The neuroprotective effects of danshen extract, magnesium lithospermate B and ouabain

• Main Authors: Ya-Chin Hou, 侯雅琴
• Other Authors: Jason T. C. Tzen
• Format: Others
• Language: zh-TW
• Published: 2008
• Online Access: http://ndltd.ncl.edu.tw/handle/79649484414100423641

碩士 === 國立中興大學 === 生物科技學研究所 === 96 === The cardiac glycoside ouabain has been used to treat heart disease. Cardiac glycosides were recently demonstrated to provide neuroprotection against ischemic stroke. Danshen,which is a traditional Chinese herbal medicine, has been widely used in the treatment of cardiac diseases. Magnesium lithospermate B (MLB) is the major water-soluble compound isolated from Danshen. It was assumed to inhibit the activity of Na+/K+-ATPase like the cardiac glycoside ouabain. It was also demonstrated to provide neuroprotection against ischemic stroke in a brain slice assay model. However, the mechanism underlying the neuroprotective effects of danshen extract (DE), MLB and ouabain is still unknown. We therefore investigated the mechanism underlying neuroprotective effect of those drugs by measuring the cell viability, cell migration and cell volume change. The mouse microglia BV-2 cell, the mouse neuronblastoma N2a cell and the human neuronblastoma SH-SY5Y cell lines were used in this study. In MTT assay, the cell viability had no significant effect after treatment with DE, MLB and ouabain on BV-2 cells, but the survival rate of N2a cells was decreased as the time and doses increased. The viability ratio of SH-SY5Y cells was also decreased at high doses of ouabain or DE. In cell migration assay, the BV-2 cells treated with 1 uM of ouabain for 12 h had higher migration density compared with those of other untreated cells or cells treated with MLB or DE. SH-SY5Y cells treated with 1 uM ouabain for 24 or 48 h, the border of the scratch wounds became irregular, therefore could not be measured. In cell volume change detection, the effect of 1 uM MLB had the highest ratio volume change of SH-SY5Y cells but 1 uM ouabain treatment only increased ratio of cell volume change within 90 min. These results suggested that MLB and ouabain may selectively provide protection on neurons. In order to understand the neuroprotective effect of MLB and ouabain, we established either Aβ25-35 or Fibril Aβ25-35-infected cell culture model by mimicking processes of Alzheimer’s disease. Then, we applied either MLB or ouabain into the Aβ25-35 or Fibril Aβ25-35-infected cell culture for 90 min. MLB or ouabain resulted in selectively reducing the number of BV-2 or SH-SY5Y cells after treated with Aβ25-35 or Fibril Aβ25-35. Therefore, we suggested that MLB and ouabain might possess neuroprotective effects against Aβ contamined cells. The possible mechanism may be mediated by regulating calcium oscillation triggered by the inhibition of Na+/K+-ATPase activity and promotion of cell proliferation via a Ca2+ dependent mechanism.