Effect Of Anisodamine On CB And GFAP Expression In The Ethanol-induced Brain Damage In Rat

Objective: It has been proved that the alcohol abuse and alcoholism can cause both health and social problem severity. Long-term use of alcohol can lead to serious functional and pathologyic abnormalities in many organs, particularly in the brain such as learning and memory impairment, even having dementia Many literatures also demonstrate it can be used for study of neurodegeneration diseases(ND), such as Alzheimer' Disease(AD) using the model of ethanol-induced brain damage in rat Anisodamine, a kind of Chinese medicine, is classified as the drug of cellular protection. The goal of this project is to determine the mechanism of an underlying therapeutic effects of anisodamine on ethanol-induced brain damage including improving of the change of neuropathology, learning and memory processes.Methods: 1. Establishment of the rat model for ethanol-induced brain damage. Healthy male Sprague-Dwley rats were divided into 4 groups with 18 rats each group randomly: group normal saline (NS), group ethanol (EtOH), group anisodamine (An) and group anisodamine plus ethanol (An+EtOH). Animals were received an intraperitoneal (i.p.) injection at dose of 20% ethanol solution (2g/kg/d) daily for eight days. 2. Measurement of alternation behavior in Morris water maze and body weight. 3. Analyses of the changes of Calbindin-D28k(CB) expression and Glial Fibrillaiy Acidic Protein(GFAP) expression in rats hippocampus using method immunohistochemistry and western blotting method. 4. Therapeutic anisodamine's effect on ethanol-induced brain damaged. Results: 1. Test of Morris water maze. (1) The change of latency to platform. There was significantly more prolonged in latency to platform in group EtOH than that in group NS, group An and group An+EtOH, P<0.05. (2) The change of path length. The swimming distance to platform in group EtOH was longer than that in group NS and group An, P< 0.05. There also was longer in group An+EtOH compared to that in group An, but no significant difference between the two groups, P>0.05.2. Immunohistochemistry analysisof CB and GFAP expression in rats hippocampus. (1) The change of CB expression. There was no significant difference among the four groups in hippocampus CA1, CA3 and DG in the mean gray value of CB-positive neurons, P>0.05. (2) The change of GFAP expressioa (D The change of the number of GFAP-positive astrocytes. The number of GFAP-positive astrocytes in group EtOH was increase significantly compared with that in group NS, group An and group An+EtOH in hippocampus CA1 and CA3, P <0.05. An increase in the number of GFAP-positive astrocytes in group An+EtOH was higher than that in group An in hippocampus CA1 and CA3, but there was no significant difference between the two groups, P>0.05. There also was no significant difference in the number of GFAP-positive astrocytes in hippocampus DG among the four groups, P >0.05. (2) The change of the mean area of GFAP-positive astrocytes. The mean area of GFAP-positive astrocytes in group EtOH was significantly higher than that in group NS, group An and group An+EtOH in hippocampus CA1 and in CA3, P<0.05. While there was no differ between group An+EtOH and group An. There was no significant difference in the mean area of GFAP-positive astrocytes among four groups in DQ P> 0.05.3. Western blotting analysis of CB and GFAP expression in rat brain: There was no significant difference in the mean gray values for CB and GFAP of hippocampus, front lobe and cerebellum among four groups, P>0.05. In hippocampus, the mean gray values for CB was higher in group EtOH than that in the other three groups, but there was no significant difference among the four groups.Conclusions: The results indicated 1. There were the decrease in the body weight in rat and significant prolonged in the learning or memory ability in the ethanol-induced brain damage model;2. Ethanol produced significantly increase of the GFAP expression in rats hippocampus CA1 and CA3, while there was no significantly change of the CB expression among the four groups. 3. Anisodamine inhibited significantly the increase of GFAP expression and latency to platform and may be used as a therapeutic agent for ethonal-induced brain injury.