| ObjectIrritable bowel syndrome(IBS)is a common functional gastrointestinal disease accompanied by typical clinical symptoms such as abdominal pain,discomfort,changes in bowel habits,and abnormal stool characteristics.The etiology and pathogenesis of this disease are not yet fully understood,and the impaired intestinal mucosal barrier function may be one of the main factors.As an important part of the intestinal mucosal barrier,the tight junction structure is distributed at the top and edge of the intestinal mucosal cell membrane.Most patients with IBS have abnormalities and permeability changes in the intestinal mucosal tight junction structure in clinical practice,suggesting that the above structure may be related to The incidence of IBS is related.Therefore,this experiment uses a combination of acute stress and chronic stress to prepare an IBS-D rat model,and uses electroacupuncture to intervene while building the model.Starting from the intestinal mucosal barrier mechanism,the tight junction structure is the starting point.To observe the effects of electroacupuncture at different acupoints on the behavior,intestinal epithelial mucosal tissue morphology and related protein expression in IBS-D rats,clarify the protective effect of electroacupuncture on the tight junction structure,and hope to provide clinical treatment for irritable bowel syndrome Experimental basis.MethodThree-month-old SPF male SD rats with a total of 40 and weighing180-200 g were used.After one week of adaptive feeding,they were divided into four groups by random number table method,each with10 rats,namely: blank group,Model group,combined point matching group,conventional point matching group.The IBS-D rat model was prepared by combining acute stress and chronic stress,and the modeling time was 28 days.Electro-acupuncture intervention was performed on rats while modeling.The blank group was fed ordinary feed without other treatments;the model group received daily grasping training and rat clothes putting on and taking off training while modeling;the group of co-recruiting acupuncture points At the same time,the model was created with electro-acupuncture at Zusanli and Tianshu points on both sides,continuous wave 2HZ,current intensity 1m A,treatment once a day,15 minutes each time;the conventional acupuncture group used acupuncture on both sides at the same time as the model was created.Treatment of Zusanli and Shangjuxu points,continuous wave 2HZ,current intensity 1m A,treatment once a day,15 minutes each time.After modeling and electroacupuncture intervention,observe the general condition and physical signs of rats,and observe the morphological changes of rat colon tissue by HE staining method;immunohistochemical method(Immunohistochemical)detects the positive of colon tissue tight junction proteins Occuldin and Claudin-1 Expression;Western blot was used to detect the expression levels of Occuldin and Claudin-1proteins in the colon.Result1.Observation of general condition and body weight of rats in each groupIn terms of the general conditions of rats,the blank group had no significant changes in mental state,activity,hair gloss and fecal traits before and after modeling;the model group showed fatigue,sluggishness,withered hair and loose stools in the middle and late stages of modeling.The dirty and smelly condition indicates that the model is more successful;the general condition of the combined point matching group and the conventional point matching group is significantly improved in the middle and late stage of modeling + intervention,and the spirit becomes better,and the fecal traits gradually become dry from loose and dry,prompting electroacupuncture Intervention can improve the general condition of rats.In terms of the body weight of rats,compared with the blank group,the growth rate of body weight of rats in the model group,the combined acupoint group and the conventional acupoint group has slowed down after the start of modeling,and the difference was statistically significant(P<0.05));Compared with the model group,in the middle and late stages of model building + intervention,the weight of the combined acupoint matching group and the conventional acupoint matching group increased significantly,and the difference was statistically significant(P<0.05);compared with the conventional acupoint matching group In comparison,the weight gain of the co-recruiting acupoints group was better at the late stage of model building + intervention,and the difference was statistically significant(P<0.05).2.Comparative observation of Bristol score,sugar water preference result,and AWR score of each group of ratsIn terms of rat Bristol scores,compared with the blank group,the Bristol scores of rats in the other groups increased after the start of modeling,and the difference was statistically significant(P<0.05);compared with the model group,electroacupuncture intervention The Bristol scores of the two groups of rats were significantly reduced,and the difference was statistically significant(P<0.05);while the combined acupoint matching group and the conventional acupoint matching group were compared,the difference was not statistically significant(P>0.05).In the comparison of the results of the sugar water preference experiment,there was no significant difference in the SP value of sugar water of all groups of rats before modeling.Compared with the blank group,the SP value of sugar water in the model group,the combined acupoint group and the conventional combined acupoint group was significantly lower,and the difference was statistically significant(P<0.05);compared with the model group,the combined acupoint group The SP value of rats increased after the intervention of electroacupuncture with the conventional point matching group,and the difference was statistically significant(P < 0.05);compared with the conventional point matching group,the SP value of the combined point matching group was relatively higher,the difference was There is statistical significance(P<0.05).In terms of AWR score,when the pressure is 20 mm Hg,compared with the blank group,the AWR scores of the other three groups have a more significant increase,the difference is statistically significant(P<0.05);when the pressure is 40 mm Hg,it is compared with the model group In comparison,the AWR scores of the combined-point combination group and the conventional combination-point group undergoing electroacupuncture intervention were reduced,and the difference was statistically significant(P<0.05);when the pressure was 60 mm Hg,the combined-point combination group and the conventional combination group had lower AWR scores.Compared with the model group,the AWR score of the acupoint group was reduced,and the difference was statistically significant(P<0.05);when the pressure was 80 mm Hg,the AWR scores of the rats in each group were compared,and the difference was not statistically significant(P> 0.05).3.Observation of the morphology of intestinal mucosa of rats in each groupIn the blank group,the colonic mucosa structure was complete,the villi were arranged in a neat manner,there was no obvious breakage,the capillaries were clear,and no inflammatory cell infiltration was found.In the model group,the colon tissue was damaged,the epithelial structure was obviously destroyed,the villi were arranged in disorder,the muscle layer became thin,and there was no edema in the lower layer of the mucosa.Compared with the model group,the combined point matching group and the conventional matching point group have a more complete colonic mucosa structure,a small amount of villi can still fall off,and slight breakage can be seen.The muscular layer can be seen in the middle lamina propria and the submucosal muscularis Thickness increases.However,no inflammatory cell infiltration and edema occurred in each group.4.Observation of Occludin and Claudin-1 protein expression in colon tissue of rats in each groupIn terms of immunohistochemical results,compared with the blank group,the expression of Occludin and Claudin-1 in the colon tissue of the remaining groups was significantly less,and the difference was statistically significant(P<0.05);compared with the model group,the expression of Occludin and Claudin-1 The positive expression of protein in the combined acupoint matching group and the conventional acupoint matching group of acupuncture treatment was significantly increased,and the difference was statistically significant(P<0.05);compared with the conventional acupoint matching group,the positive expression in the combined acupoint matching group was obvious More,the difference was statistically significant(P<0.05).In terms of Western blot results,compared with the blank group,the expression of Occludin and Claudin-1 protein in the model group and the ratio of GAPDH to GAPDH were significantly reduced(P<0.05);compared with the model group,the combined acupoint group after the intervention of electroacupuncture The protein expressions of Occludin and Claudin-1 in the group with conventional points were significantly increased,and the difference was statistically significant(P<0.05).Compared with the conventional acupoint matching group,there were significantly more positive expressions in the combined acupoint matching group,and the difference was statistically significant(P<0.05).ConclusionElectro-acupuncture at Zusanli and Tianshu points can improve the general condition and depression of IBS-D rats,regulate visceral hypersensitivity,up-regulate the expression of Occludin and Claudin-1 protein in rat colon tissue,repair tight junction structure,and improve intestinal mucosal barrier Features. |
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