| Objectives By observing the behavior,estrus cycle,levels of serum sex hormones FSH,LH,E2,AMH,ovarian tissue and ovarian GDF-9(growth differentiation factor 9),Egr-1(early growth response gene)protein and gene expression,To investigate the therapeutic effect and mechanism of Kuntai capsule on cyclophosphamide-induced ovarian reserve dysfunction in rats.Methods Twelve 12-week-old SD female rats were randomly divided into two groups,namely the middle-model group and the post-model group.The model group included blank group,cyclophosphamide model group(model group),Kuntai capsule group(high,middle,low dose),and conjugated estrogen group(Western medicine control group);the post-mold group consisted of blank group and ring Phosphoamide model group(model group),Kuntai capsule group(high,medium,low dose),combined estrogen group(Western medicine control group).10 in each group.Except for the blank group,other rats were given an intraperitoneal injection of cyclophosphamide 75 mg/kg once to establish a rat model of ovarian hypofunction,and then the rats were given drug intervention during the modeling process and after completion of the modeling.Kuntai Capsule Group In the combined estrogen group,high-,low-dose Kuntai capsules and conjugated estrogen tablets were given to the stomach,and the blank group and the model group were given equal volume distilled water.Observed changes in behavior,estrus cycle,levels of serum sex hormones AMH,FSH,LH,and E2,changes in ovarian tissue morphology,ovarian GDF-9,and Egr-1 protein expression levels in the rats of the modeling group and the post-model group.,Ovarian GDF-9,Egr-1 m RNA expression levels.Results 1 Changes in behavior of rats: In the blank group,the rats had a good mental state,a responsive response,a pure white coat,a good degree of activity,a normal eating and drinking condition,and an increase in body weight.Rats in the model group suffered from lassitude after being injected with cyclophosphamide intraperitoneally,with ecchysis,dull coats,and hair removal.Daily water intake increased compared with the blank group,and diet was not considered.Body weight was decreased.After the injection of cyclophosphamide in the abdominal cavity of each group in the model group,the spirit was slightly sluggish,the activity was reduced,the diet was reduced,slight hair removal,and the body weight was slightly reduced.With the increase of the number of days of administration of Kuntai capsule,the spirit gradually improved.,The diet returned to normal,no obvious hair removal,weight gain,Kuntai low-dose group than other groups,the hair removal phenomenon is more obvious,weight gain is not obvious.Before the administration of the model group rats,the same conditions as the model group rats appeared,hair loss and mental deterioration were found.After the administration,the spirit of the high-dose group and the combined estrogen group gradually improved,the activity returned to normal,and the diet was normal.The hair removal phenomenon has obviously improved and the body weight has gradually increased.In the low-dose group of Kuntai rats,there was still epilation,the spirit was poor,and the weight gain was not obvious.2 Rat estrus cycle: estrus cycle of the blank group rats was 4-5 days;estrus cycle of the model group rats was extended to 8-9 days;model group of Kuntai high school dose and combined with estrogen group and after modeling The estrus cycle of the Kuntai highdose group was mostly 5-6 days;the low-dose Kuntai group and the low-dose Kuntai middle-dose group had occasional estrus,but the estrus cycle was relatively low.The estrogen group was more prolonged and the estrus cycle was mostly 6-7 days.3 Rat serum hormone levels: 1)In the model group: compared with the blank group,the serum FSH in the model group was significantly higher,E2,AMH was significantly lower(P<0.01),and LH was increased(P<0.05);Compared with the model group,the FSH of the rats in each treatment group was significantly lower,E2 and AMH were significantly higher(P<0.01),and LH was decreased(P<0.05).2)After the model group:compared with the blank group,the serum FSH of the model group was significantly increased,E2,AMH was significantly lower(P<0.01),LH was increased(P<0.05);compared with the model group,each given The serum FSH in the drug group was significantly lower,E2 and AMH were significantly higher(P<0.01),and LH was decreased(P<0.05).2)Post-mold group: compared with the blank group,serum FSH in the model group was significantly increased,E2,AMH was significantly lower(P<0.01),LH was increased but not significant(P<0.05);In comparison,the serum FSH of rats in each drug group was significantly lower,E2 and AMH were significantly higher(P<0.01),and LH was decreased but not significant(P<0.05);compared with the blank group,the KT low dose group Serum FSH levels increased in rats,E2,AMH decreased,LH increased but not significant(P <0.05).4 Rat ovary tissue morphology: In the blank group,the appearance of the ovaries of the rats was pink,and the number of follicles in the cortex was normal.All the follicular morphologies,follicles,and follicular fluids were seen in the whole group;the ovary appearance in the model group was grayish in color and the number of follicles was reduced.There were more atresia follicles and interstitial fibrosis;the ovary and the blank group of the Kuntai high school and combined estrogen group and the post-implantation high-dose group of the model group performed similarly;In the dose group and in the post-model group,the ovaries of the rats in the Kuntai low-dose group showed pinkish appearance,fewer follicles,and smaller overall follicles.5 Rat ovarian protein and mRNA expression: 1)In the model group: compared with the blank group,the expression of GDF-9 and EGR-1 protein and m RNA in the model group were significantly reduced(P<0.05);compared with the model group,The expression of GDF-9 and EGR-1 protein and m RNA in the ovary of rats in all groups were significantly increased(P<0.05).Compared with the blank group,the GDF-9 and EGR-1 proteins in rats in the low-dose group of Kuntai capsules The m RNA expression decreased(P<0.05).2)Post-model group: compared with the blank group,the expression of GDF-9 and EGR-1 protein and m RNA in the ovary of model rats decreased(P<0.05).Compared with the model group,the ovary GDF-9.The expression of EGR-1 protein and m RNA was significantly increased(P<0.05).Compared with the blank group,the expression of GDF-9 and EGR-1 protein and m RNA were decreased in the low-dose group of Kuntai Capsule(P<0.05).Conclusions 1 Kuntai Capsules can improve the behavioral changes of rats with poor ovarian reserve and correct the estrus cycle of rats,suggesting that Kuntai Capsules can improve the symptoms of DOR.2 Kuntai Capsules can reduce the level of sex hormones FSH and LH in rats and increase E2 and AMH levels,suggesting that it has the effect of adjusting serum levels of DOR.3 Kuntai capsule can repair damaged ovarian tissue,improve ovarian blood supply,promote rat follicle development,reduce follicular atresia,and increase the number of mature follicles,suggesting Kuntai capsules can improve the ovarian structure of DOR.4 Kuntai Capsules can increase the expression of GDF-9 and EGR-1 protein and m RNA in rat ovary,suggesting that Kuntai Capsules can improve ovarian reserve by altering the expression of GDF-9 and EGR-1 in the ovaries.5 The treatment effect of the model group is better than that of the post-model group.It is suggested that in the clinical diagnosis and treatment,once the medicine with the gonad damage is applied,the Kuntai capsule should be applied as soon as possible to better protect the gonads. |
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