ABSTRACT
<p><b>OBJECTIVE</b>To explore the function of Caspase-3 and p38 MAPK in MMT-induced apoptosis in PC-3M cells.</p><p><b>METHODS</b>After incubation of PC-3M cells with 1 mmol/L MMT, the activity of Caspase-3 was examined. The influence on cells viability of Z-DEVD-FMK, a Caspase-3-specific peptide inhibitor, was also examined. Western blot was used to examine the change of p38 MAPK. The effect on cells viability and Caspase-3 activity of SB203580, a specific inhibitor of p38 MAPK, were also examined.</p><p><b>RESULTS</b>The activity of Caspase-3 increased significantly in MMT-induced apoptosis in PC-3M cells /9P < 0.01), and Z-DEVD-FMK could protect cells from apoptosis (P < 0.01). In this course, the phosphorylation of p38 MAPK could be observed. SB203580 inhibited Caspase-3 activity (P < 0.05) and prevented PC-3M cells from MMT-induced apoptosis (P < 0.05).</p><p><b>CONCLUSION</b>Caspase-3 and p38 MAPK are involved in MMT-induced PC-3M cells apoptosis.</p>
Subject(s)
Humans , Male , Apoptosis , Physiology , Caspase 3 , Metabolism , Caspase Inhibitors , Cell Line , Imidazoles , Pharmacology , Organometallic Compounds , Toxicity , Phosphorylation , Prostate , Cell Biology , Pyridines , Pharmacology , p38 Mitogen-Activated Protein Kinases , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To identify the effect of NEP on Abeta -induced apoptosis in PC12 cells.</p><p><b>METHODS</b>PC12 cells that stably express NEP is generated and the effect of NEP on the process of apoptosis induced by Abeta is analyzed, including the viability of the cells, the production of LDH, ROS and ATP, the activity of Caspase-3.</p><p><b>RESULTS</b>NEP could improve the viability of cells and the production of ATP, inhibit the release of LDH and ROS. In the same time, the activity of caspase-3 descended (P < 0.05). But iNEP had not significant effect on cells apoptosis (P > 0.05).</p><p><b>CONCLUSION</b>NEP has the protective effect on Abeta-induced apoptosis in PC12 cells.</p>
Subject(s)
Animals , Rats , Amyloid beta-Peptides , Pharmacology , Apoptosis , Caspase 3 , Metabolism , Endopeptidases , Genetics , Metabolism , PC12 Cells , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To explore the mechanism of reactive oxygen species (ROS) in manganese chloride (MnCl(2))-induced apoptosis in PC12 cells.</p><p><b>METHODS</b>The model that MnCl(2) induced apoptosis in PC12 cells was established. The apoptotic effect of MnCl(2) on PC12 cells was analyzed with the MTT, the flow cytometry and the DNA fragmentation. The production of ROS and ATP in MnCl(2)-induced apoptosis of PC12 cells was examined. The influence of MnCl(2) on the expression of bcl-xl, bax and the activity of Caspase 3 was also analyzed.</p><p><b>RESULTS</b>MnCl(2) triggered PC12 cells apoptosis in a dose-and time-dependent manner (P < 0.01). The rate of apoptosis was significantly increased (P < 0.01) when MnCl(2) of 2 mmol/L induced PC12 cells for 36 hours. The production of ROS was increased (P < 0.001) and the quantity of ATP was decreased (P < 0.01) in PC12 cells with the same inducement of MnCl(2). The expression of bcl-xl was inhibited and the bax was activated in this process (P < 0.01). Caspase 3 was also activated (P < 0.01).</p><p><b>CONCLUSION</b>MnCl(2) induces apoptosis of PC12 cells, which is related to the increase of ROS, the inhibition of the mitochondria and the activation of Caspase 3.</p>
Subject(s)
Animals , Rats , Adenosine Triphosphate , Apoptosis , Caspase 3 , Metabolism , Chlorides , Toxicity , DNA Fragmentation , Manganese Compounds , PC12 Cells , Reactive Oxygen Species , Metabolism , bcl-2-Associated X Protein , bcl-X ProteinABSTRACT
<p><b>OBJECTIVE</b>To evaluate the effects of small interference RNA (siRNA) on epidermal growth factor-like domain 7 (egfl7) gene expression in human endothelial cell line HUVEC.</p><p><b>METHODS</b>siRNA targeting egfl7 (siRNA1, siRNA2, siRNA3 and siRNA4) was constructed through online design of Amnion company and transfected into human endothelial cell line HUVEC with lipofectamine. The nontransfected cells and cells treated with control siRNA were taken as controls. At 24, 48 and 72 hours post various interventions, cell viability was determined by MTS method as well as LDH and ATP releasing tests. egfl7 expressions at protein and mRNA levels were detected by Western blot and RT-PCR respectively.</p><p><b>RESULTS</b>Cell survival rate, LDH and ATP release were significantly reduced in siRNA treated cells compared to control cells (P < 0.05). Similarly, egfl7 expression at protein and mRNA levels was also significantly reduced in siRNA treated cells (P < 0.01), especially in siRNA1 treated cells.</p><p><b>CONCLUSION</b>siRNA inhibited egfl7 gene expression and cell survival in HUVEC.</p>