BACKGROUND: The death of aging
cells is virtually apoptosis. To a certain extent, it can be interpreted as a series of results of gene activities. Therefore, the inhibition of oncogene's
expression can lengthen the life span of cells and delay aging of brain tissues. OBJECTIVE: To study the effect of abstragulus mongholicus bung (AMB) on
apoptosis of nerve cells and the expression of relevant gene in aging
mice brain. DESIGN: Completely randomized design and controlled experiment.SETTING: Department of Biochemistry, School of Basic Medical Sciences of Jiamusi University. MATERIALS: The experiment was conducted at the Experimental Animal Center and Biochemical Laboratory of Jiamusi University from December 2003 to May 2004. Totally 24 healthy Kunming mice were recruited in this study. There were 8 two-month-old mice (young group) and 16 twelve-month-old mice. All the 16 mice were randomized into abstragulus mongholicus bung
group and old
control group with 8 in each group.METHODS: ① AMB group: Mice in AMB group received gastric gavage of water decoction of AMB 10.8 g/(kg·d). Abstragulus mongholicus bung was provided by the Department of Traditional Chinese Medicine of the First Hospital Affiliated to Jiamusi University, and evaluated by Jiamusi Drug Inspection Bureau. Water decoction was prepared with 2 kg/L raw materials. Mice in old control group and
young group were filled with lukewarm boiled water. ② All the animals were treated as above for 30 consecutive days before put to death. Their brains were taken out immediately and the middle parts of the brains were removed to fix with neutral formaldehyde. The remaining brain tissues were made into mitochondria suspension. Content of manganese superoxide dismutase (Mn-SOD) and malondialdehyde (MDA) was determined with xanthosine oxidase method and TBA chemical colorimetry. Apoptotic cells (cells with yellow nuclei were positive ones) were assayed with in situ end-labeling (ISEL) and expression of bcl-2 gene was assayed with immunohistochemical method. The cells stained brown were positive ones. A total of 400 cells were counted under the 400× microscope. We graded the samples according to the percentage of the positive cells: the number of positive cells < 5% - 5%-10% + 11%-50% > 51% . ③ Grade and quantitative data were compared with rank sum test and t-test. MAIN OUTCOME MEASURES: Effects of AMB on the rate of neuronal apoptosis, the activity of Mn-SOD, the concentration of MDA in mitochondria, and the intensity of the expression of bcl-2 gene.RESULTS: Totally 24 mice entered the final analysis. ① Content of Mn-SOD was higher in young group and AMB group than in old control group (P < 0.05). ② Concentration of MDA and apoptotic rate in young group and AMB group were lower than those in old control group (P < 0.01). ③ Expression of bcl-2 gene was significantly different in young group and AMB group from that in old control group (P < 0.01). CONCLUSION: AMB is found to be able to obviously inhibit neuronal apoptosis in aging mice brain by affecting the activity of Mn-SOD, the concentration of MDA and the expression of bcl-2.
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