ABSTRACT
<p><b>OBJECTIVE</b>To evaluate the translocation of 5-lipoxygenase (5-LOX)) after injuries by transfection with green fluorescence protein (GFP)/5-LOX in PC12 cells.</p><p><b>METHODS</b>PC12 cells were stably transfected with pEGFP-C2/5-LOX (GFP/5-LOX) or pEGFP-C2 vectors (control). After treatment with oxygen-glucose deprivation (OGD), H(2)O(2) or NMDA, GFP/5-LOX localization in the cells was observed under a fluorescence microscope. Wild-type 5-LOX was determined by immunostaining after the treatment.</p><p><b>RESULT</b>In the GFP/5-LOX-transfected cells, GFP/5-LOX was primarily localized in the nucleus; while in the GFP-transfected cells, GFP was localized in both the cytoplasm and nucleus. After OGD and H(2)O(2) treatments, GFP/5-LOX was translocated to the nuclear membrane in 50.6 % and 57.7% cells respectively. However, after NMDA treatment or in GFP-transfected cells, no translocation was observed. Wild-type 5-LOX was distributed in the nuclei and cytoplasm, and all the 3 treatments induced 5-LOX translocation to the nuclear membrane.</p><p><b>CONCLUSION</b>In the PC12 cells stably transfected with GFP/5-LOX, GFP/5-LOX is primarily distributed in the nuclei; the OGD-, H(2)O(2)- and NMDA-induced 5-LOX translocation exhibits different properties.</p>
Subject(s)
Animals , Rats , Arachidonate 5-Lipoxygenase , Genetics , Metabolism , Cell Nucleus , Metabolism , Glucose , Pharmacology , Green Fluorescent Proteins , Genetics , Metabolism , Hydrogen Peroxide , Pharmacology , Microscopy, Fluorescence , N-Methylaspartate , Pharmacology , Nuclear Envelope , Metabolism , PC12 Cells , Protein Transport , Recombinant Fusion Proteins , Genetics , Metabolism , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To determine the effects of dihydroartemisinin (DHA) on the proliferation and apoptosis of rat glioma C6 cells.</p><p><b>METHODS</b>DHA (1~ 125 micromol/L) was added into the cultured C6 cells and incubated for 24, 48 and 72 h. The cell proliferation and viability were determined by trypan blue exclusion assay and 3-(4, 5-dimethylthiazol-2yl)-2, 5 diphenyl tetrazolium bromide (MTT) reduction assay. The apoptosis was detected by Hoechst 33342 staining. The intracellular reactive oxygen species (ROS) was measured by H(2)DCFDA oxidative reaction.</p><p><b>RESULTS</b>DHA 5 ~ 125 micromol/L inhibited the proliferation of C6 cells in concentration- and time-dependent manners, the IC50 at 48 h was 23.4 micromol/L. DHA 5 ~ 25 micromol/L induced C6 cell apoptosis (P<0.05), and 5 ~ 125 micromol/L increased the intracellular ROS (P<0.01).</p><p><b>CONCLUSION</b>DHA inhibits the proliferation and induces the apoptosis of C6 cells; its cytotoxic effect may result from the increase of intercellular ROS.</p>