Study On The Protective Effect Of P2Y12 Receptor Antagonist-PSB0739 On Postepileptic Brain Injury In SD Rats

Objective: To investigate the protective effect of P2Y12 receptor antagonist--PSB0739 on postepileptic brain injury in SD rats.Methods: Thirty 21-day old male SD rats were randomly divided into 6 groups: control group,pentylenetetrazole(PTZ)group,PTZ + 0.1mg/kg PSB0739 group,PTZ + 0.3 mg/kg PSB0739 group,PTZ + 1mg/kg PSB0739 group and PTZ + 2 mg/kg PSB0739 group,with 5 rats in each group.The SD rat model of epilepsy was established through intraperitoneal injection of PTZ,and successful modeling was identified as rats with Grade IV-V behaviors according to the Racine scoring system.Rats in the control group were injected with normal saline,and then administered with intraperitoneal injection of PSB0739 of different doses(0.1 mg/kg,0.3 mg/kg,1 mg/kg and 2 mg/kg)2 hours before PTZ-induced epileptic seizure.The rats were sacrificed 24 hours after the epileptic seizure,and hippocampus tissues were collected.Hematoxylin-eosin(HE)staining was carried out for the observation of pathomorphological changes of brain tissues;Western blot was performed for the detection of IL-1β protein expressions in the hippocampus tissues;and qRT-PCR was used for the detection of IL-1β mRNA expressions in the hippocampus tissues.Results:1.According to the Racine scoring system,significantly lower epileptic seizure grade was observed in the PTZ + 1 mg/kg PSB0739 group and the PTZ + 2 mg/kg PSB0739 group when compared with the PTZ group(all P<0.05);2.After the brain hippocampus tissues were performed with HE staining,neuronal necrosis(nuclear pyknosis)and cellular edema were observed in the PTZ group,while the neuronal necrosis(nuclear pyknosis)and the cellular edema were significantly alleviated in the PSB0739 drug intervention groups;3.Western blot: the IL-1β protein expression in the PTZ group increased significantly compared with the control group(P<0.05),while the IL-1β protein expression in the PTZ + 2 mg/kg PSB0739 group decreased significantly compared with the PTZ group(P<0.05);4.qRT-PCR: the mRNA level of IL-1β in the PTZ group was significantly increased when compared with the control group(P<0.05),while the mRNA levels of IL-1β in PTZ + 0.1 mg/kg PSB0739 group,PTZ + 0.3 mg/kg PSB0739 group,PTZ + 1 mg/kg PSB0739 group and PTZ + 2 mg/kg PSB0739 group were significantly decreased when compared with PTZ group(P<0.05);Moreover,the inhibitory effect of PSB0739 on IL-1β mRNA expression was in a dose-dependent manner,with 1 mg/kg PSB0739 and2mg/kg PSB0739 > 0.3 mg/kg PSB0739 > 0.1 mg/kg PSB0739,respectively.However,there was no significant difference in the inhibitory effect between 1 mg/kg PSB0739 and 2 mg/kg PSB0739 on the IL-1β mRNA expression.5.Correlation analysis: the mRNA and protein expression levels of IL-1β in hippocampus tissues of rat model of PTZ-induced epilepsy were positively correlated with the epileptic seizure grade,respectively(r =0.854,P<0.05;r = 0.654,P<0.05).Conclusions:1.The IL-1β expression level is positively correlated with the degree of postepileptic brain injury in rat.2.PSB0739 may have a protective effect on postepileptic brain injury by inhibiting the increase of inflammatory mediator IL-1β and neuronal necrosis.