| Objective: To observe the effect of electroacupuncture at Zusanli and Neiguan Points on the intestinal propulsive rate and 5-hydroxytryptamine(5-HT)and regeneration protein 3(regeneration protein 3)in duodenum of rats with middle cerebral artery occlusion(MCAO)at different time points(1D,3D,7D,14D)To explore the effect of Electroacupuncture on duodenal motility disorder in acute MCAO rats.Methods: 120 8-week-old healthy male SPF grade SD rats were randomly divided into normal control group(group A,n = 24),sham operation group(group B,n = 24),model preparation group(72).After the model preparation,the rats were randomly divided into model group(group C,n = 24),electroacupuncture group(group D,n = 24),drug group(group E,n = 24).Group A,group B,group C,group D,group E,24 rats were randomly divided into four subgroups(1D,3D,7D,14 D,6 rats / subgroup).Group A did not do any treatment;group B only separated the right common carotid artery(CCA),external carotid artery(ECA),internal carotid artery(ICA),Group C,D and E were used to establish MCAO rat model.Six hours after the successful preparation of the model,group A,group B and group C were not given any treatment,group D was given electroacupuncture(once a day),group E was given mosapride citrate solution by gavage(0.158g/ml,once in the morning and eight in the evening).All subgroups were at 1D,3D,7D,14 D after operation with four different time points,the blood and duodenal tissues of abdominal aorta were collected,the general appearance of intestinal tract was observed,and the intestinal propulsive rate was measured;the concentrations of ghrelin and VIP in serum of abdominal aorta were measured by enzyme-linked immunosorbent assay(ELISA);HE staining was used to observe the pathological changes of the duodenum of rats in each group at 1D,3D,7d and 14 d after operation.The expression of 5-HT2 a m RNA and Reg3 a m RNA in 1D,3D,7D and 14 D duodenum was detected by RT-PCR,and the expression of 5-HT2 a and Reg3 a protein in 1D,3D,7D and 14 D duodenum was detected by immunohistochemistry IHC.Results:(1)Observation on the appearance of intestinal body of rats in each group: the intestines of rats in group A and group B were arranged orderly at each time point,and there was no difference in the diameter of the intestines.The appearance color of the intestines was light red,and the surface was smooth,without swelling or atrophy.Compared with group A,the intestines of rats in group C were adhered to each other at each time point,and the diameters of the intestines were different.Some of the intestines were atrophy or swelling,and the appearance color became dark red or white.In group D,E compared with group C,intestinal adhesions were reduced,the diameter of the intestine was basically the same,sometimes the intestine atrophied,and the appearance color was light red and white.(2)HE pathological observation of duodenal tissue in each group:there was no pathological change in duodenal tissue of group A and group B,and the structure of each layer of intestinal tissue was intact and undamaged;the villi of intestine were slender and arranged orderly;the epithelial cells of intestine were intact and columnar,with obvious goblet cells and central chylous duct;there was no inflammatory cell infiltration,hyperemia or hemorrhage.In group C,the lesions of intestinal tissue were the most serious,the injury of intestinal tissue was obvious,the morphological changes of intestinal villi were lodging and atrophic,the epithelial cells of intestine were seriously exfoliated,the pan’s cells in the lower part of intestinal gland were decreased,the infiltration of inflammatory cells was serious,and the morphology of small intestinal crypt was lost.In group C,the damage of 3D group was the most serious,and the pathological changes gradually decreased with the extension of time.At the 14 th day,the normal group water was still not recovered Flat.D.There were different degrees of pathological changes in the tissues of group E,among which there was no significant difference between the three-dimensional group and group C in the transverse direction;with the increase of time,the pathological changes gradually reduced,and at the 14 th day,group D was close to the level of group A;compared with group C at the same time,the pathological changes of group D and group E were lighter,but the damage was more obvious than group B.(3)Comparison of small intestine propulsion rate of rats in each group: compared with group A at the same time point,the small intestine propulsion rate of group C,group D and group E decreased significantly in 1D,3D,7d and 14 d after operation(P < 0.05);the small intestine propulsion rate of group C increased gradually in 7d and 14 d after operation(P < 0.05),but failed to return to the normal level;compared with group C at the same time point,the small intestine propulsion rate of group D,group E increased significantly in 3D,7d and 14 d after operation Compared with 1D in group D and group E,there was a significant difference in the rate of small intestine propulsion between group D and group E(P < 0.05).(4)Comparison of serum ghrelin concentration of rats in each group: compared with group A,the serum ghrelin concentration of rats in each group decreased(P < 0.05).With the prolongation of time,the serum concentration of ghrelin in group B,C,D and e increased significantly(P < 0.05).Compared with group A,group C did not recover to the same level 14 days after operation.Compared with 1D in group D and group E,there was a significant difference in the level of serum ghrelin(P <0.05)at 7d and 14 d after operation,and the level of serum ghrelin was close to that of normal group at 14 d after operation.(5)Comparison of serum VIP concentration of rats in each group: compared with group A,the serum VIP concentration of rats in each group decreased significantly(P < 0.05).Compared with 1D group,the concentration of VIP in group C,D and e increased significantly(P < 0.05).Group C failed to return to the normal level within 14 days after operation.There was no significant difference in 1D between group A and group B(P > 0.05).There was no significant difference among groups a,B,C and D(P > 0.05).(6)Comparison of the relative expression of 5-HT2 A m RNA in the duodenum of rats in each group: compared with group A at the same time point,the relative expression of 5-HT2 A m RNA in group C,group D and group E increased significantly at postoperative 1D(P < 0.05),in which group C increased significantly compared with group D and Group E(P < 0.05);compared with group C at the same time point,the relative expression of 5-HT2 A m RNA in the duodenum of group D and group E increased significantly at postoperative 3D,7d and 14d(P < 0.05)There was significant difference in the relative expression of 5-HT2 A m RNA between group C and 1D(P < 0.05);there was significant difference in the relative expression of 5-HT2 A m RNA between group C and 1D(P < 0.05);there was significant difference in the relative expression of 5-HT2 A m RNA between group D and E and 1D(P < 0.05).(7)Comparison of the relative expression of reg3 a m RNA in the duodenum of rats in each group: compared with group A at the same time point,the relative expression of reg3 a m RNA in the duodenum of group C,D and e decreased significantly at 3D,7D and 14 D after operation(P < 0.05);compared with group C at the same time point,the relative expression of reg3 a m RNA in the duodenum of group D and e decreased significantly at 3D,7D and 14 D after operation(P < 0.05)There was a significant difference in the relative expression of reg3 a m RNA between group C and 1D after operation(P < 0.05);there was a significant difference in the relative expression of reg3 a m RNA between group C and 1D after operation(P < 0.05);there was a decrease in the relative expression of reg3 a m RNA between group D and E and 1D after operation(P < 0.05).(8)The expression level of 5-HT2 A protein in duodenum of rats in each group: compared with group A at the same time point,the expression level of 5-HT2 A protein in group C,group D and group E increased significantly(P < 0.05).Compared with group C at the same time point,the expression of group D and group E decreased significantly at 3D,7D and 14 D after operation(P < 0.05).Compared with 1D group,the expression of5-HT2 A protein in 3D,7D and 14 D groups increased significantly(P < 0.05).(9)Comparison of the expression level of reg3 a protein in the duodenum of rats in each group: compared with group A at the same time point,the expression level of reg3 a protein decreased significantly in group C,D and e at 3D,7D and 14 D after operation(P < 0.05).Compared with group C at the same time point,the expression of group D and group E increased significantly at 3D,7D and 14D(P< 0.05).Compared with 1D group,the expression of reg3 a protein in 3D,7D and14 D group decreased significantly(P < 0.05).Conclusion:(1)In the acute stage of MCAO rats,the intestinal function is abnormal,the expression of 5-HT2 a in duodenum tissue is increased,and the expression of reg3 a is decreased;(2)Electroacupuncture at Zusanli and Neiguan Points can improve the intestinal function of MCAO rats,the mechanism may be related to the expression of 5-HT2 a and reg3 a. |
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