| Obj ectiveIn order to determine the change of estrous cycle in ovariectomized female adult SD rats,the animal model of menopausal syndrome was established.Through the changes of body temperature,body weight,uterus index,uteru pathomorphology,serum sex hormone,and neurotransmitters related to reproductive endocrine in hypothalamic arcuate nucleus,the mechanism of Wujiadanfang,an empirical formula,in the treatment of menopausal syndrome,was discussed,which provides experimental basis for clinical application Method1.The rats were randomly divided into the blank,model,sham operation,the high,medium and low dose group of Wujiadanfang,the control group of TCM and Western medicine.The model was made by castration,and was confirmed to be successful by the smear results of vaginal exfoliated cells2.After the model was established,Wujiadanfang,Chinese and Western control drugs were given to the stomach.The changes of behavior and body temperature were recorded.At the end,the body and uterus weight and uterus index were measured.The microscopic morphology of uterus was observed3.Blood was collected from abdominal aorta of rats in each group,and serum sex hormone level was measured by ELISA.The levels of PLC-β1,IP3,IP3R and CaM were measured by ELISA.The expression of KiSS-1,GPR54,GnRH,IP3R-1,PLC-β1 mRNA was detected by PCR,and the expression of KiSS-1,GPR54,GnRH,CaM protein was detected by Western-blotResult1.The effect of temperature:the average abdominal temperature of each group was slightly higher than that of the same group,but it was not statistically significant.Compared with the blank group,the average ear temperature and abdominal temperature in the model group were significantly higher(P<0.01),especially in the third week after operation.Compared with the model group,the average ear temperature and body temperature of the high dose group,the middle dose group,the traditional Chinese medicine group and the western medicine control group decreased significantly(P<0.01);the average ear temperature and the average abdominal temperature of the low dose group decreased significantly(P<0.05)2.The influence of uterus index and pathological morphology after castration,the body weight of rats increased(P<0.01),and the uterus weight decreased(P<0.01),so the uterus index decreased(P<0.01).But the body weight of rats in the sham operation group decreased(P<0.05).Compared with model group,the weight and index of uterus in the high group increased(P<0.01).Under the microscope,it was found that the endometrium and myometrium in the model group were atrophied,the glands were atrophied,and the vascular lumen was obviously narrowed;the endometrium and myometrium were obviously thickened,the glands were enlarged,and the vascular lumen was filled in each group,and the improvement was most obvious in the high group and the western medicine control group.3.The effect of serum sex hormone level:after castration,the serum E2 of model group was lower than that in normal group and sham operation group(P<0.01),and the serum FSH and LH were increased(P<0.01 and P<0.05,respectively),indicating the successful replication of the model.Compared with the model group,the FSH of each group was significantly reduced,the serum E2 of high and middle dose group was increased,and the LH level of high dose group was decreased.4.The effect of neurotransmitters in the arcuate nucleus of hypothalamus:after castration,the expression of KiSS-1,GPR54,GnRH,PLC-β1,IP3R1 mRNA and KiSS-1,GPR54,GnRH,CaM protein were higher than that in the blank group,and Wujiadanfang could down regulate the expression of these neurotransmitter genes and proteins.After castration,the levels of PLC-β1,IP3,IP3R and CaM in hypothalamus decreasedConclusion1.Wujiadanfang can effectively alleviate the temperature rise of rats after castration.Wuj iadanfang can increase the weight of uterus and increase the index of uterus in ovariectomized rats.The effect of high dose group and middle dose group is obvious2.Wuj iadanfang can improve the microscopic morphology of uterus in ovariectomized rats,with endometrial thickening,glands and capillaries significantly increased,especially in the high dose group.3.The serum E2 level of castrated rats can be increased by Wuj i adanfang,but the serum F SH and LH levels be decreased.Wuj i adanfang can increase the estradiol level or promote the transformation of estradiol in castrated rats,so as to improve the menopausal syndrome4.Wujiadanfang can reduce the expression of GnRH mRNA and protein through the influence of KiSS-1,GPR54 and PLC-β1,IP3,IP3R,CaM,which are the upstream regulatory factors of GnRH in hypothalamic arcuate nucleus of castrated rats,so as to coordinate the function of reproductive axis and improve the level of reproductive endocrine in postmenopausal syndrome. |
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