ABSTRACT
Aim To explore whether edaravone (EDA), a novel free radical scavenger, protects H9c2 cardiac cells against doxorubicin ( DOX )-induced car-diotoxicity. Methods H9c2 cells were treated with 5μmol·L-1 DOX to establish a model of DOX cardio-toxicity. Cell viability was examined by cell counter kit ( CCK-8 ) . Changes in morphology and amount of ap-optotic cells were detected by Hoechst 33258 staining;intracellular level of reactive oxygen species ( ROS ) was measured by DCFH-DA staining and photofluorog-raphy;mitochondrial membrane potential ( MMP) was observed by rhodamine 123 ( RH123 ) staining and photoflurograph; the expression level of caspase-3 was determined by Western blot assay. Results Pretreat-ment of H9 c2 cells with 20 , 40 and 80 μmol · L-1 EDA for 60 min markedly inhibited cytotoxicity in-duced by 5 μmol · L-1 DOX, respectively, as evi-denced by an increase in cell viability. The protective effect induced by 40 μmol · L-1 EDA was maximal. Pretreatment of H9 c2 cells with 40 μmol · L-1 EDA for 30 , 60 , 90 and 120 min significantly attenuated DOX-induced cytotoxicity, respectively, having a max-imal protection at 60 min. Furthermore, pretreatment of H9 c2 cells with 40 μmol · L-1 EDA for 60 min be-fore exposure to 5 μmol · L-1 DOX for 24 h obviously reduced cardiac injuries, as evidenced by decreases in the DOX-induced intracellular ROS generation, num-ber of apoptotic cells, and expression of cleaved caspase-3, as well as loss of MMP. Conclusions EDA can protect H9 c2 cardiac cells against DOX-in-duced cardiotoxicity, this protection may be associated with inhibition of ROS production and preservation of MMP.
ABSTRACT
Objective To investigate the effects of mitochondrial DNA (mtDNA) on the pathopoiesis mechanisms of paraquat poisoning in vitro.Methods Firstly,the survival rate of A549 cells (human type Ⅱ alveolar epithelial cells) was measured with cell counting kit-8 after exposure to paraquat.Afterwards,the concentration of mtDNA in supernant of culture medium for culturing A549 and the chauge of mitochondrial membrance potential were detected with absolute quantitative PCR and confocal laser microscopy,respectively.Then,The chemotactic activity of mtDNA in peripheral blood mononuclear cells (PBMC) and neutrophils (PMN) were detected by transwell chemotaxis,and the subtype of chemotactic cells was measured with flow cytometry.Meanwhile,the role of mtDNA in vascular permeability was measured by using Xcelligence system and in vitro using vascular permeability kits.Finally,the effects of mtDNA in cell proliferation were to verify.Results The 50％ of lethal concentration (LD50) of paraquat for A549 was 600 μmol/L.Cell viability and concentration of mtDNA following challenge of PQ revealed in a concentration-and time-dependent manner (P ＜ 0.05).The mtDNA had a power in aggregating PBMC nonspecifically,but there was no effect on the vascular permeability was found.Moreover,the proliferation of human fibroblasts was not stimulated directly by mtDNA,but TGF-β1 (transforming growth factor-beta 1),a major pro-fibrotic factor,was increased compared to control group (P ＜ 0.05).Conclusions The mtDNA could play an important role in the inflammatory and proliferation responses to paraquat poisoning.
ABSTRACT
Objective To study topiramate′s neuroprotection on primary cultured hippocampal neurons which were injuried by glutamate and its mechanism.Methods The primary cultured hippocampal neurons were made as the research object,and excitotoxicity model was exected with glutamate.Hippocampal neuron survival was assessed by MTT method and hippocampal neuron mitochondrial membrance potential(MMP) was evaluated by fluorescence microscope and flow cytometry.Results Hippocampal neuron survival of normal control was(98.4?0.8)%,and the survival of glutamate model group was(59.6?3.2)%,at the same time,two topiramate′s groups′ cell survivals was(74.1?0.5)% and(79.2?3.4)%,and topiramate with two levels all could obviously increase hippocampal neuron survival((P