溃结宁膏穴位贴敷对脾肾阳虚型溃疡性结肠炎大鼠的治疗作用及机制研究OA

Objective To observe the therapeutic effect of Kuijiening Ointment acupoint application on rat models with ulcerative colitis(UC)of spleen-kidney yang deficiency syndrome,and to explore its regulatory effects on intestinal inflammation,intestinal barrier function and intestinal flora.Methods Total 50 rats were randomly selected from 60 male SPF-grade SD rats.Rat models of spleen-kidney yang deficiency syn-drome were established by intragastric administration of adenine and Fanxieye(Sennae Folium)combined with cold-damp environment stimulation,and UC was induced by 2,4,6-trinitrobenzene sulfonic acid(TNBS)enema to construct a composite model with UC of spleen-kidney yang deficiency syndrome.The successfully modeled rats were randomly divided into 5 groups:the model group,acupoint application group,non-acupoint application group,acupoint placebo application group and sulfasalazine(SASP)group,with the remaining non-modeled rats serving as the blank group.All groups were intervened continuously for 14 days.The blank group and model group were given normal saline by gavage.The acupoint application group received Kuijiening Oint-ment application at specific acupoints and normal saline gavage.The non-acupoint application group was treated with Kuijiening Ointment applied to the non-acupoint area of the buttocks and normal saline gavage.The acu-point placebo application group was given blank cataplasm application at specific acupoints and normal saline gavage.The SASP group was administered SASP by gavage.After continuous intervention,the disease activ-ity index(DAI)and colonic tissue damage index(TDI)of rats in each group were evaluated,and the pathologi-cal changes of colonic tissues were observed.The levels of tumor necrosis factor-α(TNF-α)and interleukin-4(IL-4)in colonic tissues,as well as the expression of tight junction proteins(ZO-1,Occludin)were detected.The diversity of intestinal flora and the changes in the abundance of dominant phyla were analyzed by 16S rRNA gene sequencing technology,followed by comparative analysis.Results Compared with the blank group,the colonic length of rats in the model group was significantly shortened,the levels of DAI,TDI and TNF-α in colonic tissues were significantly increased(all P<0.01),the body weight and IL-4 level were significantly decreased(P<0.01),the expression of ZO-1 and Occludin proteins in colonic tissues was significantly reduced(P<0.05,P<0.01),the diversity of intestinal flora was markedly decreased,the relative abundance of Fir-micutes was reduced(P<0.05),and the relative abundance of Bacteroidetes was increased(P<0.01).Com-pared with the model group,the colonic length of rats in the acupoint application group and SASP group was significantly longer,the levels of DAI,TDI and TNF-α in colonic tissues were significantly lower(all P<0.01),the body weight,IL-4 level and Occludin protein expression in colonic tissues were significantly increased(P<0.01),the expression of ZO-1 protein was also elevated(P<0.05 or P<0.01),the diversity of intestinal flora was improved,the relative abundance of Firmicutes was increased(P<0.05,P<0.01),and the relative abundance of Bacteroidetes was decreased(P<0.01).Conclusion Acupoint application of Kui-jiening Ointment may exert the therapeutic effect on UC of spleen-kidney yang deficiency syndrome through multiple pathways,including inhibiting inflammatory response,repairing intestinal mucosal barrier and regulat-ing the balance of intestinal flora.