ABSTRACT
BACKGROUND:The oxygen free radicals and apoptosis play an important role in limb ischemia/reperfusion injury, so we can al eviate limb ischemia/reperfusion injury by inhibiting the production of oxygen free radicals and apoptosis. OBJECTIVE:To discuss the application and effect of edaravone on limb ischemia/reperfusion injury in rats. METHODS:Of the 30 female Sprague-Dawley rats, 20 rats were randomly selected to make models of limb ischemia/reperfusion injury by ligating the root of right lower limb with a self-made bal oon cuff at 40 kPa pressure to block blood flow for 4 hours and reperfusing. After success model establishment, they were randomly assigned to two groups. In the edaravone perfusion group, edaravone 3 mg/kg was injected via the left femoral vein at 5 minutes before reperfusion. In the model group and normal group (the remaining 10 rats), an equal volume of physiological saline was given at the same time point. At 24 hours after reperfusion, the right anterior tibial muscle of each group was removed and these ultrastructural changes were observed by transmission electron microscope. Bcl-2 mRNA and Bax mRNA of rat anterior tibial muscle of each group were semiquantitatively detected with the RT-PCR and the ratio of bcl-2/bax was calculated. RESULTS AND CONCLUSION:(1)Electron microscope results:compared with the model group, the muscle fibers were neater;the M line and the N line were clearer;the swel ing of mitochondria was al eviated;the numbers of mitochondria and mitochondrial crista were also increased in the edaravone perfusion group. (2)RT-PCR results:At 24 hours after reperfusion, the relative expression of bcl-2 mRNA and the ratio of bcl-2 mRNA to bax mRNA in right anterior tibial muscle were lower in the model group compared with the edaravone perfusion group (P<0.05). However, relative expression of bax mRNA was greater in the model group than that in the edaravone perfusion group, which were both higher than the normal group (P<0.05). Results indicated that the free radical scavenger edaravone relieved limb ischemia/reperfusion injury by improving the mitochondrial ultrastructure and promoting expression of bcl-2 mRNA and inhibiting expression of bax mRNA, and could provide a new choice for the treatment of limb ischemia/reperfusion injury.
ABSTRACT
BACKGROUND:Recent studies found that some factors play important role in the process of denervated muscle atrophy, especial y the feak-headbox transcription factor, is the key element to regulate the denervated muscle atrophy. OBJECTIVE:To investigate the effect of RNA interference on inhibiting feak-headbox 3a gene expression in vitro. METHODS:The myoblast cel line L6 were cultured in the 6-wel cel culture plates, then pEGFP-N1 and smal interfering RNA recombinant plasmid with the same ratio was transfected under the Lipofectamine2000 mediation to optimize the transfection efficiency of the detection system;2μg smal interfering RNA recombinant plasmid of feak-headbox 3a gene were transfected with myoblast cel line L6 for 48 and 72 hours. RESULTS AND CONCLUSION:At 48 hours after pEGFP-N1 and siRNA recombinant plasmid transfection, a large number of bright green fluorescent displayed under fluorescence microscope with higher transfection efficiency. Real-time quantitative PCR analysis showed that there were significant differences in the sequences of feak-headbox 3a-Ⅰ, feak-headbox 3a-Ⅱ, feak-headbox 3a-Ⅲ, feak-headbox 3a-Ⅳ on feak-headbox 3a mRNA when compared with the control group at 48 and 72 hours after trasfection (Phours after transfection when compared with that at 48 hours after transfection. Western Blot gray analysis showed that there were significant differences in sequences of feak-headbox 3a-Ⅰ, feak-headbox 3a-Ⅱ, feak-headbox 3a-Ⅲ, feak-headbox 3a-Ⅳ on feak-headbox 3a mRNA when compared with the control group at 48 and 72 hours after trasfection (Psignificantly inhibit the fork-head transcription factor feak-headbox 3a gene expression, and the inhibition effect of feak-headbox 3a gene smal interfering RNA recombinant plasmid transfected with the sequence on the mRNA and protein level of feak-headbox 3a is not clear, which can provide new idea for the gene therapy of RNA mediated denervated skeletal muscle atrophy.