隔药灸不同灸量对溃疡性结肠炎大鼠结肠组织病理学影响的实验研究
张 丹11 任燕波2 吴焕淦1 杨延婷3 吴丽洁3 张 霁3 施 征1 马晓芃1,3
(1上海市针灸经络研究所,上海 200030;2复旦大学附属中山医院,上海201907;3上海中医药大学,上海 20120)
摘要:研究目的:观察隔药灸不同灸量对溃疡性结肠炎 (Ulcerative colitis, 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大鼠UC的最佳灸量组合。研究方法:Sprague Dawley (SD) 大鼠随机分为7组,正常组、模型组、西药组、隔药灸1-4组,每组9只。采用抗原免疫加局部化学刺激复合法制备实验性UC模型。隔药灸1-4组均选取双侧“天枢”穴,基于施灸壮数和频次给予不同灸量组合隔附子饼灸治疗。观察大鼠一般情况,采用苏木素-伊红 (Hematoxylin-eosin, HE) 染色、过碘酸雪夫 (Periodic acid-schiff, PAS) 染色、高铁二胺阿辛蓝 (High iron diamine-alcian blue, HID-AB) 染色,研究结肠组织病理学变化。研究结果:与正常组比较,模型组大鼠稀便,部分脓血,体重增加缓慢 (P<0.01);巨检显示该组大鼠结肠充血明显,严重者局部存在数个白色溃疡点,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染色显示结肠肠腺粘液明显减少(P<0.01)。与模型组比较,隔药灸各组大鼠体重增加,大便成形,结肠组织损伤改善,表现为黏膜愈合和炎症减轻,损伤评分明显降低(P<0.05)。PAS染色、HID-AB染色显示隔药灸各组大鼠结肠腺体分泌粘蛋白增加(P<0.01)。两因素两水平析因分析结果显示,隔药灸壮数与频次对结肠组织评分、粘液分泌均有交互作用,共同影响灸效(P<0.05)。结论:隔药灸天枢穴具有促进UC大鼠结肠损伤修复及粘液分泌作用;从改善组织病理学角度,隔药灸施灸壮数与频次影响灸效,隔药灸隔日2壮灸为最佳灸量组合。
关键词:针灸;隔药灸;溃疡性结肠炎;苏木素-伊红染色;过碘酸雪夫染色;高铁二胺阿辛蓝染色;灸量
Histopathological Effects of Different-dose Herb-partitioned Moxibustion on Colon Tissue of Ulcerative Colitis Rats
ZHANG Dan1, REN Yanbo2, WU Huangan1, YANG Yanting3, WU Lijie3, ZHANG Ji3, SHI Zheng1, MA Xiaopeng1,3
(1Shanghai Research Institute of Acupuncture and Meridian, Shanghai 200030, China; 2 Zhongshan Hospital Affiliated to Fudan University, Shanghai 201907; 3 Shanghai University of Traditional Chinese Medicine, Shanghai 20120)
Abstract: Objective: To observe the effects of different-dose herb-partitioned moxibustion (HPM) on colon damage in rats with ulcerative colitis (UC), for selecting the optimal dosage of HPM from the histopathological view. Methods The Sprague Dawley (SD) rats were randomized into 7 groups, a normal group, a model group, a Western medication group, and HPM group 1-4, 9 rats per group. UC model was developed by antigen immune plus local chemical stimulation. In the four HPM groups, bilateral “Tianshu” (ST 25) were selected to receive corresponding dosage of HPM treatment varied at cone number and frequency. The general conditions of rats were observed. The pathological changes of colon tissues were examined by using hematoxylin-eosin (HE) staining, periodic acid-schiff (PAS) staining, and high iron diamine-alcian blue (HID-AB) staining. Results Compared with the normal group, rats in the model group presented loose stool, some with pus and blood, and slowly increased body weight (P<0.01); gross examination showed that rat colon in the model group had obvious congestion, a couple of white ulcer spots in severe cases; microscopic examination found continuous superficial ulcers in colon, with irregular glandular cavities, swelling and serious inflammatory infiltration in lamina propria and submucosa, and the score of colon tissue damage increased significantly (P<0.01); PAS and HID-AB staining demonstrated that the production of colonic mucus decreased markedly (P<0.01). Compared with the model group, rats in the HPM groups presented increased body weight, formed feces, and improved colon damages, manifested by healed mucosa and mitigated inflammation, and the damage score dropped significantly (P<0.05). PAS and HID-AB staining showed that the production of colonic mucins increased significantly in the HPM groups (P<0.01). The two-level two-factor factorial analysis indicated that both HPM cone number and frequency interacted with colon tissue score and the production of mucus, affecting the efficacy of moxibustion (P<0.05). Conclusion HPM at Tianshu (ST 25) promotes the repair of colonic damage and mucus production; from the histopathological view, the number of moxa cone and intervention frequency affect the efficacy of HPM, and HPM once every other day with 2 cones each time should be the optimal choice of dosage.
Key words: Acupuncture and moxibustion, Herb-partitioned moxibustion, Ulcerative colitis, HE staining, PAS staining, HID-AB staining, Moxibustion dosage