Resistin is a survival factor for porcine ovarian follicular cells

Abstract

Previously, we demonstrated the expression of resistin in the porcine ovary, the regulation of its expression and its direct effect on ovarian steroidogenesis. The objective of this study was to examine the effect of resistin on cell proliferation and apoptosis in a co-culture model of porcine granulosa and theca cells. First, we analysed the effect of resistin at 1 and 10 ng/ml alone or in combination with FSHand IGF1 on ovarian cell proliferation with an alamarBlue assay and protein expression of cyclins A and B using western blot. Next, the mRNA and protein expression of selected pro-apoptotic and pro-survival regulators of cell apoptosis, caspase-9, -8 and -3 activity and DNA fragmentation using real time PCR, western blot, fluorescent assay and an ELISA kit, respectively, were analysed after resistin treatment. Furthermore, we determined the effect of resistin on the protein expression of ERK1/2, Stat and Akt kinase. Using specific inhibitors of these kinases, we also checked caspase-3 activity and protein expression. We found that resistin, at both doses, has no effect on cell proliferation. The results showed that resistin decreased pro-apoptotic genes, which was confirmed on protein expression of selected factors. We demonstrate an inhibitory effect of resistin on caspase activity and DNA fragmentation. Finally, resistin stimulated phosphorylation of the ERK1/2, Stat and Akt and kinases inhibitors reversed resistin action on caspase-3 activity and protein expression to control. All of these results showed that resistin has an inhibitory effect on porcine ovarian cell apoptosis by activation of the MAPK/ERK, JAK/Stat and Akt/PI3 kinase signalling pathways. Reproduction (2015) 150 343-355

Effect of High Fat Dietary Intake during Maternal Gestation on Offspring Ovarian Health in a Pig Model

“…High expression of BCL2 resulted in either higher ovulation rate or litter size [40], and deletion of BCL2 led to complete blockade of follicular oocyte development [41]. BAX opposes BCL2 function and is a pro-apoptotic protein [42]. Previous studies reported that BAX is expressed in both fetal and adult ovaries [43], and BAX expression is increased in atretic follicles compared to healthy ones [44].…”

Adiponectin and resistin: potential metabolic signals affecting hypothalamo-pituitary gonadal axis in females and males of different species

“…Similarly, in pig ovary, resistin decreased pro-apoptotic genes expression, caspase activity and DNA fragmentation (Rak et al 2015b). As a molecular mechanism of resistin action on cell survival, authors have proposed the activation of several signal transduction pathways such as MAPK/ ERK1/2, JAK/STAT and PI3K (Rak et al 2015b). These results show that resistin is involved in ovarian apoptosis regulation and could regulate follicular development or atresia.…”

“…In primary human granulosa cells, treatment with adiponectin at a concentration of 5 µg/ mL does not affect the IGF1-induced cell proliferation (Chabrolle et al 2009). The published in vitro data concerning ovarian cell proliferation showed some effects dependent on the resistin doses; no effect at the lower, physiological doses (1 and 10 ng/mL) in pigs (Rak et al 2015b) and rats (Maillard et al 2011), whereas stimulated effects were observed at higher doses (100; 333 and 667 ng/mL). Similarly, in vivo, resistin induced a stimulatory effect on the expression of proliferating cell nuclear antigen (PCNA) in the ovaries of seasonally monoestrous bats (Singh et al 2014).…”

“…Recently, Singh and coworkers showed that resistin injection in bats (6.5 μg/100 g body weight/day for 12 days) increased Stat-3 phosphorylation in the ovaries. Similarly, in cultured porcine ovarian cells, resistin (10 ng/mL) activated not only the phosphorylation of Stat-3 but also MAPK ERK1/2 and Akt (Rak et al 2015b). Apoptosis is critically important for the survival of multicellular organisms by getting rid of damaged or infected cells that may interfere with normal ovarian function such as oogenesis, folliculogenesis, oocyte loss/selection and atresia.…”

Maternal High-Fat Diet During Pregnancy and Lactation has Opposite Effects on Gonadal Expression of Leptin and Leptin Receptor in Rat Dams and Their Offspring

“…The main goal of the present study was to investigate the effect of maternal HF diet during pregnancy and lactation on gonadal, periovarian, and epididymal white adipose tissue (WAT) protein expression of leptin and its receptor ObRb in dams and their offspring of both sexes in Wistar rats. It is well known that leptin, like other adipokines such as resistin [9], adiponectin [10], and chemerin [11] can affect female and male reproductive functions. The first indication that leptin is involved in regulation of the reproductive system may have been the finding that leptin acts as a signal triggering puberty, supporting the hypothesis that fat accumulation enhances maturation of the reproductive tract [12].…”