Moxibustion down-regulates colonic epithelial cell apoptosis and repairs tight junctions in rats with Crohn's disease
Moxibustion down-regulates colonic epithelial cell apoptosis and repairs tight junctions in rats with Crohn's disease
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DOI：
10.3748/wjg.v17.i45.4960
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发表时间：
2011-12-07
影响因子：
4.3
通讯作者：
Wang, Jin-Hai
中科院分区：
文献类型：
--
作者：
Bao, Chun-Hui;Wu, Lu-Yi;Wang, Jin-Hai
目的：目的：观察艾灸对克罗恩病（CD）大鼠结肠上皮细胞凋亡及紧密连接修复的影响。将60只雄性SD大鼠随机分为正常对照组（NC）、模型对照组（MC）、隔药灸组（HPM）、温灸组（MWM）和柳氮磺胺吡啶组（SASP）。每组12只大鼠。用三硝基苯磺酸诱导CD模型大鼠肠道炎症反应。HPM组和MWM组大鼠分别于天枢、气海穴治疗1次/d，连续14 d; SASP组给予SASP 2次/d，连续14 d。MC组和NC组不给予额外治疗。透射电镜下观察结肠上皮的显微结构，短路电流法测定跨上皮电阻，末端脱氧核苷酸转移酶介导的dUTP-生物素缺口末端标记法测定结肠上皮细胞凋亡，免疫印迹法和免疫荧光染色法检测结肠上皮连接部claudin-1和闭合小带ZO-1的表达。与MC组相比，HPM、MWM和SASP组大鼠结肠上皮屏障的微观结构明显改善，电流明显减少，数值分别为168.20 +/- 6.14 vs 99.70 +/- 3.13、99.10 +/- 4.28和120.30 +/- 3.65 mA（P = 0.001）。然而，HPM和MWM组的电流流速高于SASP组（99.70 +/- 3.13，99.10 +/- 4.28 vs 120.30 +/- 3.65 mA，P = 0.001）。HPM、MWM和SASP组结肠上皮细胞凋亡数量明显减少（分别为61.5 ± 16.91 vs 15.5 ± 8.89、14.8 ± 6.27和24.7 ± 9.68（P = 0.001）;封闭蛋白、封闭蛋白-1和ZO-1在MWM组和HPM组的表达均明显增强（对于闭合蛋白为0.48 +/- 0.10，0.64 +/- 0.09对0.18 +/- 0.05，对于闭合蛋白-1为0.12 +/- 0.02，0.17 +/- 0.03对0.05 +/- 0.01，以及对于闭合蛋白-1为0.08 +/- 0.01，0.11 +/- 0.01对比ZO-1的0.02 +/- 0.01）。在SASP组中，occludin和ZO-1的表达也显著增加（occludin为0.27 ± 0.04 vs 0.18 ± 0.05，ZO-1为0.05 ± 0.01 vs 0.02 ± 0.01），但claudin-1无显著差异。结论：HPM和MWM能下调CD大鼠结肠上皮细胞凋亡，修复紧密连接，增强结肠上皮屏障功能。(C)2011年百世登。All rights reserved.
AIM: To investigate the effects of moxibustion on down-regulation of the colonic epithelial cell apoptosis and repair of the tight junctions in rats with Crohn's disease (CD).METHODS: Sixty male Sprague-Dawley rats were randomly divided into a normal control (NC) group, a model control (MC) group, an herbs-partitioned moxibustion (HPM) group, a mild-warm moxibustion (MWM) group and a salicylazosulphapyridine (SASP) group, with 12 rats in each group. The CD model rats were treated with trinitrobenzene sulphonic acid to induce intestinal inflammation. The rats in the HPM and MWM groups were treated at the Tianshu (ST25) and Qihai (CV6) acupoints once daily for 14 d, and the SASP group was fed SASP twice daily for 14 d. No additional treatment was given to the MC and NC groups. The microstructure of the colonic epithelium was observed under a transmission electron microscope, the transepithelial resistance was measured using a short-circuit current, colonic epithelial cell apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling assay, and the expression of occludin, claudin-1 and zonula occludens-1 (ZO-1) in the colonic epithelial junction was determined by Western blotting and immunofluorescence staining.RESULTS: Compared with the MC group, the microstructure of the colonic epithelial barrier was significantly improved in rats treated with HPM, MWM or SASP, meanwhile, the current flow was reduced significantly, with values of 168.20 +/- 6.14 vs 99.70 +/- 3.13, 99.10 +/- 4.28 and 120.30 +/- 3.65 mA, respectively (P = 0.001). However, the HPM and MWM groups had higher current flow rates than the SASP group (99.70 +/- 3.13, 99.10 +/- 4.28 vs 120.30 +/- 3.65 mA, P = 0.001). The number of the apoptotic colonic epithelial cells in HPM, MWM and SASP groups was largely reduced (61.5 +/- 16.91 vs 15.5 +/- 8.89, 14.8 +/- 6.27 and 24.7 +/- 9.68, respectively (P = 0.001); and the expression of occludin, claudin-1 and ZO-1 in the MWM and HPM groups was significantly enhanced (0.48 +/- 0.10, 0.64 +/- 0.09 vs 0.18 +/- 0.05 for occludin, 0.12 +/- 0.02, 0.17 +/- 0.03 vs 0.05 +/- 0.01 for claudin-1, and 0.08 +/- 0.01, 0.11 +/- 0.01 vs 0.02 +/- 0.01 for ZO-1). And in SASP group, the expression of occludin and ZO-1 was also significantly increased (0.27 +/- 0.04 vs 0.18 +/- 0.05 for occludin and 0.05 +/- 0.01 vs 0.02 +/- 0.01 for ZO-1), but there was no significant difference for claudin-1. The HPM and MWM groups had higher expression of occludin, claudin-1 and ZO-1 than the SASP group.CONCLUSION: HPM and MWM treatment can downregulate apoptosis of colonic epithelial cells, repair tight junctions and enhance colonic epithelial barrier function in rats with CD. (C) 2011 Baishideng. All rights reserved.