Effects Of Direct Moxibustion On Colonic Epithelium And Expression Of TLR9-NF-κB In Ulcerative Colitis Rats

Purpose：The objective of this study is to investigate and to comparethe efficacy of moxibustion in the treatment of ulcerative colitis (UC)rats from morphological, immunological and molecular biologicalperspectives, further providing experimental support for clinicalapplications.Material and method:50Sprague-Dawley rats were randomly assignedto blank control group (normal rats, n=6; BC group) and model replicationgroup (UC rats, n=44; MR group). Ulcerative Colitis (UC) model wasestablished by enema of2,4,6-trinitrobenzenesulfonic acid(TNBS)/dextran sulfate sodium (DSS). Rates in MR group were furtherrandomly assigned to local point9-min moxibustion group (9moxa-cone,n=6;9LM group), local point6-min moxibustion group (6moxa-cone, n=6;6LM group), local point3-min moxibustion group (3moxa-cone, n=6;3LMgroup), meridian point9-min moxibustion group (9moxa-cone, n=6;9MMgroup), meridian point6-min moxibustion group (6moxa-cone, n=6;6MMgroup), meridian point3-min moxibustion group (3moxa-cone, n=6;3MMgroup), and waiting list control group (no moxibustion treatment, n=6;WLC group). Rats in moxibustion treatment group were treated in14sessions over28days. The disease activity index (DAI), local tissuemorphology, the levels of interleukin8(IL-8) as well as interleukin(IL-10) in the serum, and the expressions of toll-like receptor9(TLR9)as well as nuclear factor kappa-light-chain-enhancer of activated B cellsP65(NF-κB P65) in colonic tissue were determined by DAI scores, HEstaining, electronic microscopy, enzyme-linked immunosorbent assay andwestern-blot, respectively. Results：1. After moxibustion treatment, the DAI rating in9LM/9MM was lowest, andthat in WLC was highest. The differences of DAI rating between UC ratswith moxibustion treatment (9LM,6LM,3LM,9MM,6MM,3MM) and UC rats withoutmoxibustion treatment (WLC) were significant (P<0.05). From opticalmicroscopy and electronic microscopy results, we found that the neatnessof glandular arrangement in colonic mucosal epithelia of rats from WLC,3LM/3ML,6LM/6MM, to9LM/9MM gradually increased.Among those, the resultsfrom9LM and6LM were similar: relatively intact and ordered microvilli,tight cell junction, and even cellular matrix. The neatness/intactnessof microvilli is slightly improved in9MM, compared to6MM. In contrast,colonic mucosal epithelia of rats in3LM/3MM showed contained damagedmicrovilli with uneven length, loose cell junction, and unclear dissolvedmucous granule in the cytoplasm2. After moxibustion treatment, IL-8level successively decreased whilethe IL-10level increased for rats from WLC,3LM/3MM,6LM/6MM, to9LM/9MMgroups. For the expressions of both TLR9and NF-κB P65, both of themdecreased in order: the expression amounts in9LM/9MM were smallest andthose in WLC were greatest. In addition, the differences among differentgroup were mutually significant (P<0.05).Conclusion：1. Moxibustion therapy is effective in improving the disease activity ofUC rats, in terms of increasing body mass, improving fecal viscosity, andreducing rectal bleeding.The results from optical and electronicmicroscopy indicate: moxibustion can repair the damaged colonic mucosaand glandular arrangement; moxibustion can repair damaged microvilli andreduced unclear dissolved mucous granule; the more amount of moxibustionis applied, the better treatment effect is obtained.2. Moxibustion treatment is capable of suppressing the secretion of serum IL-8while activating that of IL-10, inhibiting the activation of NF-κBP65, and decreasing the expressions of TLR-9in UC rats. We have proposedthat moxibustion therapy effect on UC rats is probablydirectly/indirectly related to the inhibition of inflammatory cellsreleasing by suppressing the secretion of proinflammatory cytokine (IL-8)while activating the secretion of anti-inflammatory molecule (IL-10), andthe blocking of inflammatory signaling transduction pathway by inhibitingthe activation of transcription factor (NF-κB P65) as well as depressingthe pattern-recognition receptors (TLR9).3. After investigating and comparing the efficacy of moxibustion in thetreatment of UC rats from morphological, immunological and molecularbiological perspectives, we found that the therapy efficacy from localpoint groups is better than that from meridian point groups, given thesame condition.For example, treatment parameters from9LM give the bestresults, followed in decreasing order by6LM,9MM,3LM,6MM, and3MM.