Exploring The Expression And Significance Of Neuronal Makers In ENS Of Patients With Hirschsprung's Disease

Objectives:We study the distribution of BCL-2 protein,NSE,S-100 protein,NCAM by using immunohistochemical markers techniques in the control group and the ganglionic bowel segment and aganglionic segment of HD group, and compare the result with HE stain group. A quantitative assessment of bowel ENS was done by using computer software to help us to diagnose and know some of the pathogenesis of HD.Methods:1. 31 HD patients were examined, and the specimens were collected from the surgical resection colon in aganglionic bowel segments and ganglionic segments.Control group was colon-resected from 10 age-matched patients without HD. All specimens were 10% formalin fixed and paraffin embedded.2. All specimens were underwented HE stain and scanned first, then we use the immunostaining with BCL-2,NSE,S-100,NCAM. Immunostaining morphometric analysis of NCAM,S-100 was performed by image analysis program(Image-ProPlus). The positive stained nerve fiber density in the circular muscle layer was evaluated at×400 magnification by calculating the sum of the areas occupied by the positively stained nerve fibers per unit area of the circular muscle layer.Results:1. The ganglionic cells were detected in dilated segments of HD by HE or immunohistochemical staining. The array of nerve fibres in the muscle layer was changed; and the ganglionic cells were not detected in the aganglionic segments.2. BCL-2 immunohistochemical staining was strongly positive in mature or immature ganglionic cells. However, BCL-2 immunoreactivity was weak in peripheral nerve fibers; NSE,NCAM immunohistochemical staining were positive in ganglionic cells and nerve fibers; S-100 immunohistochemical staining was negative in ganglionic cells.3. The quantitive analysis shows that there is an obvious difference in the expression of NCAM,S-100 between the anglionic and aganglionic segments of HD.Conclusions:1. BCL-2 immunohistochemical staining have an advantage in displaying ganglionic cells. It shows better staining effect than NSE,S-100,especially in revealing immature ganglionic cells, and it is important to identify HD and HAD.2. NCAM immunohistochemical staining was observed in the cell membranes of ganglionic cells and nerve fibers. So the technique staining could be helpful to diagnosis HD combined with kytoplasm staining of BCL-2.3. The BCL-2 expression decreased in the aganglionic bowel segment. BCL-2 could play a role in the pathogenesy of HD.4. NCAM low expression may be caused by decreased nerve fibers in the circular muscle layer of aganglionic segments of HD.5. Image analysis of immunohistochemical staining may help us to observe the morphological pathological changes of HD, and it is useful for pathology study, clinical diagnosis and differential diagnosis of HD.