Role Of Gap Junction Formed By CX32 And CX43 In Epilepsy And Its Modulating Mechanism

Epilepsy is a kind of common disease in nervous system and it is a chronic disease induced by multi-etiological factors. It is characterized by brain neuron's repeated and excessive super-synchronization discharging, which induces transitory and episodic dysencephalia. Prevalence rate of epilepsy is about 0.46％ and incidence rate is 37/100,000/ year. Drug treatment is a important method to epilepsy. 65% of epilepsy can be cured by drug treatment, however, still 35% of it is not effective to drug. At present, a lot of experiments indicate that epilepsy is related to GJ. However, it is not so clear. that which kind of connexin in forming GJ play important role in epilepsy activity. AIMPresent study made use of advanced experimental method to observe the change of CX32 and CX43 in epilepsy on molecular level so that which kind of connexin in formting GJ play important role in epilepsy was clarified. The finding can define the target that new antiepileptic will act on. CMP, a kind of synthetic specific blocker of GJ, was used to study antiepileptic, which established the basis of the study and therapy of epileptic. Method Wistar rats of male were divided into several groups. There are 25 rats in normal matched control group and operation matched control group respectively and 200 rats in epileptic group induced by KA. Epileptic group was divided into 8 group on the order of 3 hours,6 hours,12 hourses,24 hourses,72 hourses,7 dayses,15 days and 30days. Above each was divided into three groups CMP group, CAR group and saline group, and each group has 5 rats. Above 150 rats were applied to tracing of EEG and surplus 100 rats were applied to Western blot and immunohistochemical analysis according to time course.Method of making epileptic model Rats was fixed on stereotaxic apparatus and take the rhombus slices from the skull center so that cranium, anterior fontanelle and posterior fontanelle was exposured .The position of basiolateral nuclear in right nucleus amygdalae is on the behind of the anterior fontanelle 3.5mm, on the right side of the centerline 4.5mm and under the dura 7.5mm. According to the coordinate of nucleus amygdalae ,KA was injiected into nucleus amygdalae and 1μl PBS was injected as control. Tracing of Electro encephalogram Rats was fixed on stereotaxic apparatus and their body were fixed with adhesive tape.Take the center and lengthways slice on the skull so that cranium , anterior fontanelle and posterior fontanelle was exposured. On the behind anterior fontanelle 2.5mm and on the left side of the centerline 2.5mm, take 3 mm×3mm skull slice away and remove the hard meninges with small dental drill so that cortex sensorimotor area was exposed. Keep the temperature and degree of wetness of the exposed cortex with 3 mm ×3mm filter paper. It is the position that various drugs were infiltrated on. Using Ag－AgCl non-polarizable electrode as channel electrode, the electrode was fixed on the frontal-parietal part and parietal- occipital part by dental cement,under dura mater 0.25 cm. Indifferent electrode was fixed on the tip of nose. All the electrodes were connected with brain electrographic recording. EEG will be recorded 1～1.5h after anesthesia in order to diminish the effect of anesthetic on brain electrographic.Methods of adding drugsAfter pretreated with some drugs(using 500μΜ CMP in CMP group, 10mM carbenoxolone sodium in CAR group and saline in control group), the basical EEG was recorded for half an hour. After that, 4-AP, a kind of inducing Epilepsy drug, was administrated so that EEG induced by Epilepsy was continuously recorded for an hour. The Tracing of EEG in 3h group is different from it in other groups. In 3h group, after basical EEG was recorded for half an hour, the EEG induced CMP, CAR and saline was investigated for half an hour.Main observing markers included in latent period, the amount of epilepsy waves within 15 minutes after the first epilepsy wave, average amplitude and lasting tempo of seizure.The distribution...