Abstract
STUDY QUESTIONWhat are the consequences of polycystic ovary syndrome (PCOS) pathology and metformin-pretreatment in vivo in women with PCOS on the metabolism and steroid production of follicular phenotype- and long-term cultured-granulosa cells (GC)?SUMMARY ANSWERPCOS pathology significantly compromised glucose metabolism and the progesterone synthetic capacity of follicular- and long-term cultured-GCs and the metabolic impact of PCOS on GC function was alleviated by metformin-pretreatment in vivo.WHAT IS KNOWN ALREADYGranulosa cells from women with PCOS have been shown to have an impaired insulin-stimulated glucose uptake and lactate production in vitro. However, these results were obtained by placing GCs in unphysiological conditions in culture medium containing high glucose and insulin concentrations. Moreover, existing data on insulin-responsive steroid production in vitro by PCOS GCs vary.STUDY DESIGN, SIZE AND DURATIONCase-control experimental research comparing glucose uptake, pyruvate and lactate production and progesterone production in vitro by GCs from three aetiological groups, all undergoing IVF; healthy control women (Control, n = 12), women with PCOS treated with metformin in vivo (Metformin, n = 8) and women with PCOS not exposed to metformin (PCOS, n = 8). The study was conducted over a period of 3 years between 2007 and 2010.PARTICIPANTS/MATERIALS, SETTING, METHODSRotterdam criteria were used for the diagnosis of PCOS; all subjects were matched for age, BMI and baseline FSH. Individual patient cultures were undertaken with cells incubated in a validated, physiological, serum-free culture medium containing doses of 0–6 mM glucose and 0–100 ng/ml insulin for 6 h and 144 h to quantify the impact of treatments on acute and long-term metabolism, respectively, and progesterone production. The metabolite content of spent media was measured using spectrophotometric plate reader assay. The progesterone content of spent media was measured by enzyme-linked immunosorbent assay. Viable GC number was quantified after 144 h of culture by the vital dye Neutral Red uptake assay.MAIN RESULTS AND THE ROLE OF CHANCEGranulosa cells from women with PCOS pathology revealed reduced pyruvate production and preferential lactate production in addition to their reduced glucose uptake during cultures (P < 0.05). Metformin pretreatment alleviated this metabolic lesion (P < 0.05) and enhanced cell proliferation in vitro (P < 0.05), but cells retained a significantly reduced capacity for progesterone synthesis compared with controls (P < 0.05).LIMITATIONS, REASONS FOR CAUTIONAlthough significant treatment effects were detected in this small cohort, further studies are required to underpin the molecular mechanisms of the effect of metformin on GCs.WIDER IMPLICATIONS OF THE FINDINGSThe individual patient culture strategy combined with multifactorial experimental design strengthens the biological interpretation of the data. Collectively, these results support the notion that there is an inherent impairment in progesterone biosynthetic capacity of the GCs from women with PCOS. The positive, acute metabolic effect and the negative long-term steroidogenic effect on GCs following metformin exposure in vivo may have important implications for follicular development and luteinized GC function when the drug is used in clinical practice.STUDY FUNDING/COMPETING INTEREST(S)No competing interests. This work was supported by the UK Medical Research Council Grant Reference number G0800250.
Highlights
Polycystic ovary syndrome (PCOS) is a condition of primary ovulatory dysfunction associated with metabolic disturbances (Wild et al, 2000; Venkatesan et al, 2001; Wijeyaratne et al, 2002; ESHRE/ASRM, 2004)
The study population contained three aetiological subgroups, namely, women with normal ovaries (Control n 1⁄4 12), women with PCOS not treated with metformin during their IVF/ICSI treatment (PCOS n 1⁄4 8) and women with PCOS treated with metformin during their IVF/ICSI treatment (Metformin n 1⁄4 8)
When Wu et al (2003) compared the effects of insulin and IGF-1 on human GC glucose metabolism, the results showed that the affinity and effect of IGF-1 was much more potent than that of insulin on GC glucose uptake in women with PCOS
Summary
Polycystic ovary syndrome (PCOS) is a condition of primary ovulatory dysfunction associated with metabolic disturbances (Wild et al, 2000; Venkatesan et al, 2001; Wijeyaratne et al, 2002; ESHRE/ASRM, 2004). Glucose, metabolized under anoxic conditions to lactate, was shown to be the preferred energy substrate required to support the gonadotrophin-induced differentiation of sheep GCs in vitro; fructose and pyruvate, but not galactose, were consumed as alternative sources of energy. Purcell et al has demonstrated blunted insulin-stimulated glucose uptake in vitro by cumulus cells from women with PCOS (Purcell et al, 2012). Previous studies which have cultured GC over 48 h in vitro have demonstrated that insulin-induced glucose uptake by GCs in response to increasing doses of insulin is much lower in GCs from anovulatory patients with PCOS than from women with either normal ovaries or ovulatory polycystic ovaries (Rice et al, 2005). Caution needs to be exercised as many of these results were obtained by placing the cells in highly unphysiological conditions in serum-containing medium with high glucose and insulin concentrations
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