Effects Of Modified Lou Shi Medicinal Broth On Follistatin And Activin In Deficiency Of Kidney-yin Polycystic Ovary Syndrome Model Mice

Objective Modified lou shi medicinal broth is one experiential recipe developed by my teacher,Which could treatment polycystic ovary and kidney yin deficiency.Aim of study is to preliminarily explore the action mechanism of MLSMB through observe the general condition of mice,the changes of ovary morphology and the levels of serum sex hormones,follistatin(FS)and activin(ACT)through animal experiments.In addition,the maximum dosage of MLSMB was given,To provide experimental basis for safe drug use in clinic.Methods(1)54 female ICR mice,Adaptive feed two day,randomly divided into 6 groups,9mice in each group: blank group,deficiency of kidney-yin PCOS model group,MLSMB high dose,medium dose,low dose group and Diane-35 group.Using dehydroepiuandrosterone sulfate joint thyroxin and reserpine method to establish deficiency of kidney-yin polycystic ovary syndrome in mice model.Except blank group and model group,each group was given the corresponing drug for 40 day.Weigh daily regularly,observe and record the amount of eat and drink,dry wet degree of defecate,hair removal,arch back,autonomic activities,etc.And vaginal smear was performed daily to observe the changes in exfoliated cells after vaginal opening in mice from 28 days old.After experiment,we killed the mice and exucleation and removal of blood immediately,the ovarian and uterine tissues were taken and weighed.The ovary were slced and observed by optical microscope.Detecting the levels of serum luteinizinghormone(LH),testosterone(T),follicle-stimulating hormone(FSH),estradiol(E2),FS,ACT,cyclic adenosine monophosphate(c AMP)and cyclic guanosine monophosphate(c GMP)by using enzyme linked immunosorbent assay(ELISA).(2)Ten 20 g female ICR mice were fed adaptively on2 nd and fasted for 12 hours.They were given the highest concentration of MLSMB(concentrated to 0.4ml /10g).The mice were given 4 times in 24 hours,each time at an interval of 8:00、11:00、14:00、17:00,3 hours after administration.The animals were observed for 7 days and their responses were recorded.Results(1)Compared with the blank group,the model group showed instability,wasting,disorder of estrus cycle,weight decreased,the difference has statistically significant(p<0.05),the weight of ovary and the ovarian index was significantly increased(P<0.05),the serum T,LH levels were significantly increased and E2,FSH were significantly decrease(P<0.01),the serum c AMP/c GMP was increased(P<0.01),the serum FS was increased(P<0.01)and ACT was decreased(p<0.05),and the ovarian polycystic changes.(2)The mice in the medication group did not appear unstable,wasting,and gradually recovered the regular estrous cycle,the weight of the mice was higher than that of the model group,the weight of ovary and the ovarian index were lower than those of the model group(p<0.05).(3)Sex hormone in MLSMB high does,medium dose and Diane-35 group compared with model group serum T and LH were lower,serum E2 and FSH were higher(p<0.05).(4)c AMP/c GMP in MLSMB high dose,medium dose,low does and Diane-35 group is lower than that in model group.(5)MLSMB high does,medium dose and Diane-35 group compared with model group,serum of FS was lower,ACT was higher(p<0.01 or p<0.05).(6)Maximum dose of MLSMB did not cause death in mice,and general situation of mice were well,After dissection,there was no abnormality in heart,lung,spleen,liver,kidney and other organs observed with the naked eye.Conclusion(1)The model mice induced by DHEA have the characteristics of estradiol decrease,hyperandrogenemia,hyper LH and ovarian polycystic changes.(2)The model mice induced by DHEA combined with thyroxine and reserpine can successfully make deficiency of kidney-yin,such as unstable,weight loss,increase of c AMP / c GMP and so on.(3)DHEA combined with thyroxine and reserpine can successfully make deficiency of kidney-yin PCOS model in mice.(4)MLSMB could improve the whole body condition of the model mice with polycystic ovary syndrome,Maintain regular cycle of regulation,increase body weight,lower ovary index,and complete follicular morphology.(5)MLSMB could effectively regulate the level of sex hormone,reduce the ratio of serum c AMP/c GMP,improve the general state of PCOS mice with kidney yin deficiency,decrease the content of FS and increase the content of ACT,It is speculated that this link may promote follicle development.(6)MLSMB is no obvious side effects,can be used safely in clinic.