211 Pathological Analysis of Spermatic Dysfunction in Testicular Ischemia-Reperfusion Injury

ABSTRACT Introduction & Objectives Testicular ischemia represented by torsion needs emergency surgery to reperfuse testicular blood flow. However, spermatic dysfunction is often led despite appropriate treatment. Pathology of spermatic dysfunction following testicular ischemia-reperfusion injury (IRI) is still unclear. In previous research, Inflammation and excessive oxidative stress were thought to be involved in tissue dysfunction following IRI. We hypothesized that inflammation and excessive oxidative stress plays key role in spermatic dysfunction following testicular IRI. We investigated relevance between Inflammation / excessive oxidative stress and spermatic dysfunction using testicular IRI model mice. Materials & Methods Ten to fifteen weeks old, C57BL/6 male were used through the study. Unilateral (left side) testicular vessels were clamped and de-clamped 1 hour later. Testicular blood flow was resumed. Sham operation mice were set as control. Bilateral testes were removed at 0(during ischemia), 1, 3, 5 weeks after modeling testicular IRI mice. Inflammation and oxidative stress changes in bilateral testis were investigated using histological, biochemical and immunohistochemical analysis. Spermatic changes extracted from the epididymal cauda were investigated using computer aided sperm analysis (CASA). Results Histological analysis using HE and Masson-Trichrome (MT) stain showed invasion of inflammatory cells and destruction of tissue structure from week 1. Inflammatory analysis using comprehensive protein assay and immunofluorescent stain showed increased expression of cytokines (IL-2, IL-3, IL-17A, IL-23), chemokines (CCL2, CCL5, CXCL1, CX3CL1), testes were showed from week1. Oxidative stress and cell death analysis using representative candidates (8-OHdG, ROS) showed increased expression of representative candidates from week0. Spermatic analysis showed that obvious decreased spermatic concentration and motility were showed from week 1. In contralateral testes, slow decreased spermatic concentration and motility was showed from week 1. Conclusions Spermatic dysfunction following testicular-ischemia reperfusion injury was induced in inflammatory and excessive oxidative stress changes. Inflammatory and excessive oxidative stress changes might be a novel regulatory factor for spermatic dysfunction following testicular ischemia-reperfusion injury. Disclosure Work supported by industry: no.