| ObjectTo observe the general condition,daily behavioral changes,changes of gastrointestinal motility,Vimentin in rats with diarrhea-predominant IBS(IBS-D)model induced by electroacupuncture.To investigate the effects of cytoskeletal protein-vimentin expression on the contraction of gastrointestinal smooth muscle cells and the possible mechanisms of electroacupuncture in treating IBS-D rats with liver depression and spleen deficiency by regulating cytoskeletal protein expression.MethodThree 60-month-old SPF female Sprague-Dawley rats were randomly divided into 4 groups according to body weight: blank group,model group,electroacupuncture group,Pinaverium Bromide(PB)group,15 rats in each group.High-lactose diet fed with Chronic Unpredictable Mild Stress(CUMS)was used to prepare the rat model of IBS-D.After the successful modeling,electroacupuncture treatment was performed.The "Zusanli" and "Tianshu" points were selected and treated for 14 days,once a day.Observe the general conditions,changes in body weight,daily food intake,Bristol stool scores,sugar-water preference,gastric emptying,and intestinal propulsive rate of each group;Observe the morphological changes of colon tissue by HE staining;Immunohistochemistry The positive expression of Vimentin protein in the colonic mucosa was detected.The expression of Vimentin protein in the colon was detected by Western blot.Result1.Observe the behavior of rats in each group before and after treatment: After the modeling,the rats in each group were slow in movement,lying in bed,mental fatigue,dull hair,pale eyes and mucous membranes,pale ear,small reaction,and sound.Weak,loose stools are not forming,the color is yellow-green.After being treated with pinaverium bromide and electro-acupuncture for 14 days,the mental state of the rats was significantly better than that of the model.The activity was enhanced,the reaction of catching and repelling was strong,the hair was not disordered,the auricle was pale,and the faeces dried.Compared with the blank group,the scores of rats in each model group were significantly lower(P<0.05).Compared with the model group,the difference between the electroacupuncture group and the pinaverium bromide group was statistically significant(P<0.05).Significantly higher than the model group,and the Pinaverium bromide group score was significantly lower than the EA group(P<0.05);Compared with the blank group,the body weight growth rate of the rats in each model group decreased significantly with the increase of the time of modeling.After two weeks of treatment,the body mass growth rate has rebounded to varying degrees.In particular,after the intervention of electroacupuncture,the body mass growth rate was close to the level of the blank group;after the model was established,the daily food intake of the model group increased compared with the blank group.Significant difference(P<0.05),but after 14 days of treatment,the food intake in the EA group and the pinaverium bromide group was significantly increased compared with the model group(P<0.05),especially in the EA group compared with the pinaverium bromide group.Significant(P<0.05);Bristol stool scores before modeling were not significantly different in each group,and after 14 days of modeling,the Bristol score of the model group was significantly higher(P<0.05),after 14 days of treatment,the electroacupuncture group and The ratio of pinaverium bromide group was significantly lower than that of the model group(P<0.05).The group Bristol Bristol stool trait score was significantly lower than that of the pinaverium bromide group(P<0.05);the sugar and water preference of the blank group and the model group had a significant change before and after the modeling,and the model group was significantly lower than the blank group,the difference was statistically significant(P<0.05).2.Changes in gastrointestinal motility: compared with the blank group,the gastric emptying rate of the model group rats was significantly lower(P<0.05),while the small bowel propulsion rate was significantly higher(P<0.05);after treatment,electricity The gastric emptying rate in the acupuncture group was significantly higher than that in the model group and pinaverium bromide group(P<0.05),and was slightly lower than that in the blank group,suggesting that electroacupuncture treatment significantly improved the gastrointestinal motility of IBS.The pinaverium bromide group had a significant therapeutic effect on the intestinal propulsion rate of IBS rats(P<0.05),but it was still lower than that of the electroacupuncture group,and there was no significant improvement in gastric emptying.3.Effect on the histomorphology of rats in each group: In the blank group,the structure of the colon was normal,the mucous membrane was intact,the villi was arranged neatly,and the muscular layer was even and thick.In the model group,the villi were arranged irregularly or absent,and the muscularis became thinner.Compared with the model group,the electroacupuncture group and the pinaverium bromide group had uniform arrangement of villi,a single layer of columnar epithelium on the surface,a muscular layer in the middle lamina propria,and a slightly thickened submucosal muscle layer.The fibrous connective tissue in each group had no hyperplasia,no interstitial edema,and no infiltration of neutrophils,which was consistent with the characteristics of clinical IBS patients with no morphological abnormalities of the colon.The results of immunohistochemistry and immunoblotting of colon tissue in each group:Vimentin was mainly expressed in cytoplasm and close to cell membrane.The positive expression of Vimentin was light yellow or brown-yellow particles.Vimentin protein expression is low in normal tissues,and low expression of Vimentin in the submucosa muscle layer is of great significance to gastrointestinal smooth muscle contraction.The expression of Vimentin protein in the model group was significantly higher than that in the blank group,and the expression of Vimentin protein in the electroacupuncture group was significantly decreased(P<0.05),while the Pinaverium bromide group was not statistically significant(P>0.05);compared with the blank group,the expression of Vimentin protein in the model group did not change significantly.After treatment with pinaverium bromide,Vimentin protein expression was significantly reduced.Especially after treatment with electroacupuncture,Vimentin protein showed a low expression status.ConclusionUsing high-lactose diet feed,combined with chronic unpredictable mild stimulation method to successfully prepare IBS-D rat model of liver depression and spleen deficiency,electroacupuncture to improve the gastrointestinal motility of rats with irritable bowel syndrome,electroacupuncture "Zusanli" Acupoints can down-regulate the content of Vimentin protein and thus have a good regulatory effect on smooth muscle cell contraction and improve gastrointestinal dysfunction in IBS rats. |
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