Abstract
BackgroundHigh concentration of glucose in culture medium affects the developmental process and the quality of the pre-implantation embryo. This study examined the effects of Zuogui Wan (ZGW) supplementation on early embryo development cultured in high-glucose medium.MethodsEmbryos were cultured in high-glucose medium with or without ZGW supplementation. Developmental rate and competence was evaluated by cleavage rate, blastocyst rate, and blastocyst total cell number, reactive oxygen species (ROS) level, glutathione (GSH) concentration, and metabolome were also measured to determine the effect of ZGW on embryo development at the cellular level.ResultsCompared with the vehicle group, supplementation of 0.01 % (v/v) ZGW to high-glucose medium significantly increased cleavage rate (80.1 ± 1.0 % vs 72.1 ± 1.3 %), blastocyst rate (50.5 ± 1.0 % vs 41.3 ± 1.7 %), and blastocyst total cell number (63.2 ± 2.2 vs 57.2 ± 1.6). ROS level was lower and GSH concentration in blastocysts was higher in ZGW-treated group. Metabolomic analysis found that the ratio of glucose to succinic acid and glucose to fumaric acid were lower in the ZGW-treated group .ConclusionsDevelopmental rates of zygotes in high-glucose culture medium were significantly lower than those in regular culture medium. ZGW supplementation significantly improved embryo development and quality in high-glucose medium. Supplementing ZGW in high-glucose medium also significantly increased total cell number and GSH concentration but decreased ROS level in blastocysts likely by modifying metabolic profile during embryo development. Together, these data suggest that supplementation of ZGW rescues high-glucose-induced detrimental effects on pre-implantation embryo development.
Highlights
High concentration of glucose in culture medium affects the developmental process and the quality of the pre-implantation embryo
Effects of Zuogui Wan (ZGW) on developmental competence of embryo To determine the effects of ZGW on developmental competence of embryo, embryos were cultured in highglucose medium supplemented with vehicle only, 0.01 %, 0.1 %, 1 % (v/v) ZGW for 5 days
Effects of ZGW on levels of reactive oxygen species (ROS) and GSH levels in embryos Embryos were cultured in high-glucose medium with or without ZGW for 5 days, and the levels of ROS and GSH were determined in the blastocysts
Summary
High concentration of glucose in culture medium affects the developmental process and the quality of the pre-implantation embryo. This study examined the effects of Zuogui Wan (ZGW) supplementation on early embryo development cultured in high-glucose medium. High-glucose level in the maternal environment induces early embryo loss, growth retardation and increases the percentage of congenital malformations, such as cardiovascular anomalies and neural tube defects [5,6,7]. High-glucose concentration exert its detrimental effect on embryo development and survival by inducing DNA damage. High level of glucose increases oxidative stress by producing large amount of ROS, which can inhibit embryo development [7, 10, 11]. High-glucose level triggers DNA damage response through oxidative stress, results in abnormal development of embryo [12].
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