| Objective:By setting up an experimental model of pure cerebral concussion(PCC)and observing the expression of IL-1β in the hippocampus of PCC rats and intervention of AH6809(EP2 receptor antagonist),to investigate the pathological changes of IL-1β in PCC rats and the regulation of AH6809.Methods:Under the waking state,rat models of PCC rats were constructed by a self-made "metallic pendulum-striker concussive device" and were randomized into PCC 3 h,12 h,1 d,2 d,3 d and 7 d(n=5 for each time point)groups after injury,subsequently.One normal group(n=5)was used as control.The groups of intervention with AH6809 were randomly divided into 1 d,2 d,3 d and 7 d(n=5 for each time point)groups.One control group(PCC+NS)was setting at the corresponding time point(n=5 for each time point).To observe the immuno-expression,distribution,protein and mRNA levels of IL-1β in the hippocampus after PCC,and the role of the intervention AH6809,the immune-histochemical(IHC)staining(S-P method),Western Blot and RT-PCR were used respectively with anti-IL-1β polyclonal antibody.Results:(一)The PCC group(1)Immunohistochemical staining showed that the expression of the IL-1β was weak in the hippocampus under the normal conditions.After PCC injury,the expression of IL-1β immunoreactivity and the number of the IL-1β immunopositive cells increased gradually at each time point(3 h,12 h,1 d,2 d,3 d)in the hippocampus including CA1～4 and dentate gyrus when compared with the control group,and were strongest at 3 d and down-regulation in 7 d.Compared with the normal control group,there was statistically significant(P<0.05)at four time points except 3 h and 7 d.(2)Results of WB and RT-PCR revealed that the protein and mRNA expression of IL-1β was weak in the hippocampus including CA1～4 and dentate gyrus in normal conditions.After PCC injury,the expression increased gradually is accompanied by a variation with time,reached its peak at 3 d and down-regulation in 7 d,these results were consistent with the SP staining.Statistically significant(P<0.05)find the all time groups of the WB and the 1 d,2 d,3 d groups of the RT-PCR.(二)The PCC+AH6809 group(1)Results of SP staining revealed that the expression of IL-1β immunoreactivity and the number of the IL-1β immunopositive cells appear decline in AH6809 group of 1 d,2 d,3 d and 7 d when compare with the PCC + NS group(P<0.05).(2)Results of WB revealed that the protein level of IL-1β appear decline in different degrees in AH6809 group of 1 d,2 d,3 d and 7 d when compare with the PCC + NS group(P<0.05).(3)Results of RT-PCR revealed that the mRNA level of IL-1β appear decline in AH6809 group of 1 d,2 d,3 d and 7 d when compare with the PCC + NS group(P<0.05).Statistically significant(P<0.05)only find in the 3 d group of the RT-PCR.Conclusion:(1)Under normal physiological condition,there were a small amount of IL-1βpositive cells in the hippocampus.(2)The expression of the IL-1β appear certain regularity of sequential changes in hippocampus and suggest that IL-1β may be involved in pathological changes after PCC injury.(3)The intervention of AH6809 have effective inhibitory role in the IL-1βexpression. |
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