Effects Of Total Flavonoids Of Eucommia Ulmoides Oliv.on Mice And Mouse PMS Model

Objective:To observe the effect of total flavonoids of Eucommia ulmoides Oliv.on the perimenopausal period and the depression model of perimenopausal rats and mice,discuss the effect and molecular mechanism of total flavonoids of Eucommia ulmoides oliv,preliminarily to establish the relationship between traditional Chinese medicine,syndrome,biochemistry and gene expression.Methods:The left ovary was removed,80%was removed on the right side with a reservation of 20%,to crate incomplete ovariectomy rat model,after continuous administration for 30 days,the surface features changes of the rats were observed after the last administration;After 2h,the rats were decapitated,centrifugal and determination of serum estradiol?E₂?,testosterone?T?,gonadotropin-releasing hormone?Gn RH?and luteinizing hormone?LH?,follicle stimulating hormone?FSH?and interleukin-2?IL-2?and bone gla protein?BGP?content;The thymus gland,spleen and uterus were removed and their wet weight was calculated;Immunohistochemical method was used to detect the expression of ER in hypothalamus,pituitary gland and uterus,and the expression of androgen receptor AR in hypothalamus and pituitary gland;Using chromatin immunoprecipitation sequencing?CHIP-Seq?determination of ovarian technology by trimethyl histone 3 lysine 4?H3K4me3?for epigenetic changes after treatment,and to observe the correlation of ovarian PI3K-Akt pathway gene expression.The model of complete castration combined with depression was established by ovariectomy and chronic unpredictable stimulation,continuous administration of30d,behavioral tests were performed for the next four days,including the independent activity test,the dark avoidance test?the day before the mouse into the darkroom adaptation?,forced swimming and tail suspension test?throughout the behavioral test of the attention to the surrounding environment to remain quiet?,to observe the ability of mice to adapt to different environments;Behavioral tests were performed to observe the changes of the mice;After the last administration of 2h,the mice were decapitated,detection of serum endocrine indicators E₂,T content;The thymus,spleen and uterus were measured and the wet weight was calculated;The contents of 5-hydroxytryptamine?5-HT?and Dobaamine?DA?in brain homogenate were determined by brain homogenate;Pathological changes of uterus,thymus and spleen were observed under light microscope;Immunohistochemical method was used to detect the expression of ER in hypothalamus,pituitary gland and uterus,and the expression of AR in hypothalamus and pituitary gland.Results:The incomplete ovariectomy rat model was succeed,the rats in the model group decreased level of activity,the spirit of low-spirited,dim and dark hair color;The uterus,thymus,spleen index,serum E₂,T,BGP,IL-2 levels were significantly decreased,FSH,LH,Gn RH water increased significantly;The expression of ER in hypothalamus,pituitary gland,uterus and the expression of AR in hypothalamus and pituitary were significantly decreased.The total flavonoids of Eucommia ulmoides Oliv could significantly or significantly improve the characterization of the perimenopausal rats,the activity and the amount of food was normal;The levels of E₂,T,BGP and IL-2,uterus,thymus and spleen index were increased,reduce the level of FSH,LH and Gn RH;The positive expression of AR in hypothalamus and pituitary and the positive expression of ER in hypothalamus,pituitary gland and uterus were significantly increased;Compared with the model group,the ovary of total flavonoids of Eucommia ulmoides were treated with H3K4me3 antibody,its correlation PI3K-Akt pathway of Igf1r,Fgf11,Gnb2,Bcl-2,Fgf9,Ppp2r2a,Vegfc,Fgf10,Efna3,Foxo3,Cdk6,Prkaa2,Epha2,Sos1,Atf4,Pdgfd gene expression was activated,increased binding to H3K4me3.Complete castration of perimenopausal depression mouse model was succeed,in the model group,the number of spontaneous activity,incubation period,serum E₂ and T levels,the content of 5-HT and DA in brain homogenate,and the indexes of thymus,spleen and uterus were decreased,forced swimming and prolonged immobility time were increased;The hair of model mice was easy to fall off,the color of the hair was grey,the activity was not active and the diet was decreased.Each dose of total flavonoids of Eucommia ulmoides Oliv.could significantly improve the activity and memory ability of perimenopausal depression model mice;The levels of E₂ and T in serum and the contents of 5-HT and DA in brain homogenate were improved;The thymus,spleen and uterus index of mice were improved,and the pathological changes of thymus,spleen and uterus were significantly improved;Prolonged incubation period,reduced the number of shocks,and reduced the immobility time of forced swimming and tail suspension in different degrees;Significantly improved the surface features of model mice.Conclusion:The results showed that the total flavonoids of Eucommia ulmoides Oliv.can stimulate the target organs and target tissues of HPO axis,to improve the biochemical and pathological indexes and the ability to adapt to the internal and external environment of the mice in different degrees,overall regulation of estrogen levels due to the decline in perimenopausal and depression symptoms;Activation of PI3K-Akt pathway in Igf1r,Fgf11,Gnb2,Bcl-2,Fgf9,Ppp2r2a,Vegfc,Fgf10,Efna3,Foxo3,Cdk6,Prkaa2,Epha2,Sos1,Atf4,Pdgfd gene expression,promote the binding with H3K4me3,initially establish the epigenetic system mediated by histone modification in perimenopausal syndrome;At the same time,verified the traditional Chinese medicine total flavonoids of Eucommia ulmoides Oliv.play a role in the body's internal and external environment?HPO axis intervention and behavior?and PMS functional disorder?characterized by syndrome?and organic disease?biochemical pathology?.