| Objective:To observe the effects of acupoint catgut embedding,acupoint moxibustion and their combined intervention on the morphology of colonic mucosa,colonic mucosal Notch1 receptor,target genes Hes1,and Math1 m RNA and protein expression in colonic mucosa of ulcerative colitis(UC)rats.To explore its role in the treatment of UC,and explore its possible mechanism from the perspective of Notch signaling pathway.Methods:Six sterile Sprague-Dawley(SD)rats were randomly selected from a group of 32 SD rats as normal group,and the remaining were prepared for UC modeling by intrarectal administration of 5% trinitro-benzene-sulfonic acid and 50% ethyl alcohol.UC model rats were randomly divided into four groups: model,acupoint catgut embedding combined with moxibustion,acupoint catgut embedding,and moxibustion groups(n=6 in each group).Bilateral “Shangjuxu”(ST37),“Tianshu”(ST25)and “Dachangshu”(BL25)were selected for treatment from the next day after grouping.Self-made moxa strips mild moxibustion on acupoints was applied for10 min,once a day for 14 days in moxibustion group.Simple embedding needles into acupoints were treated once a week for two weeks in acupoint catgut group.In the combined treatment group,moxibustion intervention was performed first,followed by embedding after 6 hours.The treatment method was the same as that of the moxibustion group and the acupoint catgut group.Disease activity index(DAI)was observed.Histological changes of colonic mucosa were observed by HE staining under light microscope;Protein and m RNA expression of Notch1,Hes1 and Math1 were detected by Western blot and q PCR,respectively.Results:1.General state and DAIThe rats in the normal group were in good mental condition,with shiny hair,normal diet and drinking water,granular stools,and no abnormal secretions around the anus.Model group rats were listless,lazy,yellow hair,significantly reduced diet and water consumption,thin and unformed stools,or accompanied by mucous pus and blood stool,perianal filth,and emaciation.Compared with the normal group,DAI of model group was significantly higher(P<0.01);The symptoms of rats in acupoint catgut group,moxibustion group and acupoint catgut embedding combined with moxibustion group improved to varying degrees,and their DAI decreased(P<0.05 or P<0.01).DAI of the acupoint catgut embedding combined with moxibustion group was significantly lower than the acupoint catgut embedding and moxibustion group(P<0.01).2.Histomorphological changes in colonic mucosaThe structure of colonic mucosal epithelium in the normal group was complete and clear,with abundant goblet cells and complete & orderly glandular ducts.The structure of the colonic mucosal epithelium in the model group was fuzzy,with a large number of damaged and exhumed epithelium,reduced goblet cells,damaged glands,a large number of inflammatory cell infiltration,congestion and edema in the mucosal layer.Intestinal mucosal epithelium of rats in acupoint catgut embedding group and moxibustion group was damaged shallowly,goblet cells increased,glands increased and arranged neatly,and inflammatory cells reduced.Intestinal mucosal epithelium of rats in the catgut embedding plus moxibustion group was not obvious damage,goblet cells increased,glands increased and arranged neatly,and inflammatory cells reduced significantly.3.Protein and m RNA expression levels of Notch1 receptor in colonic mucosaCompared with the normal group,expression of Notch1 receptor m RNA and protein in the colonic mucosa tissue of the rats in the model group were significantly increased(P<0.01).Compared with the model group,m RNA and protein expression levels of Notch1 receptor in all treatment groups were significantly decreased(P <0.01).Compared with the acupoint catgut embedding group and moxibustion group,m RNA and protein expression levels of Notch1 receptor decreased significantly in embedding plus moxibustion group(P<0.01).4.Expression levels of Hes1 m RNA and protein in colonic mucosaCompared with the normal group,m RNA and protein expressions of Hes1 in the colonic mucosa tissue of the rats in the model group were significantly higher(P<0.01).Compared with the model group,m RNA and protein expression levels of Hes1 in all treatment groups were decreased in different degrees(P<0.05 or P<0.01).Compared with the acupoint catgut embedding group and moxibustion group,the m RNA and protein expression levels of Hes1 receptor decreased significantly in embedding plus moxibustion group(P<0.01).5.Expression levels of Math1 m RNA and protein in colonic mucosaCompared with the normal group,m RNA and protein expressions of Math1 in the colonic mucosa tissue of the rats in the model group were significantly decreased(P<0.01).Compared with the model group,m RNA and protein expression levels of Math1 were significantly increased in the acupoint catgut embedding and acupoint catgut embedding combined with moxibustion group(P<0.01).m RNA expression level of Math1 in the moxibustion groups reflects no significant difference with model group(P>0.05),though with higher protein expression level(P<0.01).Compared with the acupoint catgut embedding combined with moxibustion group,there was no significant difference in the expression level of Math1 m RNA in acupoint catgut embedding group(P > 0.05),though protein expressions levels of Math1 were significantly decreased(P<0.01).m RNA and protein expressions of Math1 in the moxibustion groups were significantly increased comparing with moxibustion group(P<0.01).Conclusions:1.Acupoint catgut embedding,moxibustion and acupoint catgut embedding plus moxibustion can significantly relieve inflammation of intestinal mucosa and alleviate damage on intestinal mucosa in UC rats.The improvement effect of acupoint catgut embedding combined with moxibustion is better than acupoint catgut embedding or acupoint moxibustion,reflecting the synergistic effect of two therapies listed above.2.The therapeutical effect of acupoint catgut embedding,moxibustion and acupoint catgut embedding plus moxibustion of on UC may be realized by down-regulating the expression of Notch1 receptor with target gene Hes1 and up-regulating the expression of target gene Math1.Those therapies rebalance Hes1 and Math1,thus inhibiting the over-activation of Notch signaling pathway. |
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