Transforming growth factor-beta stimulates endometrial stromal apoptosis in vitro.

Abstract

Active growth and differentiation of endometrial stromal cells during the decidualization of early pregnancy are followed by programmed cell death (apoptosis) in the antimesometrial region of the decidua. The location and temporal specificity of decidual cell apoptosis and the identification of transforming growth factor-beta 1 (TGF beta 1) and TGF beta 2 protein and mRNA in the decidua suggested that members of the TGF beta family control stromal apoptosis by autocrine or paracrine mechanisms. To examine this possibility, ovariectomized rats were pretreated sequentially with estradiol (2 days), no treatment (2 days), and medroxyprogesterone acetate 48 h before death. Endometrial stromal cells were selectively isolated by enzymatic digestion and cultured for 48 h before the experiments. The addition of TGF beta 1 (10(-9)-10(-12) M) stimulated a dose-dependent increase in 200-basepair nucleosomal fragmentation of DNA, indicating increased apoptosis. TGF beta 1 and TGF beta 2 had an equivalent effect in stimulating stromal apoptosis. If an autocrine or paracrine mechanism is responsible for the control of stromal apoptosis, increased apoptosis in cultures at high cell density and stromal secretion of TGF beta would be expected. Increasing the density of stromal cell cultures increased levels of nucleosomal DNA fragmentation. To examine stromal secretion of growth factor, cell cultures were incubated with defined medium, and concentrated media were subjected to immunoblot analysis using antibodies specific for TGF beta 1 and -beta 2. Only the antibody to TGF beta 2 detected secreted growth factor. The concentration of the secreted growth factor increased with increasing density of cell culture. To determine whether stromal secretion of TGF beta 2 is responsible for the increasing nucleosomal DNA fragmentation observed with increasing cell density, cells were cultured with and without the addition of neutralizing antibodies to TGF beta s. The addition of monoclonal antibodies to TGF beta 1,2,3 or TGF beta 2,3 inhibited apoptosis. These data provide evidence that TGF beta 2 controls apoptosis in endometrial stromal cells in vitro by an autocrine/paracrine mechanism and suggest that this mechanism controls apoptosis in these cells during early pregnancy.

Enhanced Expression of Caspase-3 in Hypertrophic Scars and Keloid: Induction of Caspase-3 and Apoptosis in Keloid Fibroblasts In Vitro

“…Recently, Ladin et al (1998) showed that clinically relevant doses of corticosteroids and interferon-␥ induce an increase in apoptosis of keloid-derived fibroblasts. Recent investigators believe that one of the triggers of apoptosis is growth factor withdrawal during differentiation (Laster et al, 1988;Moulton, 1994;Oberhammer et al, 1993;Robaye et al, 1991). This is consistent with the present results that serum deprivation significantly increases caspase-3 proteolytic activity and apoptosis in KFB lines.…”

Apoptosis at the time of embryo implantation in mouse and rat

“…145 The lethal effect of TGF-b2 has been demonstrated in primary cultures of rat decidualized stromal cells which showed an increase in the incidence of cell death (internucleosomal DNA fragmentation) when exposed to the growth factor. 146 Other experiments suggest that uterine epithelial cells may be another target for the pro-apoptotic activity of TGF-b1. Administration of TGF-b1 into the uterine stroma of estradiol-sustained ovariectomized mice resulted in an increased incidence of cell death in the adjacent epithelial layer (condensation of chromatin and cytoplasm materials, positive TUNEL staining) 147 and addition of TGF-b1 to primary cultures of mouse epithelial cells triggered a similar effect.…”

Effect of Seminal Transforming Growth Factorß1 (TGFß1) and Glutathione on Apoptosis in Spermatozoa from Tunisian Infertile Men

“…In addition, statistically significant negative correlations were found between the level of DNA fragmentation and sperm motility (r=-0.646; p=0.001), and abnormal sperm morphology (r=0.434; p=0.001). However no evident correlation was found between the percentage of sperm DNA fragmentation and sperm count, and paternal age (p>0.05) [39].…”

“…The eosinnigrosin viability test was performed by dissolving 20μl eosin with 20μl of fresh sperm and then 20μl nigrosin [37][38][39].…”