The Effect Of GTPs On The Expression Of NFκB,iNOS And Content Of NO In Hippocampus Of Epileptic Rats

Objective: Epilepsy is one of the most common serious neurologicalconditions,Seizures are caused by sudden, excessive and recurrent electricaldischarges from brain cells. Seizures increased expression of nf-kappa B,cytokines and other inflammatory related molecules, and inflammatory factorsare thought to be closely related to Seizure frequency and survival of theneurons.Despite the existence of a large number of antiepileptic drugs, there iscurrently no cure for epilepsy, and treatment is limited.More than thirtypercent of patients with epilepsy have inadequate control of their seizures bydrug therapy, Furthermore, antiepileptic drugs are associated with a variety ofside-effects and chronic toxicity.In recent years, a great deal of attention hasbeen devoted to the consumption of polyphenols. Green tea polyphenols(GTPs)is the main bioactive compound in green tea, account for up to30%of the dryweight of green tea leaves.which have high anti-inflammatory,antioxidant, andantimutagenic properties in various biological systems.GTPs have been linkedto reductions in the risk of major chronic diseases, such as Parkinson’s,Alzheimer’s and other neurodegenerative diseases.Studies show that GTPsmay decrease the incidence of Alzheimer’s disease and Parkinson’s disease;however, its effect on epilepsy has not been reportedUntil now,Has not yet been reported whether the GTPs is involved inregulating the inflammatory response induced by epileptic seizures.Chronicepileptic model rats kindled by PTZ were established and GTPs wasadministrated.To investigate the effect of GTPs on the changes ofbehavior.Western blotting was used to detected the expression of NF-κBp65and iNOS in hippocampus tissues.Measuring the content of NO inhippocampus tissues by the nitrale reduetase.Methods: The adult male Wistar rats were randomly divided into three groups of twelve animals each group, normal saline control group(NS group),PTZ-induced epilepsy group(PTZ group)and GTPs intervention group(GTPsgroup)．The administration work was conducted between08:30-09:30AM.Prior to the experiments, GTPs and PTZ were dissolved in physiologicalsaline. PTZ was injected intraperitoneally every alternate day in a dose of35mg/kg, while GTPs was injected intraperitoneally daily.NS group received0.9%saline i.p. everyday (3.5ml/kg), PTZ group received saline pretreatmentalong with PTZ every other day,GTPs pretreatment in doses of50mg/kg,respectively in addition to alternate-day PTZ for13injections. In these groups,GTPs was given30min before PTZ; Animals were observed for60min aftereach PTZ administration.The behavioral changes of the rats were observedand assessed by Racine score after the injection．Seizure stage was evaluatedusing the following scale stage0: no response; Stage1: hyperactivity,vibrissae twitching; Stage2: Stage1with head nodding, head clonus andmyoclonic jerk; Stage3: Stage2with unilateral forelimb clonus; Stage4: Stage3with bilateral forelimb clonus; Stage5: Stage4with generalized tonic-clonicseizure (GTCS) with loss of postural control.24h after the last administration,The animals were subsequently killed by decapitation and their hippocampustissues were isolated, Western blotting was used to detected the expression ofnucleus NF-κBp65and cytoplasmic iNOS in hippocampus tissues.Measuringthe content of NO in hippocampus tissues by the nitrale reduetase.Results:1.Results of behavior: The repeated administration ofsubconvulsive PTZ (35mg/kg) induced severe seizures during the13kindlinginjection. From seventh day, rats began to nod,wash their face and andforelimbs began to uplift, showed grade II-Ⅲ of epilepsy.There was nomortality in the GTP group and NS group.Compared with PTZ group,thedegree of epilepsy and the Racine score of GTP group in the19,25days hadobvious difference (p＜0.05，p＜0.001respectively)，No seizures were foundin the NS group.2. Result of nucleus NFκB p65and cytoplasmic iNOS expression levelin hippocampus tissue with Western blot：NFκB p65protein level: NS: (0.93±0.42)；PTZ:(1.57±0.47)；GTP:(1.04±0.40).The expression of NFκB p65protein in PTZ group was markedly increased compared to NS group(P<0.05). The NFκB p65protein in hippocampus of the rats in GTP groupdecreased obviously compared with that in PTZ group (P<0.05). Comparisonsof NF κ Bp65protein between NS group and GTP group were of nosignificant difference (P>0.05). iNOS protein level: NS:(0.44±0.14)；PTZ:(0.73±0.23);GTP:(0.49±0.13).The expression of iNOS protein in PTZgroup was markedly increased compared to NS group (P<0.05). The iNOSprotein in hippocampus of the rats in GTP group decreased obviouslycompared with that in PTZ group (P<0.05). Comparisons of iNOS proteinbetween NS group and GTP group were of no significant difference (P>0.05).3. Content of NO in hippocampus: NS:(3.36±1.19)μmol/gprot, PTZ:(2.09±1.28)μmol/gprot，GTP:(2.26±0.70)μmol/gprot. Compared to the controlgroup, the content of NO was significantly higher in the PTZ group(p<0.05).Compared with the PTZ group, the content of NO was markedly decreased inthe GTP group (p<0.05). There was no statistic difference between the GTPgroup and control group(p>0.05).4. Conclusions: Seizure was induced in rat using intraperitoneal injectionof pentylenetetrazol(35mg/kg).NF-κBp65，iNOS and NO may have relationwith the occurring and developing of PTZ-kindled seizures;GTPs can weakenthe noxious injure for over expression of NF-κBp65，iNOS and NO oncentral nerve system of rat,restrain inflammatory reaction of nerve system.Itmay be one mechanism that GTPs prevent the damages caused byepilepsy.Treatment of PTZ-induced seizure with GTPs in rats significantlyattenuated the maximal seizure classes and the predominant behavioral seizurepatterns compared with the vehicle GTPs is an assistant medicine to cureepilepsy,it can be very useful in clinic application.