| Objective We investigated the expression of ARF6 (ADP ribosylation factors 6) and its upstream effector , GTP-ase activating protein(GAP) centaurin-a1, at the protein level in the temporal lobe of intractable epilepsy(IE) patient brains to explor its plssible role in the genesis of IE.Methods A total of 40 patients with refactory epilepsy were recruited from Epilepsy Prevention and Cure Center of Chongqing, whose specimens came from The First Affiliated Hospital of Chongqing Medical University, Beijing Tiantan Hospital, Xuanwu Hospital of the Capital University of Medical Sciences, and Xinqiao Hospital of the Third Military Medical University. Patients received resection of the anterior temporal lobe.Temporal lobe tissues were collected from these 40 patients for this study. According to the patients' cause of IE, we divided them into two groups: short cause(<10 years) and long cause(>10 years) groups. The control group consisted of temporal lobe samples obtained from 10 patients who underwent neurosurgical intervention due to brain trauma in the Neurosurgical Department of The First Affiliated Hospital of Chongqing Medical University. All controls were diagnosed as brain trauma, and neuropathologists found no abnormality in the slides of specimens of these control patients. The expression of ARF6 and centaurin-al in the anterior temporal neocortex was detected using immunohistochemistry, immunofluorescence and western blotting.Results Positive expression of ARF6 and centaurin-a1 (buffy particles in the cytoplasm of neurons) was observed in the cortex of the temporal lobe in both IE patients and the control group. The mean optical densities of ARF6 for the short cause( < 10 years) group, long cause( > 10年)group and control group are (30.80±4.04) , (28.50±6.45) and (39.50±2.95) , the mean gradation are (135.40±3.15) , (129.70±2.10) and (118.40±1.36), respectively. The two epilepsy groups are significantly higher of ARF6 compared to the control group (P<0.05) respectively by analysis of variance. However, there is no significance between the short cause group and long cause group (P>0.05) . The mean optical densities of centaurin-a1 for the short cause(<10 years) group, long cause(>10 years)group and control group are (24.10±4.17),(25.15±5.32) and (15.50±3.24), the mean gradation are (117.60±5.85), (112.70±5.81) and (150.30±9.42), respectively. The two epilepsy groups are significantly higher of centaurin-a1 compared to the control group (P< 0.05) respectively by analysis of variance. However, there is no significance between the short cause group and long cause group (P>0.05) . In immunofluorescecce, the expression of ARF6 of epilepsy groups is fainter than the control group, whereas, there are stronger centaurin-a1 staining in the epilepsy groups than the control group. Western blot analysis was performed according to the manufacturer's protocol. Samples chosen randomly from the short cause group and long cause group (n=8) and control tissue group (n=6) were cut into small pieces and homogenized in buffer including protease inhibitors. Three groups' ratio of the mean gradation of ARF6 andβ-actin are (0.32±0.05),(0.29±0.05) and (0.39±0.04), The two epilepsy groups are significantly higher compared to the control group (P<0.05) respectively . However, there is no significance between the short cause group and long cause group (P> 0.05) . Three groups' ratio of the mean gradation of centaurin-al andβ-actin are (0.52±0.95),(0.57±0.93) and (0.22±0.13), The two epilepsy groups are significantly higher compared to the control group (P<0.05) respectively. However, there is no significance between the short cause group and long cause group (P>0.05) .Conclusions We found ARF6 and centaurin-a1 are both expressed in neurons.The expression of ARF6 in the temporal lobe of IE is weaker than the control group, whereas, the expression of centaurin-a1 in IE is stronger than the control group, this suggest that these two proteins may participate the invation of IE. |
PDF Full Text Request