Research On The Mechanisms Of Pannexins Protein Family After Experimental Intracerebral Hemorrhage In Rats

Partâ… :Expression of Pannexins protein family after experimental intracerebral hemorrhage in ratsObjective: To investigate expression of the Pannexins protein family(including Panx1, Panx2, Panx3) after experimental intracerebral hemorrhage(ICH) in rats.Methods: 42 adult male SD rats were randomly divided into seven experimental groups: sham group(n=6), six hours after ICH group(n=6), 12 hours after ICH group(n=6), 24 hours after ICH group(n=6), 48 hours after ICH group(n=6), 72 hours after ICH group(n=6), a week after ICH(n=6). The rat ICH model was induced by injection of â…¦collagenase(Sigma Chemical Co) into right caudate putamen by stereotactic, Each group of rats were killed in 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 1 week respectively after ICH.The brain sample was removed and keep in 10% formalin solution and liquid nitrogen.The expressions of Pannexins potein family were measured by western blot and immunofluoresence.Results:The protein levels of Pannexins potein family were detected by Western blot.Panx1 was expressed in ICH group and the sham group.the expression of Panx1 was peaked at 48 h in the rat brains of ICH group as compared to that of sham group, The expressions of Panx1 at 48 hr and 72 hr were significantly higher(p <0.01), The expression of Panx1 in 1w was significantly higher(p <0.05). Panx2 and Panx3 were also expressed in ICH group and the sham group, but no significant difference was found in the above index between the sham group and ICH groups(p> 0.05). The protein levels of Pannexins potein family in the rat brains of ICH group were detected by Immunofluorescence doublestaining. the results showed that: the expressions of Panx1 were increased gradually in the rat brains of ICH group, and peaked at 48 hr. but no significant difference was found in the above index between the sham group and ICH groups.Conclusion:1.Pannxins protein(including Panx1, Panx2, Panx3) in rat brains tissue were expressed. 2.The expressions of Panx1 were significantly higher after experimental intracerebral hemorrhage(ICH) in rats,and the above index was peaked at 48 hr.partâ…¡: Research on carbenoxolone(CBX) about apoptosis intervention after experimental intracerebral hemorrhage in ratsObjective:To investigate the effects and mechanisms by carbenoxolone(CBX) intervention on the secondary brain damage and neurobehavioral dysfunction after experimental intracerebral hemorrhage(ICH).Methods: 80 adult male SD rats were randomly divided into four experimental groups: sham group(n=20),ICH group(n=20), low-dose carbenoxolone(10 mg/kg) treatment 1 hour after intracerebral hemorrhage induction(n=20) and high-dose carbenoxolone(20 mg/kg) treatment 1 hour after intracerebral hemorrhage induction groups(n=20).The rat ICH model was induced by injection of â…¦collagenase(Sigma Chemical Co) into right caudate putamen by stereotactic. Part of rats in each group were killed on 48 hr after ICH. Starting neurological function of SD rats at 47 hr after the induction of ICH.Each group of rats were killed on 48 hr after ICH. The brain sample was removed and keep in 10% formalin solution and liquid nitrogen.The expressions of Caspase-3 and albumin were measured by western blot.Detection of cell apoptosis and necrosis ratio By TUNELand FJB.Results: The behavior and activity scores, apoptosis and necrosis percentage of neurons of ICH rats were all significantly higher than those of the sham rats(p<0.01), whilethe above index of ICH+carbenoxolone group were significantly lower than those of the ICH(P<0.05), It is more significant of the above index betweeen ICH group and high-dose carbenoxolone(20 mg/kg) treatment group(p<0.01).Compared with the sham group, the percentage of water content in rat brain of ICH was significantly higher(P <0.05), while the above index of ICH+carbenoxolone group were significantly lower than those of the ICH(P<0.05), It is more significant of the above index betweeen ICH group and high-dose carbenoxolone(20 mg/kg) treatment group(p<0.01).The protein levels of Active-Caspase3 and Albumin were detected by Western blot.Those proteins were significantly higher in the ICH group Compared with the sham group(p <0.05). while the above index of high-dose carbenoxolone(20 mg/kg) treatment group were significantly lower than those of the ICH group(P<0.05).but no significant difference was found in the above index between the low-dose carbenoxolone(20 mg/kg) treatment group and ICH group.Conclusion: carbenoxolone(CBX) may improve the neurobehavioral dysfunction after ICH and decrease the apoptosis and necrosis of neurons.