Short-term stromal alterations in the rat ventral prostate following alloxan-induced diabetes and the influence of insulin replacement

Abstract

The stromal microenvironment is pivotal to prostate physiology and malign transformation. Diabetes leads to testosterone withdrawal and affects the prostate stromal compartment and smooth muscle cells in a similar way to that observed after castration. However the response of these cells and their involvement in extracellular matrix remodeling is not satisfactorily understood. We investigated the changes caused in the short term (one week) by alloxan-induced diabetes in the stromal components of the rat ventral prostate (VP) with an emphasis on morphological alterations of stromal cells, their conversion to a myofibroblast phenotype and the remodeling of extracellular matrix and the influence of insulin therapy. Adult male Wistar rats were assigned into untreated diabetic (n=12), insulin-treated (n=8) diabetic and control (n=10) groups. Diabetes was induced by means of the injection of alloxan (40 mg/kg b.w.), while the control animals received saline solution only. Insulin (5 UI) was administered daily for one week after diabetes diagnosis. Testosterone and estrogen plasma levels were determined. VP was analyzed using transmission electron microscopy. The main stromal cells were identified by means of light microscopy, using immunocytochemistry for specific markers - vimentin for fibroblasts, α-actin for smooth muscle cells (smc) and vimentin/calponin for myofibroblasts, following the estimation of their relative frequency and absolute volume by means of stereology. After one week diabetes led to a marked decrease in testosterone levels and an atrophy of about 35% in the VP. The relative frequency of smc and collagen fibers increased in the VP of diabetic rats but their absolute weight remained unchanged. Experimental diabetes promptly altered smc morphology which assumed at the ultrastructural level a shrunken appearance with the approximation of cytoplasmic dense bodies and also exhibited a decreased immunoreactivity to calponin. The conversion of stromal cells to a myofibroblast phenotype did not occur in alloxan-induced diabetes, as evaluated by double immunoreaction to calponin and vimentin. Insulin treatment maintained testosterone levels and preserved at least partly the cell morphology and collagen fiber organization of the prostate stroma in short-term diabetes. The apparent collagen increase observed by means of microscopic analysis in the stromal prostate compartment in the short term after diabetes is mainly associated with gland atrophy and does not involve the formation of new collagen fibers, the generation of myofibroblast-like cells or the acquisition of a secretory phenotype by stromal cells.