Chlorogenic acid supplementation during in vitro maturation improves maturation, fertilization and developmental competence of porcine oocytes

Abstract

Chlorogenic acid (CGA) is a quinic acid conjugate of caffeic acid, and a phytochemical found in many fruits and beverages that acts as an antioxidant. The present study investigated the effects of CGA supplementation during in vitro maturation (IVM), on in vitro development of porcine oocytes, to improve the porcine in vitro production (IVP) system. Oocytes were matured either without (control) or with CGA (10, 50, 100 and 200 μM). Subsequently, the matured oocytes were fertilized and cultured in vitro for 7 day. The rates of maturation, fertilization and blastocyst formation of oocytes matured with 50 μM CGA were significantly (p < .05) higher than those of the control oocytes. Hydrogen peroxide (H O ) is one of the reactive oxygen species and induces DNA damage in porcine oocytes. When oocytes were matured with 1 mM H O to assess the protective effect of CGA, 50 μM CGA supplementation improved the maturation rate and the proportion of DNA-fragmented nuclei in oocytes compared with control oocytes matured without CGA. Moreover, when oocytes were matured with either 50 μM CGA (control) or caffeic acid (10, 50 and 100 μM), the rates of maturation, fertilization and the blastocyst formation of oocytes matured with 50 μM CGA were similar to those of oocytes matured with 10 and 50 μM caffeic acid. Our results suggest that CGA has comparable effects to caffeic acid, and IVM with 50 μM CGA is particularly beneficial to IVP of porcine embryos and protects oocytes from DNA damage induced by oxidative stress. Supplementation of CGA to the maturation medium has a potential to improve porcine IVP system.

Presence of chlorogenic acid during in vitro maturation protects porcine oocytes from the negative effects of heat stress

“…Experiment 2 demonstrated that when porcine oocytes were matured at 38.5°C, supplementation of the IVM medium with 50 μM CGA significantly improved the potential of oocytes to develop to blastocyst‐stage embryos after parthenogenetic activation, thus confirming our previous results for IVF‐derived and electroporated porcine embryos (Nguyen et al, , ). Nevertheless, when applied at 38.5°C, CGA had no effect on the quality of the resulting blastocysts, as assessed in terms of total cell number.…”

“…TA B L E 3 Effects of CGA on maturation and apoptosis rates of porcine oocytes cultured in medium lacking hypotaurine, L-cysteine, and eGF * known to exert detrimental effects on oocytes through increased intracellular accumulation of ROS, leading to oxidative stress (Agarwal et al, 2012;Matsuzuka et al, 2005;Sakatani et al, 2008). Previously we have demonstrated that CGA exerts antioxidant effects on porcine oocytes during exposure to hydrogen peroxide (Nguyen et al, 2017). In Experiment 3, depletion of all potential antioxidants from the IVM medium significantly reduced the oocyte maturation rate even at 38.5°C, which in turn was prevented by addition of 50 μM CGA to the medium.…”

“…Chlorogenic acid (CGA) is a phenolic compound found mainly in coffee and tea, as well as in many fruits and vegetables (Gonthier et al, ; Mahmood, Anwar, Abbas, & Saari, ). Previously, it has been demonstrated that CGA has ROS‐scavenging, antioxidant, and antiapoptotic activity (Wu, Lin, & Zhang, ), as well as acting as a defense factor to protect porcine oocytes from oxidative stress induced by hydrogen peroxide and electroporation conditions (Nguyen et al, , ). Furthermore, CGA has been reported to have low cytotoxic effects on cellular proliferation when the cells were treated with different concentrations of CGA (Jin et al, ).…”

Hypothermic storage of porcine zygotes in serum supplemented with chlorogenic acid

“…In our previous study, we reported that CGA is an effective antioxidant that improves the developmental competence of porcine oocytes and protects oocytes from DNA fragmentation caused by H 2 O 2 exposure (Nguyen et al, ). In the current study, similarly, we found that supplementation of the storage medium with 50 µM CGA significantly improved the rates of blastocyst formation of zygotes after storage at 25°C for 24 hr.…”

Use of natural antioxidants in in vitro mammalian embryo production

“…For more specific methods, quantification of apoptotic cells using the terminal deoxynucleotidyl transferase dUTP Nick end labeling assay (TUNEL assay) is used widely because ROS can cause DNA damage (NGUYEN et al, 2017). Another employed technique is the analysis of gene expression, especially of enzymes involved in the cellular antioxidant system such as catalase, superoxide dismutase, and glutathione peroxidase, and genes related to apoptotic pathways (ESHTIYAGHI et al, 2016;MESALAM et al, 2017).…”