Abstract
Hyperprolactinemia‐induced hypogonadism has been linked to a dysfunction of the hypothalamus‐pituitary‐testis axis. The direct inhibitory effects of prolactin on the testicular release of testosterone have also been demonstrated, though their mechanisms remain unclear. Incubation of rat testicular interstitial cells (TICs) with prolactin stimulated the release of testosterone. TICs from rats with anterior pituitary‐grafting‐induced hyperprolactinemia release lower amounts of testosterone than controls. However, Leydig cells isolated from anterior pituitary‐grafted rats release a greater amount of testosterone. These paradoxical observations have remained unexplained. This study examined the roles of testicular interstitial macrophages and of their product, tumor necrosis factor‐α (TNF‐α), in regulating Leydig cells under condition of hyperprolactinemia. Hyperprolactinemia was induced by grafting two anterior pituitary glands of rats under the renal capsule. Control animals were grafted with rat cortex tissue. The rats were sacrificed 6 weeks later. TICs and macrophages, and Leydig cells were isolated for in vitro incubation and drugs challenge. Testosterone released by testicular interstitial or Leydig cells was measured by radioimmunoassay. TNF‐α concentration in the medium of TICs or macrophages was measured by enzyme‐linked immunosorbent assay (ELISA). A dose‐dependent stimulation of TNF‐α secretion in the medium of TICs or macrophages by the prolactin challenge was observed. Higher amounts of TNF‐α were released by TICs in the anterior pituitary‐grafted rats than in the control group. In contrast, the release of TNF‐α by testicular interstitial macrophages isolated from the anterior pituitary‐ and cortex‐grafted groups was quantitatively similar. Challenge with human chorionic gonadotropin did not modify the TNF‐α release by testicular interstitial macrophages in either group. Challenge of Leydig cells with TNF‐α inhibited their release of testosterone stimulated by human chorionic gonadotropin, but not their basal testosterone release. These different patterns of testosterone release in TICs versus Leydig cells cultures in anterior pituitary‐grafted rats may be due to the influence of testicular interstitial macrophages. These observations correlate with in vivo conditions, where prolactin increases the release of TNF‐α by testicular interstitial macrophages, which, in turn, decreases the human chorionic gonadotropin‐stimulated release of testosterone by Leydig cells. In summary, hyperprolactinemia‐induced hypogonadism involves a mechanism of prolactin‐originated, macrophage‐mediated inhibitory regulation of testosterone release by Leydig cells. TNF‐α, one of the cytokines secreted by macrophages, may play a key role in this mechanism. © 2002 Wiley‐Liss, Inc.