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| Protection Effects of Prokaryotic Expressed Radish Phospholipid Hydroperoxide Glutathione Peroxidase and Glutathione on Hydroperoxidemediated Injury in Mouse NIH3T3 Fibroblasts |
| LIU Guan-lan1,LI Tian1,LIU Jin-yuan1,YAN Ze-min2,DUAN Ming-xing1 |
1.School of Life Sciences, Tsinghua University, Beijing100084, China
2.Jiangsu Longliqi Bioscience Co. Ltd., Changshu 215555, China |
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Abstract The radish phospholipid hydroperoxide glutathione peroxidase (RsPHGPx) fusion proteins expressed in Escherichia coli with high efficiency were purified and the antioxidant effects of RsPHGPx together with glutathione on hydroperoxide-mediated injury in mouse NIH3T3 fibroblasts were investigated. The cell viability measurement confirmed that 10μg/ml RsPHGPx, when added to the fibroblasts separately, can only slightly increase the cell viability. However, when combained with glutathione, the antioxidant effects increase dramatically. MDA assay suggests that RsPHGPx can scaveng the lipid peroxides, the results are accordance with the MTT results. ROS flourescence investigations and DAPI experiments show that RsPHGPx can increase the protection effect via inhibiting ROS genreation and this protection help maintain the normal nuclear and membrane conditions of NIH3T3 fibroblasts. This combain experiment of RsPHGPx and glutathione provides experimental evidences for the future production of RsPHGPx.
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Received: 13 April 2010
Published: 25 August 2010
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