| Objectives1 To explore the mechanism of Kuntai Capsule on fatty acid synthase?Fas?and apoptosis-related factor ligand?FasL?in ovarian tissue of rats with premature ovarian failure;2 To explore the effect of Kuntai Capsule on repairing pathological damage of ovarian function and reproductive function in rats with premature ovarian failure.MethodsEighty-four 12-week-old SD female rats were randomly divided into two groups:ovarian function group and reproductive function group.The ovarian function and reproductive function were observed.Each group was divided into blank group,model group,Kuntai capsule high,medium and low dose group and conjugated estrogen tablet group,with 7 rats in each group.Except for the blank group,rats in the other groups were fed with Tripterygium wilfordii polyglycoside tablets?75 mg/kg for 14 days?to establish the model of premature ovarian failure.At the end of the model,the model group was given 2 mL/kg normal saline,Kuntai capsule group and conjugated estrogen tablet group were given high,medium and low doses of Kuntai capsule?1.2 g,0.6 g,0.3 g/kg,36 days?and conjugated estrogen tablets?0.075 mg/kg,36 days?respectively.The ovarian function group observed the changes of behavior,estrous cycle,enzyme-linked immunosorbent assay?ELISA?to detect the changes of serum levels of sex hormone follicle stimulating hormone?FSH?,luteinizing hormone?LH?,estradiol?E2?,anti-Mullerian hormone?AMH?,histomorphology of ovary,immunohistochemistry and real-time fluorescence quantitative PCR.The contents of FAS,FasL protein and gene of apoptosis-related factor ligand in rat ovary were detected.All female rats in reproductive function group were caged with 12 SD male rats.The general situation of pregnant rats and offspring were observed and recorded.The number of offspring,fertility rate,pregnancy rate and deformity rate of offspring of female rats in each group were observed.Results1 Effects on rat behavior:All rats in blank group did not show abnormal behavior during the whole experimental process;rats in model group showed depression,decreased activity,curling,hair removal,bow back and other manifestations during the modeling period;rats in all treatment groups showed behavioral performance of rats in model group during the modeling period,but after drug treatment,Kuntai capsule had high and medium doses.The behavioral changes of rats in group A and group B gradually became normal,while those in low dose group of Kuntai Capsule had poor therapeutic effect,manifested as poor mental state,hair removal and slight improvement of bow back.2 The estrous cycle of rats in the blank group was about 4-5 days,the estrous cycle of rats in the model group was prolonged for 6-8 days,and the estrous cycle of rats in the model group and the low dose Kuntai capsule group was prolonged or disordered compared with that in the blank group,with the disorder rates of 100%and 85.71%respectively,and the difference of estrous cycle was statistically significant?P<0.05?.3Effects of Kuntai Capsule on Serum Hormone Level in Rats:Compared with blank group and conjugated estrogen tablet group,there were significant differences in serum FSH,LH,E2 and AMH concentrations in Kuntai Capsule low dose group?P<0.05?;compared with model group,there were significant differences in serum FSH,LH,E2 and AMH concentrations in other groups?P<0.05?.4 Effects on histomorphology of ovary in rats:In the blank group,follicles at all levels were active and abundant,follicles developed well,follicles structure was clear,and blood vessels were abundant;in the model group,follicles developed poorly,follicles structure was disordered,atresia follicles and pathological fibrosis were observed;compared with the blank group and the combined estrogen tablet group,Kuntai capsule had low dosage.Compared with the model group,the other groups showed better follicular development,clearer follicular structure and fewer atresia follicles.5 Effects of Kuntai Capsule on Fas and FasL Protein and Gene Expression in Rat Ovary:Compared with blank group and conjugated estrogen tablet,Fas protein and gene expression in rat ovary in low dose Kuntai Capsule group were significantly decreased,FasL protein and gene expression were significantly increased?P<0.05?;Compared with model group,Fas protein and gene expression in other groups were significantly increased,FasL protein and gene expression were significantly increased.The expression of protein and gene decreased significantly?P<0.05?.6 Influences on the general situation of pregnant rats and offspring:Except the model group rats were not pregnant,the other groups of rats were pregnant rats.In the low dose Kuntai capsule group,the pregnant rats had poor mental health,slightly slower activity and slower growth of food intake,which was significantly lower than that in the blank group and the combined estrogen tablet group on the first day of pregnancy?P<0.05?,with a significant difference?P<0.05?,and a slightly slower growth of body mass,which was significantly lower than that in the blank group and the combined estrogen tablet group on the sixth day of pregnancy?P<0.05?.The offspring have ruddy and lustrous skin,grow fast and develop normally.7 Influences on the reproductive function of female rats:Except the model group rats were not pregnant,the other groups were pregnant rats.Compared with blank group and conjugated estrogen tablet group,Kuntai capsule low dose group had lower total litter size,pregnancy rate and pregnancy rate?P<0.05?;compared with model group,except Kuntai capsule low dose group,the other groups had more total litter size,higher pregnancy rate and pregnancy rate?P<0.05?.Conclusions1 Female rats fed with Tripterygium wilfordii polyglycoside tablets?75mg/kg,14 days?showed abnormal behavioral changes,prolonged or disordered estrous cycle,serious disturbance of serum sex hormone levels and abnormal ovarian histomorphology,suggesting that the model of premature ovarian failure was successful in rats.Experimental studies have confirmed that Tripterygium wilfordii polyglycoside tablets can inhibit the ovarian and reproductive functions of female rats,and damage the ovarian and reproductive functions of rats to varying degrees.2 Kuntai Capsule can repair pathological damage of ovarian function in reproductive age rats,protect and improve ovarian function.The mechanism may be related to the regulation of serum sex hormone level by Kuntai Capsule,which has two-way regulation effect on hormone level,down-regulation of FSH and LH levels,up-regulation of E2 and AMH levels,increase of Fas protein and gene expression,and decrease of FasL protein and gene expression.In addition,Kuntai Capsule can also improve the reproductive function of rats,improve the pregnancy rate and pregnancy rate of premature ovarian failure rats,and has no obvious teratogenicity to offspring mice.3 Kuntai capsule high and medium dose group and conjugated estrogen tablet group have similar therapeutic effect,with equivalence.The low dose group of Kuntai capsule has poor therapeutic effect,which may be related to the low dose group and low concentration of Kuntai capsule.Figure28;Table10;Reference 84... |
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