粪便成形,受损结肠组织得到改善,黏膜愈合,炎症减轻,与模型大鼠相比损伤评分显著降低。药物组、1、2、3、4壮组的大体评分差异无统计学意义(>0.05)。1壮和4壮隔药饼灸组的组织病理学评分显著低于药物组(<0.05,<0.01);1、3、4壮隔药饼灸组的组织病理学评分显著低于2壮隔药饼灸组(<0.05,<0.01)。PAS染色显示,1、2、3、4壮组及药物组结肠黏蛋白的AOD值与模型组相比显著增加(<0.01);1、3、4壮隔药饼灸组结肠黏蛋白的AOD值显著高于2壮隔药饼灸组(<0.05,<0.01)。HID-AB染色显示,2壮和4壮隔药饼灸组硫酸化黏液含量的AOD值显著高于3壮隔药饼灸组(<0.01)。两因素析因分析显示,壮数与艾灸频率在降低大体评分和组织病理学评分以及促进结肠黏蛋白和硫酸化黏液分泌方面存在交互作用。4壮隔药饼灸组的组织病理学评分显著低于2壮隔药饼灸组(<0.05),4壮组硫酸化黏液含量显著高于3壮组(<0.01)。隔日艾灸在改善结肠组织学损伤方面的效果优于每日艾灸。
ST 25穴隔药饼灸可促进UC大鼠受损结肠组织的修复及黏蛋白分泌。灸壮数和干预频率影响隔药饼灸改善组织病理学变化的疗效。推荐隔日进行隔药饼灸干预,每次2壮。