Objective:To investigate the neuromodulatory effect of pinellia total alkaloids(PTA)on the gamma-aminobutyric acidergic(GABAergic)system in epileptic rats,and preliminarily evaluate the anti-epileptic effect of PTA.Me...Objective:To investigate the neuromodulatory effect of pinellia total alkaloids(PTA)on the gamma-aminobutyric acidergic(GABAergic)system in epileptic rats,and preliminarily evaluate the anti-epileptic effect of PTA.Methods:Ninety-one male Sprague-Dawley rats were randomized to a control group(n=17)or an epileptic group(n=74)using computer-generated random numbers.Status epilepticus(SE)was induced with pilocarpine in the epileptic group.Epileptic rats that survived SE were randomly divided into 4 groups,namely an epilepsy group(n=13),a topiramate(TPM,60 mg/kg)group(n=12),a high-dose PTA(800 mg/kg)group(n=12),and a low-dose PTA(400 mg/kg)group(n=10).Treatments were given intragastrically once daily for 14 days.The control group and epilepsy group received normal saline.Spontaneous recurrent seizures(SRSs)were monitored 8-h daily for 7 days after treatment.Then,the hippocampal formation tissues were collected.GABA level was measured using enzyme-linked immunosorbent assay.Protein and mRNA expression levels of glutamate decarboxylase 65(GAD65),GABA transporter-1(GAT-1),GABA transaminase(GABA-T),and GABAA receptor(GABAAR)α4,α5,γ2 andδsubunits were measured using Western-blotting analysis and quantitative polymerase chain reaction.Results:PTA lowered the incidence and frequency of SRS(both doses vs.the TPM group,P>0.05).Compared with the epilepsy group,PTA increased the levels of GABA(both doses P<0.01)and GAD65(mRNA,800 mg/kg,P<0.01),and suppressed the levels of GAT-1(mRNA,800 mg/kg,P<0.01;400 mg/kg,P<0.05),GABA-T(mRNA,both doses P<0.01),and GABAARδsubunit(protein,800 mg/kg,P<0.05)andγ2 subunit(protein,both doses P<0.01).PTA upregulated the low-expressed mRNA levels of GABAARα5 subunit(400 mg/kg,P<0.01),δsubunit(800 mg/kg,P<0.05),andγ2 subunit(400 mg/kg,P<0.05).Conclusions:PTA regulated the GABAergic system through modulating GABA levels and the expression levels of GAD65,GAT-1,GABA-T,and GABAARα4,α5,γ2 andδsubunits.PTA may exert antiepileptic effects on the pilocarpine-induced epilepsy model.展开更多
文摘Objective:To investigate the neuromodulatory effect of pinellia total alkaloids(PTA)on the gamma-aminobutyric acidergic(GABAergic)system in epileptic rats,and preliminarily evaluate the anti-epileptic effect of PTA.Methods:Ninety-one male Sprague-Dawley rats were randomized to a control group(n=17)or an epileptic group(n=74)using computer-generated random numbers.Status epilepticus(SE)was induced with pilocarpine in the epileptic group.Epileptic rats that survived SE were randomly divided into 4 groups,namely an epilepsy group(n=13),a topiramate(TPM,60 mg/kg)group(n=12),a high-dose PTA(800 mg/kg)group(n=12),and a low-dose PTA(400 mg/kg)group(n=10).Treatments were given intragastrically once daily for 14 days.The control group and epilepsy group received normal saline.Spontaneous recurrent seizures(SRSs)were monitored 8-h daily for 7 days after treatment.Then,the hippocampal formation tissues were collected.GABA level was measured using enzyme-linked immunosorbent assay.Protein and mRNA expression levels of glutamate decarboxylase 65(GAD65),GABA transporter-1(GAT-1),GABA transaminase(GABA-T),and GABAA receptor(GABAAR)α4,α5,γ2 andδsubunits were measured using Western-blotting analysis and quantitative polymerase chain reaction.Results:PTA lowered the incidence and frequency of SRS(both doses vs.the TPM group,P>0.05).Compared with the epilepsy group,PTA increased the levels of GABA(both doses P<0.01)and GAD65(mRNA,800 mg/kg,P<0.01),and suppressed the levels of GAT-1(mRNA,800 mg/kg,P<0.01;400 mg/kg,P<0.05),GABA-T(mRNA,both doses P<0.01),and GABAARδsubunit(protein,800 mg/kg,P<0.05)andγ2 subunit(protein,both doses P<0.01).PTA upregulated the low-expressed mRNA levels of GABAARα5 subunit(400 mg/kg,P<0.01),δsubunit(800 mg/kg,P<0.05),andγ2 subunit(400 mg/kg,P<0.05).Conclusions:PTA regulated the GABAergic system through modulating GABA levels and the expression levels of GAD65,GAT-1,GABA-T,and GABAARα4,α5,γ2 andδsubunits.PTA may exert antiepileptic effects on the pilocarpine-induced epilepsy model.
