Protective Effects And Mechanisms Investigation Of Elaiophylin Or Kuntai Capsule On The Ovarian Function Of The Aging Ovary

Objective:Young patients receiving chemotherapy occasionally face infertility and premature ovarian failure(POF).The aim of the study reported here was to explore the protective effect of elaiophylin against cyclophosphamide(CTX)-induced ovarian damage in mice.Methods:Eighty-four C57BL/6 mice were divided randomly into six groups(14 per group).One group received nothing as normal control(NC),one elaiophylin(1 mg/kg/day,Ela)treated for 30 days,one CTX(120mg/kg)treated for one day,the other three groups(CTX+Ela)mice were intraperitoneal injected with CTX one day,then low(0.25mg/kg),middle(1 mg/kg)or high(4 mg/kg)dose of elaiophylin per day for another 30 days separately.At the end of treatment,nine mice of each group were sacrificed for sampling.Several parameters were used to observe the ovarian function,including ovarian weight,follicle number and serum concentrations of estradiol(E2)and progesterone(P).Follicles were categorized and counted,and ovarian histologic sections were stained for TUNEL.Western blot was used to assess the inner mechanism of injury and repair.Vaginal smears were taken daily after the final Ela treatment to determine estrous cycle for all of the remaining mice.Mating trials were initiated twenty days after the finally Ela treated and lasted for four months.Results:The ovarian function of mice exposed to CTX injection was improved after low and middle doses of Ela administration.One month after treatment,the ovarian weight,percentage of small follicles and serum concentrations of E2 and P in the CTX+Ela(low,mid,high)groups all recovered,approaching that of normal control group.Number of births per mice,mean litter size and the percent of regular estrous cycle in the low and middle dose of CTX+Ela groups increased significantly as compared to CTX group.Immunofluorescence revealed reduced TUNEL staining.Conclusion:Treatment with low and middle dose of Ela could partly protect the mice ovary from CTX-induced damage through anti apoptosis signal pathway.Background:Kuntai capsule,a traditional Chinese medicine,has been widely used for the clinical treatment of menopausal syndrome.However,its mechanisms remain poorly understood.Considering that aging ovaries are the primary cause of menopause,this study was designed to investigate the effects and mechanisms of Kuntai capsule on ovarian function in a novel mice model with accelerated aging ovaries.Materials andmethods:Eighty-five female C57BL/6 mice were chosen for this study.Twenty of the mice were separated into the normal control group(NC).The remaining sixty-five were used to establish the novel accelerated aging ovary model by superovulation and oxidative stress.After this,each five mice were chosen to evaluate ovarian reserve and function.Then the remaining sixty mice were randomly divided into four equal groups.One group was considered the model group(Mod).The other three groups were treated with low(0.4 g/kg),middle(0.8 g/kg)and high(1.6 g/kg)doses of Kuntai capsule intragastrically every day for 4 weeks.During the treatment,the body weight and fur condition of all mice were recorded.All the mice were forced to swim to record their exhaustive swimming time(EST),which measures their strength.Mice were then sacrificed for sampling.Ovarian reserve was evaluated using follicle counts and AMH expression.Ovarian function was evaluated using estrous cycle,sex hormone level and litter experiments.Ovarian follicles were categorized and counted to estimate ovarian reserve,and ovarian histologic sections were stained for TUNEL to detect apoptotic cells.The ultrastructure of ovarian cells was observed using transmission electron microscopy.Western blotting was used to measure expression of Bax,Bcl2,AMH and SOD2 protein.Results:Compared with the NC group,the Mod group clearly displayed worse fur condition and ovarian function.These situations showed some improvement after Kuntai capsule treatment.Specifically,the fur condition and the EST of the Kuntai capsule groups were superior to the fur condition and EST of the Mod group.In cases of damaged ovarian function,Kuntai capsulecan regulate the estrous cycles,increase hormone secretion and fertility and significantly decreaseatretic follicles.The transmission electron microscopy results revealed that Kuntai capsule rescued the ovarian ultrastructure of mice.TUNEL staining confirmed that the apoptotic cells were reduced after Kuntai capsule treatment.Western blotting revealed that Kuntai capsule can increase AMH,SOD2,and Bcl2 protein expression and decrease Bax expression.Conclusions:Kuntai capsule may improve damaged ovarian function and delay ovarian aging,which may be related to its antioxidant and anti-apoptosis effects.