Effect Of Kangxianshendan On The Concent Of GABA, Glu In Brains And The Expression Of C-fos In Hippocampus In PTZ-kindled Epilepsy Rats

Objective:Epilepsy is defined as a syndrome of chronic and repeated temporary brain function aberration that commonly occurs in childen who and their parents have heavy burden of body ,mind and economy because of repeated paroxysm and discrimination from society and side effect from taking western medicine for a long time.Therefore,it is a urgent for medical worker resolve this question that develop a cheap and efficient drug having no side effect.Chinese traditional medicine ,which is dominant to resolve this world difficult problem ,is developing continuously.After many years reseaching continuously ,my tutor regards that splenic asthenia is childen,s physiological character , who if have a addicion for eating sweet and greasy food, phleg- matic dampness will form and block in middle energizer easily to form indigestion due to retention of food,which further change into fire-heat that is easily to change phlegmatic damp- ness by consuming body fluids.Considering pathogency of epilepsy due to phlegmatic dampness,indigestion of food and fire-heat interacting each other as both cause and effect,my tutor instituted method of promoting digestion, dissipating phlegm, removmg heat from blood and stopping endogenous wind epilepsy. Under the guidance of that way and combining the studies of the Chinese medicine pharmac- ology ,rationally, Kangxianshendan was made up of powder of Cornu Saigae Tataricae, Ramulus Uncariae cum Unics, Exoca- rpium Citri Rubrum, Retinervus Luffae Fructus, Scorpio, Phcretima,Fructus Cratacgi, Massa Medicata Fermen- tata, Fructus Hordei Gcrminatus, Semen Raphani, Scolopendra, Poria, Radix Rchm- anniae,Radix Scutellariae, Fructus Gardiniae. It has been used in the clinic for ten years and had the dependable curative effect. With combine of Chinese traditional theory and modern technics, this experiment using a model of PTZ-induced epileptic rats observe effect of Kangxianshendan on quantity of GABA, Glu and c-fos in hippocampus so that we probe into the efficacy and the mechanism of Kangxianshendan treating epilepsy in order to find a better way to cure this kind of disease.Method: After adaptability feeding for three day, randomly,elect 10 rats as normal group from 50eight-week-old healthy SD rats (weight: 200±50g). Other rats were caused to have epilepsy by PTZ (32mg/kg) injected intraperitionedlly daily continuously for 28 days. After finishing off injections of PTZ for a week, used a same dose to test, rats that epileptic seizure isⅡor higer grade for five time continuously will be defined as having reached kindling criterion and selected as the objective of study. At last, 36 rats were induced epilepsy successfully by injected intraperitioneally PTZ, 4 rats died, the achievement ratio of making the model is about 90%. Immediately ,36 PTZ-induced epileptic rat were separated randomly into 4 groups as following: Normal control group , Epileptic pattern group , Large-dose Kangxianshendan group , Small-dose Kangxianshendan group , Sodium Valproate group . There are 9 rats in every group, and Large-dose ,small-dose Kangxianshendan group and Sodium Valproate (VPA)group were given intragastic administration respectively with Large-dosage(15g/kg), small-dosage(7.5g/kg) Kangxianshendan and Sodium Valproate (400mg/kg), both normal group and model group were give intragastic administration respectively with saline water (1ml/time).It were given twice a day in all group continuously for 28 days. During period when was making model, observe carefully seizure from rats.Last time, after giving intragstic administration for one hour, kill all rats to mensurate GABA, Glu and c-fos in brains.Results:(1) condition of seizure during period when was making model of PTZ-kindled epileptic rats: On the 11th day, 40 rats used to make PTZ-kindled model occurred seizure different in grade, the 14th day, after having injected PTZ into rats, the convulsion latent period was about 12 minutes and persistence about on time was 8 minutes. More the time of injection, was shorter the latent period was and longer persistence time was. on the 21th and 28th day laten period was respectively 18 and 36 miuntes.The result of making mode was that 36 rats whose epileptic seizure was higher thanⅢgrade 23th day were kindled by PTZ the rest of 4 rats died due to acute seizure.(2)Seizure rate of PTZ-kindled rats effected by medication : Kangxianshendan large-dose group and VAP group exist significant difference (P﹤0.001) compared with model group on rate of resisting seizure. Rate of resisting seizure small-dose was higher than in model group (P﹤0.05) but lower than in high-dose group (P﹤0.05) and VAP group (P﹤0.05),which show Kangxianshendan and SodiumValproate can resist seizure and effect of the former was positive correlation concerned with dosage.(3)Seizure level of PTZ-kinled rats effected by medication:After given medication,on the 7th day,Seizure level in VAP group was lower than in model group (P﹤0.05), which whow Sodium Valproate do the deed quicker than other group. After given medication,on 14th( P <0.05), 21th(P<0.01)and 28th day( P<0.001), large-dose Kangxiansh- endan group and VAP group exist significant difference comp- ared with model group.Seizure level in large-dose group on the 28th day is sighificantly different compared with itself on the 7th and the 14th day (P﹤0.01),small-dose Kangxianshendan group on 28th day is significantly different compared with itself on the 14th and 21th day (P﹤0.05),which show both Kangx- ianshendan and Sodium Valproate can down seizure level. Kangxianshendan smll-dose group and large-dose group on the 14th day was significantly different(P﹤0.05). (4) Latent peri- ond of PTZ-kindled rats effected by medication: Latent perion in model group shorten gradually along with passage of time. Before given medication ,model group was significantly different compared with the 14th ,21th and 28th day (P﹤0.01), the 7th day was significantly different compared with the 28th day(P﹤0.01).Which show kindling effect can be strengthen resulted in that seizure threshold become lower and latent period shorten in a period of time.However,Latent period in Kangxianshendan large-dose and small doss group and western medicine group prolong gradually along with passage of time.Before giren medication and then 7th day ,respectively, Kangxianshendan large-dose group, was significantly different compared with itself on the 21th and 28th day (P﹤0.01). Kangxianshendan small-dose group before give medication exist significantly difference (P﹤0.01)compared with itself on the 21th and 28th day,VPA group before given medication compared with itself on the 7th day exist significenly difference(P﹤0.01),all that show Kangxianshendan and Sodium Valproate can prolong latent period by increasing seizure threshold .(5):Seizure duration of PTZ-kindced rats affected by medication :seizure duration in model group was prolong along with passage of time,in which group seizure duration on the 7th(P﹤0.05)to 28th(P﹤0.01andP<0.001) day was significantly different compared with kangxianshendan large-dose,small-dose group and VPA group .It show that kangxianshendan and Sodium Valproate can shouten seizure duration of PTZ-kindled rats .On the 14th day , Kngxianshendan large-dose group was significantly different from small-dsoe group (P﹤0.05). Compared with itself , before given large-dose kangxiansh- endan was significantly different (P﹤0.05) from on the 14th and the 21th day .All the same (P﹤0.05)was that compared with itself the 7th and the 21th day ,There were significant difference between Kangxianshendan small-dose group itself before given and 14th day(P<0.05).(6)The content of the GABA in the hippocampi of PTZ-kindled rats given medicati- on :GABA in the hippcampus of rats in model group decreased significantly different (P﹤0.05)from that of normal gro- up .After given medication ,GBBA of both Kangxianshendan large-dose and VPA group were increased respectively compared with that of model group and Kangxianshendan small-dose group (P﹤0.05) , not there were significantly difference (P﹤0.05) for normal group comparing with Kangxianshendan large-dose group, but was indistinctive compared with VPA group .(7)The content of the Glu in the hippocampus of PTZ-kindled rats given medication : Glu in the hippcampus of rats in model group increased significantly different (P﹤0.05)from that of normal group .After given medication,GBBA of both Kangxianshendan large-dose and VPA group were decreased respectively compared with that of model group and Kangxianshendan small-dose group (P﹤0.05) , but were there not significantly different (P﹤0.05) .Normal group compared with Kangxianshendan large-dose group was indistinctive compared with VPA group(P>0.05) .(8)The expression of c-fos in hippocampus in PTZ-kindled rats affected by medication : The measurement result of flourescence intensity of c-fos in hippocampus show that model group was significantly different respectively compared with normal- group(P<0.001), large-dose group(P <0.001),small-dose group (P<0.05) and VPA group(P<0.05) , normal group and large-dose group compared respectively with that of small-dose group(P<0.05) and VPA group(P<0.05). There not significantly different (P>0.05) between normal group and large-dose group,the same as small-dose group VPA group (P>0.05) .The flourescenceindex was in full accord with fluorescence in intensity.All of the above show that Kangxianshendan can resist the expression of c-fos in hippocampus in PTZ-kindled rats.Conclusion : Kangxianshendan can resist obviously seizure from PTZ-kindlod rats ,its function mechanism may be that react by reducing the content of Glu and expression of c-fos in the hippocampi and increasing the content of GABA of PTZ-kindled rats.(2)This experiment result provided the experiment of science basis for appling Kangxian- shendan to treat epilepsy in clinic.