Distribution Of The Activated Acupoints And Expression In The Local Activated Acupoints About CGRP And Mast Cell In Acute Intestinal Mucosal Injury Rat Model

AIM:To observe the distribution of the extravasated Evans Blue (EB) dye points at the skin and the expression in the local activated points about Calcitonin gene-related peptide(CGRP) and changes of mast cells (MCs) number and morphology after acute intestinal mucosal injury(AIMI).Methodology:(1)36adult male SD rats were randomly divided eaqually in6groups according to different concentrations of mustard oil, which are5%、7.5%、10%、12.5%、15%、17.5%, each group of6. AIMI model was duplicated by giving mustard oil through anus. After AIMI modeling observed the activity of rats,4h after AIMI modeling, the rats were treated by skin preparation on the whole body, tail-intravenous injection of EB dye (5mg/100g,50mg/ml in normal saline), The plasma extravasated EB dye points at the skin of the whole body were observed in5h,10h,15h,20h,25h respectively after AIMI modeling. Use the skin positioning combined with clustering analysis all factors and contour analysis method, analyzing the time-dependent regularity of distribution of surface skin extravasation point on the basis of internal different degree damage cases.(2)30adult male SD rats were randomly divided into normal saline control(n=15) and AIMI groups(n=15). AIMI model was duplicated by giving15%mustard oil through anus,4h after AIMI modeling, the rats were treated by skin preparation on the back, tail-intravenous injection of EB dye.5h after AIMI modeling, the skin and subcutaneous tissues of extravasated EB points (BL23or BL25region) to the extravasated EB points in the model group and the corresponding points in the normal control group were sampled, record each tissue if there has a blue point, which tissue has a blue point in AIMI groups was assigned to extravasated EB points group, which tissue has not in AIMI groups was assigned to non-extravasated EB points group. Section and stain by toluidine blue (for labeling MCs). The number of MCs in these samples was counted and the degranulation rate of MCs calculated. Use the immunohistochemical method to show that the distribution of local CGRP in the extravasated EB points.Results:(1)(1) Modeling:all rats survived within25h after modeling in5%,7.5%,10%,12.5%and15%mustard oil group while4rats died in17.5%mustard oil group. Each group of rats can be found in colon tissue mucosa varying degrees of swelling, inflammation and ulcers at the observation of microscope.(2) The EB extravasation points surface distribution: the regularity of distribution of EB extravasation points are manifested mainly in the lumbosacral spine in different concentration and different time.(3) EB extravasation points change with concentration on the distribution of the body:the parallel outline inspection:EB extravasation points behave consistently (P>0.05); Outline overlap inspection:The degree of EB extravasation point is the same (P>0.05); the horizontal outline inspection:EB extravasation points are associated with different concentrations(P<0.05), the most obvious is in the concentration of15%, followed by12.5%、7.5%、10%、5%.(4) EB extravasation points change over time on the distribution of the body:the parallel outline inspection:EB extravasation points behave consistently (P>0.05); Outline overlap inspection:The degree of EB extravasation points are the same (P>0.05); the horizontal outline inspection:EB extravasation points have no obvious change in time with EB seep completely in postoperative5h (P>0.05).(5) EB extravasation points gathered mast cell number and degranulation changes:the total number of mast cells have no statistically significant in the EB extravasation points group and the non-EB extravasation point group and normal saline control group (P>0.05); the number of mast cells degranulation are significantly more in the EB extravasation points group and the non-EB extravasation point group and normal saline control groupand normal saline group(P<0.05); the degranulation rate of MCs is also significantly higher in the EB extravasation points group and the non-EB extravasation point group and normal saline control groupand normal saline group(P<0.05).(6) EB extravasation points in CGRP expression:the expression of CGRP protein is seen in EB extravasation points group with levels higher than the non-EB extravasation points group and normal saline group (P<0.05).Conclusion:(1) AIMI promotes the plasma extravasation of EB and the plasma extravasation of EB points are mainly appear in the lumbosacral spine in different concentration and different time.(2) The mast cells are involved in the process of AIMI-induced activation of points, may be correlated with the CGRP expression level.