Effect And Mechanism Of Curcumin On Premature Ovarian Failure

The rate of decreased ovarian reserve(DOR)is about 10%in infertile women.Among them,the incidence of premature ovarian failure(POF)is 1-2.8%in women of childbearing age.Most dor patients are treated in reproductive centers due to infertility.POF is related to the dramatic increase of reactive oxygen species(ROS)in the ovary.ROS mediated ovarian oxidative stress affects follicular development,oocyte maturation and ovulation,reduces oocyte quality and embryo development potential.It not only greatly reduces the natural pregnancy rate and infertility,but also reduces the oocyte maturation rate,fertilization rate,blastocyst formation and implantation rate of assisted reproduction Pregnancy rate and live birth rate affect the clinical outcome of assisted reproduction.Therefore,the problem of low fertility of POF women has become a hot and difficult point in clinical reproductive medicine.It is urgent to conduct in-depth research on its molecular mechanism,intervention programs and pregnancy strategies.Curcumin(cur)is the active component of curcuma root and stem.Studies have shown that curcumin can inhibit intracellular oxidative stress through a variety of ways,so it can improve the pathological state of the disease and produce good curative effect.POF is related to the rapid increase of ROS in ovary.Oxidative stress in ovarian microenvironment is the pathophysiological mechanism of POF.So,does curcumin exert ovarian protection through the mechanism of antioxidant stress,and thus delay the occurrence and development of POF?At present,the effect and mechanism of curcumin on premature ovarian failure have not been reported.In this study,we established a POF mouse model induced by D-galactose(D-gal)to observe the effects of curcumin on ovarian protection and premature ovarian failure;Objective to investigate the mechanism of curcumin induced oxidative stress and its signal pathway in ovarian granulosa cells.This study elucidates the role and molecular mechanism of curcumin in ovarian protection and delaying premature ovarian failure,expands the application of natural antioxidant curcumin,provides theoretical basis for in vitro application of assisted reproductive technology oocyte culture,and lays a foundation for clinical application of premature ovarian failure.This study is divided into three parts:1?In this experiment,D-gal model was established by 42 days of D-gal injection in c57b1/6 mice,and curcumin treatment group was established by 42 days of curcumin(100mg/kg/day)injection in D-gal model mice.The control group was the normal mice without D-gal intervention or curcumin intervention.The results showed that the level of FSH and LH in the D-gal model was significantly increased(P<0.05 and P<0.01),the serum E2 and P levels were significantly decreased(P<0.01),and the expression of AMH mRNA in ovary decreased significantly(P<0.05).The experimental results showed that the injection of D-gal could establish a POF model.The number of follicles in curcumin treatment group was higher than that of D-gal model at different stages of follicular maturation and development.Curcumin can significantly reduce the number of TUNEL positive cells compared with D-gal model(P<0.01).Further study of the effect of curcumin on ovarian related signaling pathway showed that the p-Akt level of D-gal model was significantly lower than that of control group(P<0.01),curcumin treatment group was significantly higher than that of D-gal group(P<0.01),caspase-3 and caspase-9 levels were significantly lower than those in D-gal group(P<0.01).In addition,the expression levels of Nrf2 and HO-1 in D-gal model were significantly lower than that in the control group(P<0.01),while that of curcumin treatment group was significantly higher than that in D-gal group(P<0.05 and P<0.01).It can be proved that curcumin can reduce oxidative stress damage induced by D-gal by activating pi3k/akt and nrf2/ho-1.2?Oxidative stress is closely related to apoptosis of granulosa cells.Apoptosis of granulocyte caused by oxidative stress is one of the important reasons for the egg quality decline.In this experiment,we used mouse ovarian granular cells to study the effect of curcumin on oxidative stress of ovarian granulosa cells.The experiment was divided into model group(H2O2),intervention group(H2O2+cur),Nrf2 inhibitor group(H2O2+cur+m1385)and normal control group(DMSO).The concentration of H2O2 was 400 ?m,cur concentration was 10?m,m1385 concentration was 5 ?M.The results showed that the total cell mortality rate of H2O2 group was significantly higher than that of DMSO group(P<0.01),and the total apoptosis rate of H2O2+cur group was significantly decreased compared with that of H2O2 group,while the anti apoptosis effect of curcumin was significantly inhibited by adding curcumin and Nrf2 inhibitor(m1385).The expression levels of Nrf2,HO-1 and nqo-1 in H2O2 group were significantly lower than that in DMSO group(P<0.01),and that of cleaved caspase-3 and cleaved caspase-9 was significantly higher than that in DMSO group(P<0.01),while that of H2O2+cur group was significantly higher than that in H2O2+cur group,and that of clean caspase-3 and cleaved caspase-9 was significantly lower than that in H2O2 group(P<0.01).The expression of nrf2/ho-1 signal pathway related protein was inhibited by curcumin and m1385.The results showed that H2O2 increased the ROS level and apoptosis signal in the granular cells,while adding curcumin could inhibit the apoptosis and ROS level of the granular cells induced by H2O2.Curcumin can inhibit oxidative stress damage of H2O2 by promoting nrf2/ho-1/nqo-1 signaling pathway of granular cells,and reduce the expression of cleaved casase-3 and cleaved casase-9,while adding Nrf2 inhibitor m1385 can significantly reverse the inhibition effect of curcumin on oxidative stress.3?Oxidative stress of ovary not only affects the formation of eggs,the development of follicles,but also reduces the quality of oocytes and the development potential of early embryos.This study is to investigate the effect of curcumin on the fertilization ability and early embryo development ability of H2O2 model mice.The experiment was divided into H2O2 group,H2O2+cur group and DMSO control group,in which the concentration of H2O2 was 400 ?m and the cur concentration was 10 ?M.The results showed that the fertilization rate of H2O2 group decreased significantly(P<0.01)compared with DMSO group,and that of H2O2+cur group was significantly higher than that of H2O2 group(P<0.01).Compared with DMSO group,the two cell rate and blastocyst rate of H2O2 group decreased significantly(P<0.01),and the ratio of H2O2+cur group was significantly higher than that of H2O2 group(P<0.01).The quality of blastocyst and the number of apoptosis cells in blastocyst stage decreased significantly in H2O2 group(P<0.05 and P<0.01)compared with DMSO group,and significantly increased in H2O2+cur group(P<0.05 and P<0.01).This experiment proved that curcumin can significantly inhibit the increase of oxidative stress induced by H2O2,improve fertilization rate,second cell stage rate,blastocyst rate and inhibit apoptosis induced by oxidative stress.The results showed that curcumin can reduce oxidative stress,apoptosis and ovarian damage induced by D-gal by activating pi3k/akt and nrf2/ho-1,increase the number of follicles at all levels and delay premature ovarian failure;curcumin can inhibit the apoptosis and ROS level of ovarian granulosa cells induced by H2O2 by promoting nrf2/ho-1/nqo-1 signal pathway;in vitro fertilization medium Adding appropriate curcumin can inhibit the oxidative stress induced by H2O2,improve the fertilization and embryo development potential,increase the number of blastocyst cells and inhibit the apoptosis of blastocyst cells.