title: Moxibustion down-regulates colonic epithelial cell apoptosis and repairs tight junctions in rats with Crohn's disease.
abstract: OBJECTIVE
  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-l (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.
  
  
  CONCLUSIONS
  HPM and MWM treatment can down-regulate apoptosis of colonic epithelial cells, repair tight junctions and enhance colonic epithelial barrier function in rats with CD.
authors.affiliation: Shanghai Institute of Acupuncture-Moxibustion and Meridian, Shanghai University of Traditional Chinese Medicine, Shanghai 200030, China.
authors.forename: Chun-Hui
authors.initials: CH
authors.lastname: Bao
authors.forename: Lu-Yi
authors.initials: LY
authors.lastname: Wu
authors.forename: Yin
authors.initials: Y
authors.lastname: Shi
authors.forename: Huan-Gan
authors.initials: HG
authors.lastname: Wu
authors.forename: Hui-Rong
authors.initials: HR
authors.lastname: Liu
authors.forename: Rong
authors.initials: R
authors.lastname: Zhang
authors.forename: Li-Qing
authors.initials: LQ
authors.lastname: Yu
authors.forename: Jin-Hai
authors.initials: JH
authors.lastname: Wang
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D062445:Claudin-1; D003106:Colon / Q000166:cytology / Q000473:pathology*; D003424:Crohn Disease / Q000473:pathology* / Q000628:therapy*; D015536:Down-Regulation; D004847:Epithelial Cells / Q000502:physiology* / Q000648:ultrastructure; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology / Q000473:pathology; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D009071:Moxibustion; D062793:Occludin; D010750:Phosphoproteins / Q000378:metabolism; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley; D019108:Tight Junctions / Q000378:metabolism*; D062826:Zonula Occludens-1 Protein
chemical_list: C568108:CLDN1 protein, human; D062445:Claudin-1; C568114:Cldn1 protein, rat; D008565:Membrane Proteins; C568985:OCLN protein, human; D062793:Occludin; C568987:Ocln protein, rat; D010750:Phosphoproteins; C569042:TJP1 protein, human; C569044:Tjp1 protein, rat; D062826:Zonula Occludens-1 Protein