Effect Of Leonurine On The Reproductive Performance Of Mice With Multiple Superovulation

Multiple superovulation is a key technology in livestock production,embryo engineering and in vitro embryo production.But it could cause oxidative stress to animal ovaries and early embryos,which affects animal reproductive function.Leonurine(LEO)has the ability to scavenge superoxide.And it can regulate the balance between reactive oxygen species and antioxidant defense system to reduce oxidative stress.In this study,mice are used as the model to analyze the effects of LEO on the early embryonic development potential and antioxidant capacity of multiplesuperovulation mice.At the same time,we counted the litter size of the multiplesuperovulation mice treated with LEO,and discussed the effect of LEO on the reproductive potential of the multiple-superovulation mice.1.The effects of multiple superovulations on ovarian function and early embryonic development potential in mice were investigated.We compared the ROS and GSH differences among early embryos of mice from different groups by observing the ovarian antioxidant capacity of mice with one,three,four and five times of superovulation(R1,R3,R4 and R5),and calculated the oocyte harvest number,fertilization rate,damage rate,early embryo division rate and blastocyst rate at the same time.Through the above data analysis,we further explored the effect of multiple superovulation on the early embryonic development potential of mice,and screened out the multiple superovulation group with significantly decreased ovarian function and early embryonic development potential for subsequent experiments.The results showed that,with the increase in the number of superovulation,the ovarian antioxidant capacity of mice decreased.And the quality of oocytes decreased,the level of ROS in early embryos increased,and the development potential decreased.In particular,when the number of ovulation reached 3(R3 group),compared with one ovulation(R1 group),the ovarian antioxidant capacity of R3 group significantly decreased(P < 0.05)and the embryo blastocyst development rate significantly decreased(29.39 ± 3.97 vs.74.65 ±7.93,P < 0.05),but the early embryo ROS level was significantly higher than that of R1 group(54.08 ± 5.54 vs.17.59±2.86,P < 0.05).Therefore,three times superovulation(R3)was selected as a follow-up test treatment.2.The effect of LEO on the reproductive potential of mice with multiple superovulation was investigated.In order to screen the optimal administration of LEO to the R3+LEO group,we compared the effects of LEO on oocyte quality and its early embryonic developmental potential by simultaneous gavage of mice at different concentrations(0,2,4,6,8 and 10 mg/(kg.d))during three superovulations.Then we analyzed the relationship between the ovarian antioxidant capacity of mice in the R1,R3 and R3+LEO groups,the intra-embryonic GSH content in early embryos,the ROS levels,mitochondrial function,embryonic development rate,total blastocyst cell number,the reproductive performance of mothers and growth performance of newborn mice.Based on the above data,we further explored the effect of LEO on the reproductive potential of multiple-superovulation mice.The result showed that gavage of 8 mg/(kg.d)LEO improved the ovarian antioxidant capacity,oocyte quality and early embryonic development potential in the thrice superovulated mice.Among them,the embryonic blastocyst development rate was significantly higher in the R3+LEO group compared with the R3 group(63.82±5.65 vs.26.64±7.93,P < 0.05),and the early embryonic GSH content(14.27±0.17 vs.11.30±0.31,P < 0.05)and JC-1 polarity(1.89±0.08 vs.1.13±0.18,P < 0.05)were also significantly higher,while ROS levels were significantly lower(37.68 ± 1.50 vs.54.08 ± 5.34,P < 0.05).In addition,the reproductive performance of females and growth performance of newborn mice in the R3+LEO group were significantly higher than those in the R3 group(P < 0.05),showing that the optimal dose of LEO administration was 8 mg/(kg.d).The result displayed that gavage of appropriate amount of LEO during multiple superovulation could improve the reproductive potential of multiple superovulated mice.In conclusion,multiple superovulation caused oxidative stress in mouse ovaries and early embryos,affecting the reproductive function of mice.The gavage of 8mg/(kg.d)of LEO during multiple superovulation could improve the antioxidant capacity of mouse ovaries,oocyte quality and embryonic development.It also can reduce oxidative stress in early embryos and protect mitochondrial function.Finally,all of those increase the reproductive potential of mice with multiple superovulation.