The Mechanism Of Action Of Total Flavonoids From Fenugreek On Hyperplasia Of Mammary Glands Model Of Rats And Mice Based On Neuroendocrine Immune Network

Objective:Base on the neural endocrine immune network,to observe the effect of total flavonoids from Fenugreek on the mammary gland hyperplasia model of mice induced by estradiol benzoate,estradiol benzoate and progesterone,and the mammary gland hyperplasia model of rats induced by estradiol benzoate and progesterone combined with tail clip.Explore the effect and mechanism of total flavonoids from Fenugreek in the treatment of mammary gland hyperplasia.Methods:Experiment 1:The mouse mammary gland hyperplasia model was made by intraperitoneal injection of estradiol benzoate.Established the mouse mammary gland hyperplasia model by intraperitoneal injection of estradiol benzoate.The mice were injected with estradiol benzoate every two days at a dose of 0.5mg/kg for 15times.Divided the model mice into model group,high,medium and low dose group of total flavonoids from Fenugreek.Each group administrat drugs for 30 days.After the last administration,measured the height and diameter of mouse papilla.Detecte the changes of E₂,P,Gn RH,PRL,FSH and LH levels in serum.Calculate the organ indexes of thymus,spleen,uterus and ovary.Observed the pathomorphological changes of mammary gland,thymus and spleen by light microscope.Experiment 2:The mouse mammary gland hyperplasia model was made by intraperitoneal injection of estradiol benzoate and progesterone.The mice were injected with estradiol benzoate every day for 25 days,and progesterone was injected for 5days.On the 5th day of modeling,divide the model mice into model group,high,medium and low dose group of total flavonoids from Fenugree.Each group give corresponding drugs for 30 days.After the last administration,measure the height and diameter of mouse papilla.Measure the levels of E₂,P,PRL,T ER,PR,NE,EPI,IL-2and TNF-αin serum.Calculate the indexes of thymus,spleen,uterus and ovary.Observe the histopathological changes of breast,thymus and spleen by light microscopy.Detecte the expression rate of ERα,PR,Bcl-2 in breast tissue and Gn RHR in pituitary by immunohistochemistry.Experiment 3:The rat mammary hyperplasia model was established by intraperitoneal injection of estradiol benzoate combined with progesterone and tail clipping.Injected estradiol benzoate into the rats werefor 25 days,progesterone for 5days followes.From the first day of modeling,use swallowtail clamp to clamp tail at5cm from the root of rat tail for 20min for 30 days.Divided the rats into groups on the5th day.Given corresponding drugs to each group every day for 30 days.Observed the swelling degree of the rat’s nipple,hair and emotional state every day.After the last administration,Measured the diameter and height of the second pair of papillae on the left side.Measured the levels of E₂,P,FSH,LH,Gn RH,PRL,T,ER,PR,EPI,NE,IL-2,TNF-α,VEGF and b FGF.Calculate the indexes of thymus,spleen,ovary and uterusd.Observed the pathological changes of breast,thymus and spleen.Detected the expression of ERα,PR,Bcl-2,Fas,Fas L in breast tissue,Gn RH in hypothalamus and Gn RH-R in pituitary by immunohistochemistry.Results:Experiment 1:Compared with the model group,each administration group can reduce the swelling and overflow of mouse nipple,and reduce the height and diameter of nipple.Decrease the index of uterus,spleen,ovary and increase the index of thymus.Decreased or increased the level of E₂,Gn RH,PRL,FSH and significantly the level of LH,P in serum.Improved the pathological changes of breast,thymus and spleen in varying degrees.Experiment 2:Compared with the model group,fenugreek total flavonoids can improve the swelling and swelling of mouse papilla.It can significantly reduce the height and diameter of the nipple,improve the biochemical indexes in serum,the protein expression in breast tissue and the pathological changes in organs.Decreased the levels of E₂,PRL,ER,PR,EPI,NE and TNF-αin serum significantly or significantly.Increased the levels of P,T and IL-2.Reduced the expression rate of Bcl-2,ERα,PR in breast tissue and Gn RHR in pituitary significantly.Experiment 3:Compared with the model group,fenugreek total flavone can make the rats’mood relaxed,hair healthy,improve the condition of redness and swelling of the nipple,and reduce the height and diameter of the nipple.Decreased the levels of E₂,FSH,Gn RH,PRL,ER,PR,EPI,NE,TNF-α,VEGF and b FGF in serum significantly or significantly,and increase the levels of P,LH,T and IL-2.Decreased the expression rate of Bcl-2,Fas L,ERα,PR in breast tissue,and increased the expression rate of Fas.Decrease the positive rates of Gn RH in hypothalamus and Gn RHR in pituitary.Improve the pathological condition of breast,thymus and spleen.Conclusions:1.The models of mammary gland hyperplasia in mice and rats were successfully.2.Fenugreek total flavone can improve the mental state,apparent index,biochemical index and protein expression of mammary gland hyperplasia rats and mice in different degrees.It can regulate the neuroendocrine immune network of rats and mice with hyperplasia of mammary gland to achieve the purpose of treating hyperplasia of mammary gland.