| ObjectiveIn this study, we want to observe the expression of NKCC1 and GABAARal mRNA and protein in rat hippocampus 3 days after status epilepsy(SE) and the effect of using of sheep BDNF neutralization antiserum (anti-BDNF). We want to investigate the role of NKCC1 in the formation of SE, and the relationship between NKCC1 and GABAARal, and demonstrate the effect of BDNF on modulating the activiation and expression of NKCC1 and GABAARal in SE, and discuss the function of them in epileptic pathogenesis in order to search new target of anti-epileptic treatment.Method130 SD male rats are randomly divided into N.S control group(15 rats), SE group(60 rats) and experiment group(60 rats). Rats of SE group are induced SE by LiC1-PILO i.p. Rats of experiment group received the same treatment as SE group, except for being injected with anti-BDNF 4h before PILO i.p. Rats of both SE group and experiment group are randomly divided into 6 subgroups respectively(15 rats each), and are sacrificed 2h,6h,24h,3d after SE ended. The changes of ethology of two groups during and after SE induced are observed and evaluated. The expression and distribution of NKCC1 and GABAARal in hippocampus are detected with RT-PCR, immunohistochemisty and Westem Blot.Results1. Results of animal model:Compared with the SE group, the achievement ratio of experient group(respectively 90.7% and 75.27%), and mortality rate are lower than SE group (respectively 23.08% and 14.29%). Compared with SE group, the differences of mean dosage of PILO of experiment group for inducing SE(respectively 22.78±6.69mg/ kg and 42.78±6.69mg/kg), mean time needed to induce SE(respectively 41.17±18.94min and 89.72±11.28min), and mean dosage of 10% Chloral Hydrate for ending SE(respectively 0.91±0.05ml/100g and 0.76±0.06ml/ 100g) have significant statistic differences(p<0.01).2. Expression of NKCC1 mRNA and protein are acute substantially up-regulated in SE group within 3d after SE ended, they increase at 2h after SE ended, and come to peak at 6h,then they gradually and slowly recover at 24h, they remain higher than normal at 3d. There are significant statistic differences in all four time points when compared with control (p<0.01). Up-regulation of NKCC1 significantly appears in CA1 and CA3 area after SE. Those of experiment group is with similar distribution but they almost near the normal at 3d after SE ended. Compared with SE group, expression of NKCC1 of the experimental group are significant statistic different in all detected time points(p<0.01).3. The GABAARal mRNA and protein expression of SE group is down-regulated within 24h, but greatly up-regulated at 3d after SE, with significant statistic differences compared with control(p<0.01). The space distribute pattern of GABAARal is great distinct between different areas. It is acute substantially decreased in CA1 and CA3 area since 2h. Compared with control, the GABAARal expression of experiment group is down-regulated at 2h, but up-regulated from 6h to 3d(p<0.01). The distribution pattern of GABAARal of experiment group is similar with SE group. There are significant statistic difference when GABAARal expression of experiment group is compared with SE group at 6h-3d after SE(p<0.01).Conclusion1. The up-regulation of NKCC1 may play an important role in GABAAR-mediated excitation after SE.2. The treatment of anti-BDNF can decrease the action of BDNF protein, delay the onset of SE and reduce the degree and duration of severe SE. |
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