The effect of "Zhibian(BL 54)-to-Shuidao(ST 28)" acupuncture technique on the ex-pression of exogenous apoptotic factors in mice with poor ovarian response
Objective By observing the effect of"Zhibian(BL 54)-to-Shuidao(ST 28)"acupuncture on the expression of exoge-nous apoptotic factors in poor ovarian response(POR)mice,the improvement of acupuncture on ovarian function was discussed.Meth-ods Thirty C57BL/6 female mice with regular estrous cycle were randomly divided into blank group,model group and acupuncture group(n=10).The model group and the acupuncture group were given tripterygium glycosides suspension(50 mg·kg-1)by gavage for 14 days,and the blank group was given an equal volume of 0.9％sodium chloride solution for 14 days,once a day for 20 min.After the modeling was completed,the acupuncture group was treated with"Zhibian(BL 54)-to-Shuidao(ST 28)"acupuncture for 14 days.The blank group was only fixed without other interventions.The three groups were treated at the same time,and the corresponding indica-tors were detected the next day.The general condition of mice in each group was observed and the body weight of mice before death was recorded;the changes of estrous cycle in mice were observed by vaginal exfoliated cell smear;the number of eggs,ovarian wet weight and ovarian index of mice were detected.The levels of anti-mullerian hormone(AMH),follicle-stimulating hormone(FSH),estradi-ol(E2)and luteinizing hormone(LH)in serum were detected by enzyme-linked immunosorbent assay(ELISA);Hematoxylin-eo-sin(HE)staining was used to observe the morphological structure of ovary;the apoptosis rate of positive cells in ovarian tissue was de-tected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)staining;the mRNA and protein expres-sions of Fas cell surface death receptor(Fas),FasL,fas-associated death domain(FADD),tumor necrosis factor-related apoptosis-inducing ligand(TRAIL),death receptor 5(DR5),cysteine-aspartic acid protease(Caspase)-8 and Caspase-3 in ovarian tis-sues were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction(RT-PCR)and western blot(WB),respectively.Results Compared with the blank group,the rate of estrous cycle disorder in the model group was increased(P＜0.01).The number of retrieved oocytes,ovarian wet weight and ovarian index decreased(P＜0.01).The contents of FSH and LH in serum increased(P＜0.01),and the contents of AMH and E2 decreased(P＜0.01).The number of follicles at all levels in ovarian tis-sue decreased,the number of atretic follicles increased and the pathological morphology became worse.The apoptosis rate of positive cells was increased(P＜0.01).The mRNA expression levels of Fas,FasL,FADD,TRAIL,DR5,Caspase-8 and Caspase-3 were decreased(P＜0.01).The relative expression levels of Fas,FasL,FADD,TRAIL,DR5,Caspase-8 and Caspase-3 proteins were increased(P＜0.01,P＜0.05).Compared with the model group,the disorder rate of estrous cycle in the acupuncture group was de-creased(P＜0.01);The number of retrieved oocytes,ovarian wet weight and ovarian index were increased(P＜0.01);The contents of FSH and LH in serum decreased(P＜0.01),and the contents of AMH and E2 increased(P＜0.01);The number of atretic follicles in ovarian tissue decreased,the number of follicles at all levels increased and the granulosa cells were arranged neatly;the apoptosis of pos-itive cells decreased(P＜0.01);the apoptosis rate of positive cells decreased(P＜0.01);the mRNA expression levels of Fas,FasL,FADD,TRAIL,DR5,Caspase-8 and Caspase-3 were decreased(P＜0.01);the relative expression levels of Fas,FasL,FADD,TRAIL,DR5,Caspase-8 and Caspase-3 proteins were decreased(P＜0.01,P＜0.05).Conclusion The"Zhibian(BL 54)-to-Shuidao(ST 28)"acupuncture method may improve the ovarian dysfunction of POR mice,reduce the apoptosis of ovarian granulosa cells and improve the ovarian response by regulating the key factors in the exogenous apoptotic pathway.
万方数据
