ABSTRACT
BACKGROUND: Male reproductive tract infection and inflammation are important aetiological factors of infertility. Experimental Autoimmune Orchitis (EAO) is a model of chronic inflammation useful to study mechanisms of inflammatory reactions leading to testicular impairment. EAO is characterised by interstitial cell infiltrate of lymphomonocytes, producers of pro-inflammatory cytokines involved in germ cell apoptosis. Nitric oxide (NO), a free radical promoting immune cell activation and apoptosis, is synthesised by conversion of l-arginine to l-citrulline catalysed by NO synthase (NOS). The NOS isoforms are: constitutively endothelial (e) and neuronal (n) NOS and inducible (i) NOS. OBJECTIVES: Although the NO-NOS system was found to be up-regulated by pro-inflammatory mediators in immune and non immune testicular cells, data on its regulation in chronic inflammatory states is lacking. METHODS AND RESULTS: EAO was induced in rats by active immunisation with spermatic antigens and adjuvants; control (C) rats were injected with adjuvants. Untreated normal (N) rats were also studied. We demonstrated that iNOS, eNOS and nNOS was mainly expressed by interstitial cells in N and C rats and that in EAO NOS was up-regulated and also expressed by tubular cells. Constitutive and inducible NOS content (Western blot) as well as NO production and activity increased in the testis of rats with EAO. iNOS content and activity were selectively up-regulated in the testis of rats with orchitis. Flow cytometric analysis of NOS isoforms in testicular macrophages (M) showed that the percentage of ED1(+)ED2(-) and ED1(+)ED2(+) M subsets, expressing constitutive and iNOS isoforms was significantly higher in EAO, but no change in the percentage of ED1(-)ED2(+) resident M was observed compared to C rats. M from EAO rats also released more NO than C and N rats. CONCLUSIONS: In testis of rats with EAO, NO-NOS system was up-regulated and both testicular M and cells from seminiferous tubules contributed to NO increase. NO over production in orchitis was generated mainly by increased iNOS content and activity.
Subject(s)
Autoimmune Diseases/metabolism , Nitric Oxide Synthase/metabolism , Nitric Oxide/metabolism , Orchitis/metabolism , Testis/metabolism , Adjuvants, Immunologic , Animals , Autoimmune Diseases/chemically induced , Autoimmune Diseases/enzymology , Blotting, Western , Disease Models, Animal , Flow Cytometry , Humans , Immunohistochemistry , Macrophages/enzymology , Macrophages/metabolism , Male , Nitric Oxide Synthase Type I/metabolism , Nitric Oxide Synthase Type II/metabolism , Nitric Oxide Synthase Type III/metabolism , Orchitis/chemically induced , Orchitis/enzymology , Rats , Rats, Sprague-Dawley , Testis/enzymology , Testis/pathology , Up-RegulationABSTRACT
OBJECTIVE: To investigate the effect of immunological orchitis on spermatic specific enzyme and fertility. METHODS: Experimental allergic orchitis (EAO) model of guinea pigs was duplicated. The histological and morphological changes of spermatic acrosomal protease and hyaluronidase, lactate dehydrogenase, sperm in epididymis and testes were observed by means of enzyme kinetical spectrophotometry and gelatin fixation of substrate thin membrane. RESULTS: The activity of acrosomal protease, hyaluronidase and spermatic cytoplasmic lactic dehydrogenase in the epididymis acrosomal enzyme system became low, and so did the quality of sperm in epididymis. Remarkable morphological changes of spermatogenic cells developed in the convoluted seminiferous tubules. CONCLUSIONS: EAO remarkably affects the fertility of male guinea pigs. The orchis and epididymal sperms might be the sites of action.
Subject(s)
Fertility/physiology , Orchitis/physiopathology , Spermatozoa/enzymology , Acrosin/metabolism , Animals , Autoimmune Diseases/enzymology , Autoimmune Diseases/pathology , Autoimmune Diseases/physiopathology , Disease Models, Animal , Guinea Pigs , L-Lactate Dehydrogenase/metabolism , Male , Orchitis/enzymology , Orchitis/pathology , Spermatozoa/pathology , Testis/pathologyABSTRACT
The presence of hyaluronidase in preparations of Treponema pallidum was previously shown using acidified bovine serum albumin reactions and Ouchterlony immunodiffusion. To expand on these preliminary findings more sensitive techniques of viscometry, additional immunologic reactions, and altered capillary permeability were used to characterize treponemal-associated hyaluronidase. The pathogens T. pallidum and T. pertenue degraded hyaluronic acid, whereas the nonpathogens T. denticola and T. vincentii did not. As syphilitic infection progressed, hyaluronidase activity decreased; organisms harvested from 14-day testicular infections degraded hyaluronic acid less rapidly than organisms from 4-day infections. Uninfected rabbit testicular extract also exhibited significant enzyme activity. The neutralizing activity of immune sera was decreased by prior adsorption with bovine hyaluronidase, suggesting that some of the neutralizing factors are associated with this enzyme. Radioimmunoassay was used to quantitate antibodies to hyaluronidase in immune sera. Antihyaluronidase sera were isolated from rabbits immunized with bovine hyaluronidase. Treponema pallidum, as well as uninfected rabbit testicular extract, cross-reacted with these antisera. Immunofluorescence indicated that the hyaluronidase was uniformly distributed along the treponemal surface. As a final indicator of hyaluronidase activity, alterations in capillary permeability were detected 1 h after intradermal injection of T. pallidum.
