Study On Impact Of Wistar-Rats' Reproductive And Endocrine Function Based On Anti-Epileptic Drugs

Experimental purposesEpilepsy is the Paroxysmal brain dysfunction for the sake of Brain neuron populations' excessive discharge,and it is a Chronic brain dysfunction syndrome featuring in a variety of causes and acute,and it is a common Central nervous system disorders in Clinical Pediatrics.Epidemiological survey shows that the prevalence rate for epilepsy in china is 0.70%,the prevalence rate of active epilepsy is 0.46%, patients with epilepsy in our country will be about nine million if estimated per 0.70%.Antiepileptic treatment is still mainly relying on long-term oral anti-epileptic drugs to control outbreak.Antiepileptic therapy calls for rational drug use,pays attention to single-drug treatment.Single-drug treatment is the primary means of epilepsy drug treatment for a long time.Experiments show that about 60%of newly diagnosed patients with epilepsy achieve full recovery after receiving antiepileptic drug monotherapy,at the same time without the side effects that can not be tolerated. But at the same time we have seen in the remaining 30%to 40%of epilepsy patients still required the application of anti-epileptic drugs.Traditional first-line AEDs and new AEDs Are widely used in clinical,Many randomized,double-blind, placebo-controlled trial and Clinical observation of these AEDs have verified the efficacy and safety.However,long-term anti-epileptic drugs can cause a series of side-effects,in recent years,the relationship between antiepileptic drugs and endocrine,especially and sex hormones arouse more and more attention.It has been reported the application of VPA alone can cause polycystic ovary syndrome, hyperprolactinemia,menstrual abnormalities,etc.By OXC's rapid metabolism into the active metabolite in the body (10,11-dihydro-10-hydroxy-carbamazepine),whichdeactivatesv-oltage-gated sodium channels to play a role.Study shows that,either as monotherapy or adjuvant,OXC is valid for children with intractable epilepsy in partial seizure.It also has been reported that it can cause the secretion disorder of sex hormones.As a new antiepileptic drug, LEV has a strong antiepileptic effect,and is valid on the partial and grand seizure. The structure is different when compared with theexisting anti-epileptic drugs.Its mechanism has not yet been fully understood.Recent study shows that the brain synaptic vesicle protein SV2A is the unique role that plays a role of anti-epileptic. Study directed at children shows that,as the joint application,it is valid on children in a variety of types of epilepsy.And the effect is more obvious torefractory epilepsy and partial seizure.At present,there is nothing about its impact on sex hormone-related reports,overseas study shows that LEV could cause Wistar rat serum estradiol,progesterone,estrogen follicle(FSH) concentration to decline,and Testosterone concentration is significantly increased,and Luteinizing hormone(LH) concentration remains unchanged.Now that single-drug therapy can cause changes in hormonesecretion,we hope to learn more about whether the combined application of anti-epileptic drugs would give changes in sex hormone secretion,and what changes have aroused,whether or not it is different with single-drug treatment.As first-line AEDs,Sodium valproate have used in clinical for long time;As new AEDs,Levetiracetam andOxcarbazepine are widely used in children who have epilepsy.Currently,Most of the studies are directed at the traditional AEDs(such as sodium valproate),single drug therapy on the reproductive endocrineeffects,an-d lack of research of combination ones,Furtnermore,most of the current study is directed at adults,reports about children at puberty are rarely found.This experiment compares VPA,OXC,and LEV monotherapy with LEV combined with the first two separately applying to childhood until adulthood in female Wistar rats to observe the effect of combined and single application ofanti-epilep-tic drugs for female Wistar rats' sex hormone secretion.By observing the different groups of female Wistar rats' sex hormone levels,to lay the foundation of the clinical application of anti-epileptic drugs,to predict Female sex hormone levels in children with epilepsy after joint application of anti-epileptic drugs, to provide the scientific basis for the prevention and reduction of side effects of antiepileptic drugs,to Improve the medication compliance,To improve the quality of life of children with epilepsy.Experimental methodsSet up a female Wistar rat model of long-term antiepileptic drug application,60 Healthy female Wistar rats,(age:21-22d,weight:48-58g),which are bought from Shandong university Laboratory Animal Center.The Experimental animals were randomly divided into 6 groups(n=10):normal comparison group(CON group), VPA group(VPA group),OXC group(OXC group),LEV group(LEV group),LEV group(LEV group),LEV+OXC group(LO group),LEV+VPA group(LV group).In accordance with the group,the doses are as follows:CON group 10ml/((kg.d) Saline, VPA group 200mg/(kg.d) VPA,OXC group 100mg/(kg.d) OXC,LEV group 100mg/(kg.d) LEV,LO group 100mg/(kg.d) LEV+100mg/(kg.d) OXC,LV group 100mg/(kg.d) LEV+ 200mg/(kg.d) VPA.Gavage twice daily for 90 days.Weighed once a week during Delivery,dose Adjusting according to body weight.we determine rats' cycle phase and then collect specimens:begin when rats are 3 months,to do vaginal smear with Rats in each group at a fixed time every day(6: 00,14:00,20:00),after Alkaline methylene blue staining solution,we observe the Histological Changes of vaginal smears from Light microscope,to identify the rats' different stages of estrous cycle.At inter-estrous period,weighing,taking blood, 3000 rev/min at 4℃immediately after the blood,getting the serum after 15 minutes' centrifuge,preserved in-80℃refrigerator,determination after specimens collected to eliminate the measurement error,And isolating the ovary, uterus.Determination of rats' ovarian weight:After isolating the ovary,rinsing with normal saline,wiping the surface using absorbent paper,weighting,then fixing by 4%paraformaldehyde,Preparing paraffin sections,and then proceeding to HE staining.And detecting the number of follicles and corpus luteum under the microscope.Determination of Estradiol and progesterone:Detected using the radioimmunoa -ssay method.Results1) Female Wistar rat model of Long-term antiepileptic drug:Experimental animals generally in good condition,Activity,feeding,etc.in good condition,no death cases.2) Indicators Detecting:In single-drug treatment group,ovarian weight decreased in LEV Group when compared with CON group,There is a significant difference(P＜0.05);ovarian weight decreased in OXC Group,There is a significant difference(P＜0.05);other groups have no significant differences when compared with CON group(P＞0.05),ovarian weight changes in joint treatment group have no significant difference when compared with CON group(P＞0.05).3) The number of follicles and corpus luteum:In single-drug treatment group, the numbers of follicles and corpus luteum in each group have no significant differences when compared with CON group(P＞0.05).in the joint treatment group, the number of corpus luteum increases when compared with CON group,There is a significant difference(P＜0.05),the number of follicles decreases,There is a significant difference(P＜0.01),the number of follicles decreases in LV group,There is a significant difference(P＜0.01),There is no significant difference in the number of corpus luteum(P＞0.05).4) The sex hormone levels:In single-drug treatment group,VPA Leads to a lower level of progesterone when compared with CON group,There is a significant difference(P＜0.01),LEV results in lower estradiol levels,There is a significant difference(P＜0.01),And at the same time it leads to a reduction in the level of progesterone when compared with CON group(P＜0.01).Others have no significant differences(P＞0.05).In the joint treatment group,Progesterone levels increases in LO group when compared with CON group,there was a significant difference (P＜0.01),Progesterone levels decreases in LV group,there was a significant difference(P＜0.01).Others have no significant differences(P＞0.05).Conclusions 1) Long-term both single and combined application of antiepileptic drug may cause Disorders of Wistar rats' sex hormone secretion and Ovarian follicular dysplasia.2) Clinical application of anti-epileptic drugs,it is necessary to take into account its antiepileptic effect,but also taking into account its side effects.Innovative and significance1) There are some reports that the traditional AEDs alone can cause the disorders of sex hormone secretion in the domestic,Innovation of this paper is that it firstly does some work in the domestic on combination application of levetiracetam, oxcarbazepine and antiepileptic drugs;it focuses on the impact on Wistar rats' reproductive endocrine function,which has provided first-hand side-effects information of combined clinical antiepileptic drugs application.2) It shows that both single and combined application of antiepileptic drugs Affect Wistar rats' hormone secretion,which provides a scientific basis for further preventing and reducing the side effects of antiepileptic drugs.Study limitations1) The mechanism of Anti-epileptic drugs affecting ovarian follicular developm ent remains for further study.2) The mechanism of Anti-epileptic drugs affecting sex hormone levels remains for further study.3) For the sake of time limitations,the results have not been further combined with clinical.