ABSTRACT
<p><b>AIM</b>To investigate the relationship between the hypoxia-inducible factor 1(HIF-1) and apoptosis correlative proteins in the cardiomyocyte during hypoxia.</p><p><b>METHODS</b>Rat cardiomyocytes cultured in vitro were divided into normoxia, hypoxia and hypoxic preconditioning groups. The cardiomyocytes in hypoxic preconditioning group were cultured in 1% O2, 5% CO2, 94% N2 for 5 days, 12 h daily before exposed to 0% O2 hypoxia with the hypoxic group. The protein expression of HIF-1alpha, Bcl-2, P53 and Bax in the cardiomyocytes were analysis with Western blot after 48 h 0% O2 hypoxia.</p><p><b>RESULTS</b>With the increment of HIF-1 expression, the Bcl-2 expression was inhibited, the Bax and P53 expression were increased as well during hypoxia. Hypoxic preconditioning could suppress the HIF-1 expression, meanwhile the Bcl-2 expression was elevated, and the Bax expression was decreased.</p><p><b>CONCLUSION</b>HIF-1 may contribute to the regulation of the cardiomyocyte apoptosis with the Bcl-2 family during hypoxia.</p>
Subject(s)
Animals , Rats , Animals, Newborn , Apoptosis , Cells, Cultured , Hypoxia , Hypoxia-Inducible Factor 1, alpha Subunit , Metabolism , Myocytes, Cardiac , Metabolism , Proto-Oncogene Proteins c-bcl-2 , Metabolism , Tumor Suppressor Protein p53 , Metabolism , bcl-2-Associated X Protein , MetabolismABSTRACT
<p><b>AIM</b>To investigate the effects of hypoxia on the secretions of proinflammatory cytokines TNF-alpha and IL-6 and to inquire into the mechanism.</p><p><b>METHODS</b>Separated mice abdominal macrophages which were identified with non-specific esterase dye method, and created the hypoxic cultured model. The levels of TNF-alpha and IL-6 in the medium were determined by ELISA method. The mRNA expressions of TNF-alpha and IL-6 were measured by RT-PCR method. NF-kappaB activation was assayed by Western blot method. Finaly, we added cortone (5 microg/ml) to the medium, then observed the secretion levels of TNF-alpha and IL-6 during hypoxia.</p><p><b>RESULTS</b>The secretions of TNF-alpha and IL-6 from Mphi exposed to hypoxia for 12 h were increased significantly compared with control (P < 0.01). The expressions of TNF-alpha mRNA and IL-6 mRNA were enhanced obviously contrasted with control (P < 0.01). NF-kappaB activation in Mphi nuclei was raised at 2 h during hypoxia and persisted to 5 h. We added cortone to the medium and found no significant change in secretion of TNF-alpha and IL-6 during hypoxia.</p><p><b>CONCLUSION</b>Hypoxia could activate NF-kappaB and make it shift to nucleus which promoted the transcriptions and expressions of TNF-alpha and IL-6.</p>
Subject(s)
Animals , Male , Mice , Cell Hypoxia , Cells, Cultured , Interleukin-6 , Metabolism , Macrophages , Bodily Secretions , Mice, Inbred Strains , NF-kappa B , Metabolism , Oxygen , Metabolism , Tumor Necrosis Factor-alpha , MetabolismABSTRACT
<p><b>AIM</b>To explore the molecular biological mechanism of hemoglobin with high oxygen affinity in Tibetans by determining the sequence of globin cDNA in Tibetans living at high altitude.</p><p><b>METHODS</b>Total RNA was isolated from human bone marrow samples of three Tibetans who live in Qinghai-Tibet plateau. cDNA fragments coding for alpha and beta genes of human hemoglobin were obtained through RT-PCR and were ligated to plasmid pGEM-T easy vectors, and then the ligation liquid were transformed to Escherichia coli and cloned and sequenced. Nucleotide sequences were compared with GenBank data by BLAST method.</p><p><b>RESULTS</b>sequence of a globin cDNA in Tibetans were the same with the registering globin genes in the GenBank, and Hb Abruzzo (beta143 (H21), His- > Arg) gene mutation, a high oxygen affinity beta globin mutation, was found in one Tibetan' beta goblin coding gene (CAC- > CGC).</p><p><b>CONCLUSION</b>This hemoglobin gene mutation may be associated with high altitude adaptation of Tibetans living at high altitude.</p>