Direct effect of pholyphenol‐rich plants, rooibos and ginkgo, on porcine ovarian cell functions

Abstract

The polyphenol-rich plants rooibos and ginkgo are widely used in folk medicine and in preparation of foods and drinks, but their effect on reproduction has not been properly studied yet. The aim of our in vitro experiments was to examine the possible direct effect of rooibos and ginkgo on the basic ovarian cell functions-proliferation, apoptosis and release of hormones progesterone (P4) and leptin (L). Porcine ovarian granulosa cells were cultured in the presence of rooibos or ginkgo extract (0, 1, 10 and 100 μg/ml of medium). The accumulation of markers of proliferation (PCNA and cyclin B1) and apoptosis (bax) and their mRNAs was analysed using immunocytochemistry and by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Release of P4 and L was evaluated by radioimmunoassay. It was observed that rooibos or ginkgo addition was able to inhibit proliferation (down-regulates PCNA, cyclin B1 and their mRNAs), to promote apoptosis (accumulation of bax) and to suppress both L and P4 release by ovarian cells. These data suggest a direct inhibitory (anti-proliferative, pro-apoptotic and hormone-suppressing) effect of polyphenol-containing plants rooibos and ginkgo on ovarian functions. The potential anti-reproductive effect of these medical plants is to be taken into account by their consumption.

Buckwheat, rooibos, and vitex extracts can mitigate adverse effects of xylene on ovarian cells in vitro

“…Granulosa cells from the ovaries of non-cycling pubertal gilts approximately 180 days old were isolated from follicles 2-5 mm in diameter, processed, and cultured as described previously (Štochmaľová et al 2018;Tarko et al 2018;Sirotkin et al 2019). Briefly, cells collected at a concentration of 10 6 / mL were pre-cultured in sterile DMEM/F12 1:1 medium supplemented with 10% fetal bovine serum (both from BioWhittaker TM , Verviers, Belgium) and 1% antibioticantimycotic solution (Sigma, St. Louis, MO, USA) in 16well chamber slides (Nunc Inc., International, Naperville, USA, 200 μL/well) for 3 to 4 days.…”

“…each at 10 μg/mL or combinations of xylene (0, 10, 100, or 1000 ng/mL) and one of the plant additives (10 μg/mL). These doses correspond to the doses of xylene that are typically found in the environment (Bolden et al 2015;Kim et al 2015) and the effective doses of xylene (Tarko et al 2018) and plant extracts (Ohyama et al, 2003;Sirotkin et al , 2019Štochmaľová et al 2018) used in previous in vitro experiments. The plants were minced 2 min in coffee mill and dissolved in 50 μL of DMSO to create stock solutions of 1 mg/mL of plant extract.…”

Influence of green tea constituents on cultured porcine luteinized granulosa cell functions

“…Subsequently, the cells were incubated in the presence of monoclonal antibodies against either PCNA, cyclin B1, bax, and caspase 3 (all from Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA; dilution 1:500 in PBS) for 2 h at room temperature or overnight at 4 °C. The specificity and suitability of these antibodies for analysis of the corresponding antigens in cultured porcine granulosa cells were validated previously by Western immunoblotting (Mészárosová et al, 2008;Sirotkin, 2010) and RT-PCR (Kádasi et al, 2017;Štochmaľová et al, 2018). The results obtained by Western immunoblotting and RT-PCR corresponded to the results obtained by using quantitative immunocytochemistry.…”

Rooibos (Aspalathus linearis) influence on health and ovarian functions

“…The ability of rooibos to affect markers and promoters of proliferation (PCNA, cyclin B1) and apoptosis (bax) (Sirotkin, Macejková, Tarko, Fabova, Alrezaki, et al, 2020; Sirotkin, Macejková, Tarko, Fabova, Alwasel, et al, 2020; Štochmaľová et al, 2018) suggests that it could affect female reproduction through changes in ovarian cell proliferation and apoptosis and their equilibration, which can affect ovarian follicular growth, development, atresia and fecundity (Sirotkin, 2014). Nevertheless, rooibos influence on proliferation and apoptosis was not associated with significant changes in ovarian cell viability (Sirotkin, Macejková, Tarko, Fabova, Alrezaki, et al, 2020; Sirotkin, Macejková, Tarko, Fabova, Alwasel, et al, 2020).…”

“…The few available evidence demonstrates direct action of rooibos on basic ovarian cell functions. Additions of rooibos extract at doses 1, 10 and 100 μg/ml of medium were able to inhibit proliferation (down‐regulated PCNA, cyclin B1 and their mRNAs), to promote apoptosis (accumulation of bax) and to suppress release of progesterone and leptin by cultured porcine ovarian granulosa cells (Štochmaľová et al, 2018). Subsequent studies, however, did not confirm the ability of rooibos additions at dose 10 µg/mL to affect this marker of apoptosis.…”