Antiepileptic Effect Of Gentiopicroside And Its Effect On Astrocyte-mediated Inflammatory Response

Part I Effects of gentiopicroside on antiepileptic effects and brain inflammation on pilocarpine induced seizures in miceOBJECTIVE: To study the effects of gentiopicroside on antiepileptic effects and brain inflammation on pilocarpine induced seizures in mice.MATERIALS AND METHODS: The model of epilepsy in mice was established by intraperitoneal injection of pilocarpine(PILO,280mg/kg)and randomly divided into Control group,Gent(800mg/kg)group,Model(PILO)group and Gent(800mg/kg)+Model group.SMUP-U4 biological signal processing system was used to detect epileptiform discharges in cerebral cortex;Immunohistochemical staining was used to observed astrocytic activation in brain tissue;Western blot analysis was used to examine the expressions of TLR-4 ?IKK-?? p-IKK-?? I?B-?? p-I?B-?? NF-?B? p-NF-?B? interleukin-1?(IL-1?),interleukin-6(IL-6),and TNF-? in brain.RESULTS: Compared with the model group,Gent(800 mg/kg)significantly reduced the mean amplitude and frequency of EEG;Gent(800 mg/kg)could inhibit the activation of astrocytes after PILO induced seizure in mice compared with the model group;Western blot showed that Gent(800 mg/kg)significantly inhibited protein expression of TLR-4,p-IKK-?,p-I?B-?,IL-1?,TNF-?,IL-6(p<0.05).Moreover,the protein expression of I?B-? was elevated dramatically(p<0.05),but the protein expression of IKK-? and NF-?B showed no prominent difference(p>0.05).CONCLUSION: Gent possesses obvious antiepileptic effect onPILO-induced seizures in mice,and can inhibit the activation and proliferation of astrocytes and the release of inflammatory cytokines in brain of mice;Its mechanism may be related to Gent regulation of TLR-4/NF-?B signaling pathway in the brain of epileptic mice.Part? Effects of gentiopicroside on lipopolysaccharide-induced inflammatory response in astrocyteOBJECTIVE: To investigate the effect of gentiopicroside on lipopolysaccharide(LPS)-induced inflammation response in astrocyte.MATERIALS AND METHODS: The inflammatory response of astrocytes was induced by LPS(1?g/m L)and randomly divided into normal group,model(LPS)group and Gent + LPS group.The levels of inflammatory factors such as IL-1?,TNF-? and IL-6 in the cells of each group were detected by ELISA kit;The expression of TLR-4,IKK-?,p-IKK-?,I?B-?,p-I?B-?,NF-?B and p-NF-?B in each group were detected by Western blot.RESULTS: Compared with the model group,ELISA results show that Gent can significantly reduce the release of IL-1?,TNF-? and IL-6(p<0.05)on LPS-induced inflammation response in astrocyte.Western blot results showed that Gent had no effect on IKK-?,I?B-? and NF-?B protein expression,but significant inhibition of TLR-4,p-IKK-?,p-I?B-?(p<0.05),p-NF-?B protein expression(p<0.05),when compared with the model group.CONCLUSION: Gentiopicroside regulates the astrocyte inflammatory response by regulating TLR-4/NF-?B signaling pathway in astrocytes,which may be the mechanism of anti-epilepsy effect of gentiopicroside.