Protein expression of PKCZ (Protein Kinase C Zeta), Munc18c, and Syntaxin-4 in the insulin pathway in endometria of patients with polycystic ovary syndrome (PCOS)

Rivero, Rodrigo; Garin, Claire-Alix; Ormazábal, Paulina; Silva, Andréa; Carvajal, Rodrigo; Gabler, Fernando; Romero, Carmen; Vega, Margarita

doi:10.1186/1477-7827-10-17

Cited by 15 publications

(21 citation statements)

References 29 publications

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“…The cultures were further subjected for 24 h in serumfree medium to addition of testosterone, insulin, or testosterone plus insulin, 100 nM each. Testosterone and insulin concentrations were determined in previous studies from our laboratory [10,27]. The cultures with no hormonal stimulation were used as basal in each experiment.…”

Section: Cell Culture and Treatmentsmentioning

confidence: 99%

Endometria from Obese PCOS Women with Hyperinsulinemia Exhibit Altered Adiponectin Signaling

et al. 2015

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Hyperandrogenemia, hyperinsulinemia, and obesity affect 60-70% of patients with Polycystic Ovarian Syndrome (PCOS), who exhibit an altered endometrial insulin signaling. The aim of the study was to evaluate whether hyperandrogenism, hyperinsulinism, and obesity present in PCOS patients impair the endometrial adiponectin signaling pathway. The ex vivo study was conducted on 27 samples from lean (n=9), obese (n=9), and obese-PCOS (n=9) patients. The in vitro assays were performed in immortalized human endometrial stromal cells stimulated with testosterone, insulin, or testosterone plus insulin. Serum steroid-hormones, adiponectin, glucose, and insulin; body mass index, free androgen index, ISI-Composite, and HOMA were evaluated in the 3 groups. Ex vivo and in vitro gene expression and protein content of adiponectin, AdipoR1, AdipoR2, and APPL1 were determined. Adiponectin serum levels were decreased in obese-PCOS patients compared to lean (78%) and obese (54%) controls (p<0.05). AdipoR1 protein and gene expression were increased in obese group vs. obese-PCOS and lean groups (2-fold, p<0.05). In turn, AdipoR2 protein and mRNA content was similar between the 3 groups. APPL1 protein levels were reduced in endometria from both obese groups, compared to lean group (6-fold, p<0.05). Testosterone plus insulin stimulation of T-HESC and St-T1b leads to a reduction of adiponectin, AdipoR1, AdipoR2, and APPL1 protein content in both endometrial cell lines (p<0.05), whereas, in the presence of testosterone or insulin alone, protein levels were similar to basal. Therefore, endometrial adiponectin-signaling pathway is impaired in hyperandrogenemic and hyperinsulinemic obese-PCOS patients, corroborated in the in vitro model, which could affect endometrial function and potentially the implantation process.

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Section: Cell Culture and Treatmentsmentioning

confidence: 99%

Endometria from Obese PCOS Women with Hyperinsulinemia Exhibit Altered Adiponectin Signaling

et al. 2015

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“…qRT-PCR analysis confirmed significant upregulation of 8 out of 9 genes that were identified as components of this pathway, in the ovaries of LPS-treated animals (Table 2). Of these genes, upregulation of MAPK8/JNK1 has been previously associated with oocyte dysfunction (Sobinoff et al, 2010); PKC β has been reported to be involved in oocyte activation (Carbone and Tatone, 2009); PIK3C2A has been implicated in proliferation of ovarian theca-interstitial cells (Kwintkiewicz et al, 2006); PIK3R1 and PKCZ have been associated with the incidence of PCOS (Diamanti-Kandarakis, 2008; Kim et al, 2009; Rivero et al, 2012); PKD3 has been implicated in organogenesis (Ellwanger et al, 2008); RELA and RRAS2 have been associated with ovarian tumorigenesis (Chan et al, 1994; Niesporek et al, 2007; Fan and Richards, 2010); and finally, increased expression of TLR4 (receptor for LPS) has been demonstrated to underpin impaired follicular growth and function (Herath et al, 2007). …”

Section: Resultsmentioning

confidence: 99%

Immune regulation of ovarian development: programming by neonatal immune challenge

Sominsky¹,

Sobinoff²,

Jobling³

et al. 2013

Front. Neurosci.

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Neonatal immune challenge by administration of lipopolysaccharide (LPS) produces enduring alterations in the development and activity of neuroendocrine, immune and other physiological systems. We have recently reported that neonatal exposure to an immune challenge by administration of LPS results in altered reproductive development in the female Wistar rat. Specifically, LPS-treated animals exhibited diminished ovarian reserve and altered reproductive lifespan. In the current study, we examined the cellular mechanisms that lead to the previously documented impaired ovulation and reduced follicular pool. Rats were administered intraperitoneally either 0.05 mg/kg of LPS (Salmonella Enteritidis) or an equivalent volume of non-pyrogenic saline on postnatal days (PNDs) 3 and 5, and ovaries were obtained on PND 7. Microarray analysis revealed a significant upregulation in transcript expression (2-fold change; p < 0.05) for a substantial number of genes in the ovaries of LPS-treated animals, implicated in immune cell signaling, inflammatory responses, reproductive system development and disease. Several canonical pathways involved in immune recognition were affected by LPS treatment, such as nuclear factor-κB (NF-κB) activation and LPS-stimulated mitogen-activated protein kinase (MAPK) signaling. Quantitative Real-time PCR analysis supported the microarray results. Protein expression analysis of several components of the MAPK signaling pathway revealed a significant upregulation in the expression of Toll-like receptor 4 (TLR4) in the neonatal ovary of LPS-treated animals. These results indicate that neonatal immune challenge by administration of LPS has a direct effect on the ovary during the sensitive period of follicular formation. Given the pivotal role of inflammatory processes in the regulation of reproductive health, our findings suggest that early life immune activation via TLR signaling may have significant implications for the programming of ovarian development and fertility.

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“…The minimal dose was 100 nM that elicited impairment on insulin-induced glucose uptake and therefore was chosen for further experiments (data not shown). Thus, for current experiments, cells were treated for 24 h with 100 nM testosterone (HA), according previous results from our group [ 19 ] . Control cells were exposed to control media plus vehicle.…”

Section: Cells Cell Culture and General Methodsmentioning

confidence: 99%

“…In this respect, it should be noted that endometrial glandular epithelial cells exposed to testosterone decrease signifi cantly protein and mRNA expression of GLUT4 and IRS-1 [ 21 ] . In addition, for endometrial stromal cells cultured with high testosterone levels, a reduction in PKCzeta and Munc18c protein levels is observed [ 19 ] . However, the eff ect of androgens on glucose uptake in endometrial cells has not been explored yet.…”

mentioning

confidence: 98%

Testosterone Modulates the Expression of Molecules Linked to Insulin Action and Glucose Uptake in Endometrial Cells

et al. 2013

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Polycystic ovary syndrome (PCOS) is a common hyperandrogenic disorder associated with insulin resistance. Insulin exerts its metabolic function by activating the PI3K/Akt pathway and favoring glucose uptake. Caveolin-1 is a scaffolding protein which increases insulin receptor (IR) stability. Alternatively, activation of IR increases caveolin-1 phosphorylation on tyrosine-14. Furthermore, endometrial tissue from PCOS patients is proposed to be insulin resistant; however, the particular role of testosterone in modulating the metabolic effects of insulin remains unexplored in endometrial stromal cells. To evaluate whether androgens modulate the response to insulin, T-HESCs cells were stimulated with 100 nM testosterone for 24 h and changes in the protein levels of caveolin-1, IR, and Akt were determined by Western blotting (WB). After testosterone treatment, the consequences of acute insulin stimulation were evaluated by WB analysis of phospho-S473Akt and phospho-Y14Caveolin-1, as well as by measuring glucose incorporation analyzing 2-deoxyglucose (2-DOG) uptake. For cells pretreated with testosterone, higher IR, IRS-1, and caveolin-1 protein levels compared with control conditions were detected. However, in testosterone treated cells acute insulin stimulation did not increase phospho-S473Akt and phospho-Y14caveolin-1 levels and reduced 2-DOG uptake was observed compared to control cells. Our results suggest that testosterone may have a detrimental role on the metabolic effects of insulin in endometrial stromal cell cultures. Thus, the high androgen levels in patients with PCOS may favor insulin resistance observed in endometria from these women.

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Protein expression of PKCZ (Protein Kinase C Zeta), Munc18c, and Syntaxin-4 in the insulin pathway in endometria of patients with polycystic ovary syndrome (PCOS)

Cited by 15 publications

References 29 publications

Endometria from Obese PCOS Women with Hyperinsulinemia Exhibit Altered Adiponectin Signaling

Endometria from Obese PCOS Women with Hyperinsulinemia Exhibit Altered Adiponectin Signaling

Immune regulation of ovarian development: programming by neonatal immune challenge

Testosterone Modulates the Expression of Molecules Linked to Insulin Action and Glucose Uptake in Endometrial Cells

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