THE EFFECTS OF TOCOTRIENOL RICH FRACTION ON CHRONIC RESTRAINT STRESSINDUCED CHANGES IN THE HIPPOCAMPUS OF MALE RATS

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Date
2012
Authors
SAIFUL BAHRI, TALIP
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Pusat Pengajian Sains Perubatan Universiti Sains Malaysia
Abstract
Tocotrienol is a member of natural vitamin E that consists of 4 chemically distinct forms: α-, β-, γ- and δ-tocotrienol. Tocotreinol has anti oxidant capacity. Several studies showed the potential protective effect of tocotrienol on stress related organ injury. However the role of tocotrienol on hippocampus after exposure to stress is still largely unknown. Stress is one of the psychological problems which affects most of the individuals in their daily life. Stress may damage the brain especially the hippocampus if it occurs for a long period of time. It is a well known fact that the oxygen radical generated by stress and excess of corticosterone secreted during prolong stress are hazardous on human wellbeing. The purpose of this study was to examine the neuroprotective effect of tocotrienol on hippocampus under restraint stress by observing the histopathological and histomorphometric changes and quantifying the serum corticosterone level in rats after exposure to stress for 21 days. Thirty six male Sprague Dawley rats aged 5 weeks old were divided into four groups: control, restraint stress, tocotrienol and restraint stress treated with tocotrienol. Restraint stress and restraint stress treated with tocotrienol groups were exposed to 5 hours of restraint stress daily for 21 days. Tocotrienol group and restraint stress treated with tocotrienol group received 200 mg/kg body weight of tocotrienol rich fraction (TRF) by oral gavage daily. Control group and restraint stress group received normal saline by oral gavage. Body weight was monitored daily. At the end of experimental period, the rats were sacrificed and the sections of hippocampus were sampled by adopting the systemic random sampling method and were stained with hematoxylin and eosin, cresyl fast violet and immunohistostaining using anti Ki-67 antibody and anti GAP-43 antibody. The serum collected was assessed for corticosterone level by ELISA. Data was analysed by using, PASW statistical software (version 18). The study had found that the level of corticosterone was significantly higher in the restraint stress group as compared to control group and tocotrienol group and this was reflected in the weight of the adrenal gland mass. However, TRF supplementation failed to alter the corticosterone level and the adrenal gland mass. The study also showed the number of proliferative cells in the subgranular zone of the dentate gyrus in the restraint stress group as labelled by Ki- 67 antibody was significantly lower as compared to control group. However, TRF supplementation failed to provide protection against the effect of stress on cell proliferation in the dentate gyrus. The histopathological assessment showed no significant difference in granular cell and pyramidal cell counts between all group comparisons. The histomorphometric assessment also showed no significant different of hippocampal thickness and optical density of GAP-43 in all groups. Therefore the present study shows that TRF supplementation is not effective to prevent the changes in the hippocampus after exposure to chronic restraint stress.
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CLINICAL ANATOMY
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