Eye For Diarrhea-predominant Irritable Bowel Syndrome Model Rat Colon Vip And Its Receptor 1 Expression

Objective:To observe the effect of eye-acupuncture therapy for Diarrhea-predominant-irritable bowel syndrome (D-IBS) model of rats'vasoactive intestinal peptide (VIP) and its receptor 1 (VIP1-R) expression, and further discuss the functional mechanism of eye-acupuncture therapy for D-IBS , so as to explore the experimental basis of eye-acupuncture therapy for D-IBS by applicating the experimental biology molecular techniques.Methods:Healthy male Wistar rats of SPF criteria, weighting 200±20g, a total of 40. Were randomly divided into 4 groups, control group, model group, eye acupuncture group, and Pinaverium amine group (referred to as the western medicine group),5 housed in each cage, a total of 8 cages. The method combined with Restraint and stress was used to copy D-IBS model, after 18 days modeling,the model was reproduced successfully.( identified in accordance with the model evaluation criteria ). Rats of control group were fed with standard diet and water, feeding 7 days without stimulating. Rats of Model group, were allowed to take standard diet and water liberally after modeling. Rats of Acupuncture group were treated with eye acupuncture therapy for 7 days, rats of western medicine group were forced feeding with 15mg/kg of Pinaverium bromide based on "equivalent dose" principle for 7 days. Specimen collection:At the end of treatment, all rats were killed, 1-2cm colon was separated 10cm from the anus,The colon was extended with the eye scissors along the long axis .The colon was washed by saline water, Then was packed with steriled aluminum foil , labeled and kept at -70℃. The rat colon VIP and VIP1-R mRNA expression were detected by using Reverse transcription polymerase chain reaction (RT-PCR) method .ELISA method was used to detect the VIP content of rat colon tissue, The immunohistochemical method was used to detect and analysis the rat colon VIP1-R protein expression.The application of statistical software SPSS10.0.Results:1. Particles of Defecation show: the control group, model group, eye acupuncture group and WM group significantly increased their feces particles (P <0.01). Discharge time of glass ball display: Model group, eye acupuncture group, western medicine group glass ball discharge time was significantly shorter compared with control group (P <0.01). Abdominal retraction reflex in rats with semi-quantitative score shows: compared with the control group, model group, eye acupuncture group, western medicine group abdominal contraction in rat colon expansion reaction semi-quantitative score were significantly higher (P <0.01). The histopathological examination of colon in rats: Colon tissue of each rat was normal, showed a simple columnar epithelial mucosa, lamina muscularis mucosa can be seen, visible outer loose submucosa muscularis. Epithelial integrity, no congestion, edema, ulcers, inflammatory cell infiltration, no interstitial edema ,no histopathological changes, indicating that D-IBS model reproduced successfully.2.Consideringβ-actin as an internal reference, compared with the control group, the colon tissue of VIP and VIP1-R mRNA expression of model group increased (P <0.01);compared with model group, VIP and VIP1 -R mRNA expression in the colon of eye acupuncture group and western medicine group decreased (P <0.01); There was no significant difference between eye acupuncture group and WM group in the VIP and VIP1-R mRNA expression of colon (P> 0.05).3.ELISA detected as follows: compared with control group, the relative content of VIP in the colon tissue of model group increased significantly (P <0.01);compared with model group, the relative content of VIP in the colon of eye acupuncture group and western medicine group decreased significantly (P <0.01); the latter two groups showed no significant difference (P> 0.05).4. Immunohistochemical assay showed that: compared with the control group, VIP and VIP1-R protein expression in the colon of model group increased significantly (P <0.01); compared with model group, VIP and VIP1- R protein expression in the colon of eye acupuncture group and western medicine group reduced significantly (P <0.01); the latter two groups showed no significant difference (P> 0.05).Conclusion:1 Defecation of D-IBS model Rats increased, discharge time of glass balls decreased, abdominal retraction reflex in rats with semi-quantitative score was 2 points, no pathological changes in rat colon.2 The relative content of VIP in colon tissue of D-IBS model rat increased, VIP mRNA and protein expression increased; Eye acupuncture therapy can decrease the relative content of VIP in the colon,also the VIP mRNA and protein expression .3 VIP1-R mRNA and protein expression In colon tissue of D-IBS model rat increased; eye acupuncture can decrease VIP1-R mRNA and protein expression of the colon tissue.