| Objective:To detect the expressions of Poly (ADP-ribose)-polymerase1(PARP-1)/apoptosis-inducing factor(AIF) in hippocampus after experimental subarachnoid hemorrhage(SAH) with the use of Z-VAD-FMK in rats. To observe the relationships between the expressions of PARP-1, AIF and Caspase-3and the apoptosis of hippocampus and neurological function by the action of Z-VAD-FMK. To discuss the influence of PARP-1/AIF pathway for apoptosis after SAH with Caspase-inhibiter Z-VAD-FMK.Methods:Male Sprague-Dawley rats were divided into groups:Normal, Sham operation, vehicle (SAH+DMSO)(24h,48h,72h) and Z-VAD-FMK (SAH+Z-VAD-FMK)(24h,48h,72h).SAH model was induced by single blood injection into prechiasmatic cistern. The Z-VAD-FMK or DMSO vehicle was slowly injected into right parococele after the blood infection1h. The neurological functions of each group were assessed with beam balance test at different time point. Then the rats were affused with paraformaldehyde after anesthetizing. And then remove the brain tissue after1h. The inflammation intensity of the hippocampal tissue was observed by hematoxylin and eosin (HE) stains. The expressions of Caspase, PARP-1and AIF measured by Immunohistochemical Method and the apoptotic cells in hippocampus were measured by TUNLE. The neurological functions, Caspase, PARP-1and AIF expressions were compared in each group.Results:1To compared with the normal and sham rats, the neurological function were impacted significantly at6h after SAH and prolonged to48h(p0.05), then there were on difference at72h(p>0.05).Neurological deficits of the Z-VAD-FMK rats showed no difference with the vehicle rats(p>0.05).2The cytoexudation intensity of hippocampus was remarkable severely at24h after SAH, then showed decreasing at48h and72h. Compared with the vehicle rats, the intensity markedly declined of at every time point (p<0.0.5).3The protein expressions of Caspase-3in hippocampus were obviously increased after SAH(p<0.05),and reached a peak level at24h, then deceased regularly but were still higher than that of normal and sham rats at72h(p<0.05). Compared with the Z-VAD-FMK rats, the expression were markedly increasing at24h in the vehicle rats (p<0.05), but no differences in the group at each time point (p>0.05).4The protein expressions of AIF in hippocampus were obviously increased after SAH(p<0.05), and reached a peak level at24h, then deceased regularly but were still higher than that of normal and sham rats at72h(p<0.05). Compared with the vehicle rats, the expression of AIF were markedly higher at24h in the Z-VAD-FMK rats (p<0.05). No differences showed at each time point in the group (p>0.05).5The protein expressions of PARP-1in hippocampus were obviously increased after SAH at24h(p<0.05), and that in the Z-VAD-FMK rats reached a peak level at72h。 The expressions in the vehicle rats reached a peak level at48h, and deceased but were still higher than that of normal and sham rats at72h(p<0.05). Compared with the vehicle rats, the expression of AIF were markedly higher at24h in the Z-VAD-FMK rats (p<0.05). No differences showed at each time point in the group (p>0.05).6There were few apoptosis in hippocampus area in the Normal and Sham rats. After SAH, the apoptosis cell increased significantly at24h(p<0.0.5) both in the Z-VAD-FMK rats and the vehicle rats, then decreased slowly but still keep a higher level(p<0.05). No differences showed at each time point in the group (p>0.05).Conclusion:1The neuro logical deficit of rats showed up immediately after SAH, and then gradually restore. Its function fully recovered at72h after SAH.2The Caspase-pathway and PARP-1/AIF participated in the process of apoptosis after SAH. The Caspase inhibiter Z-VAD-FMK cannot completely stop the apoptosis of hippocampus cell.3By the use of Z-VAD-FMK, it increased the action of PARP-1and AIF than that without Caspase inhibiter, which paralleled to the apoptosis in time. It indicated that PARP-1and AIF might induce the apoptosis with the deficient of Caspase. |
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