1.Involvement of inhibition of nucleus GAPDH over-expression in erythropoietin's reduction of neuronal apoptosis induced by brain ischemia/reperfusion in rats.
Yan-Zhong GUAN ; Ran GUO ; Hong NIAN ; Xiu-Dong JIN
Acta Physiologica Sinica 2012;64(3):269-274
To study whether recombinant human erythropoietin (rhEPO) reduces neuronal apoptosis through inhibiting over-expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in nucleus induced by brain ischemia/reperfusion in rats, 48 adult Sprague-Dawley rats were randomly divided into 3 groups: sham, saline and EPO groups. Animal models of brain ischemia/reperfusion were established by middle cerebral artery occlusion in rats. The effects of EPO on the sizes of ischemia tissue were observed by TTC staining. The over-expression of GAPDH in nucleus was detected by Hoechst-33258 and anti-GAPDH antibody double staining. The neuronal apoptosis in penumbral was detected by Nissl's staining and Hoechst-33258 immunofluorescence, respectively. The results showed that rhEPO treatment (3 000 U/kg, three times daily, i.p.) apparently reduced the sizes of infarct brain tissue in ischemia/reperfusion rats. rhEPO inhibited over-expression of GAPDH in nucleus of apoptotic neurons. In the meantime rhEPO decreased the number of apoptotic neurons in ischemia/reperfusion rats. These results suggest that rhEPO may induced reduction of neuronal apoptosis in penumbra may be through inhibiting over-expression of GAPDH in nucleus of apoptotic neurons induced by ischemia/reperfusion. Reduction of GAPDH over-expression in nucleus may play a pivotal role in EPO inhibiting neuronal apoptosis in cerebral ischemia/reperfusion rats, providing experimental evidence for EPO neuro-protecting effects against ischemia/reperfusion.
Animals
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Apoptosis
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Brain
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enzymology
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pathology
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Brain Ischemia
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pathology
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Erythropoietin
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pharmacology
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Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating)
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metabolism
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Humans
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Rats
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Rats, Sprague-Dawley
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Recombinant Proteins
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pharmacology
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Reperfusion Injury
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pathology
2.Enhanced tyrosine hydroxylase expression in PC12 cells co-cultured with feline mesenchymal stem cells.
Guang Zhen JIN ; Xi Jun YIN ; Xian Feng YU ; Su Jin CHO ; Hyo Sang LEE ; Hyo Jong LEE ; Il Keun KONG
Journal of Veterinary Science 2007;8(4):377-382
Mesenchymal stem cells (MSCs) secrete a variety of neuroregulatory molecules, such as nerve growth factor, brain-derived neurotrophic factor, and glial cell-derived neurotrophic factor, which upregulate tyrosine hydroxylase (TH) gene expression in PC12 cells. Enhancing TH gene expression is a critical step for treatment of Parkinson's disease (PD). The objective of this study was to assess the effects of co-culturing PC12 cells with MSCs from feline bone marrow on TH protein expression. We divided the study into three groups: an MSC group, a PC12 cell group, and the combined MSC + PC12 cell group (the co-culture group). All cells were cultured in DMEM-HG medium supplemented with 10% fetal bovine serum for three days. Thereafter, the cells were examined using western blot analysis and immunocytochemistry. In western blots, the co-culture group demonstrated a stronger signal at 60 kDa than the PC12 cell group (p < 0.001). TH was not expressed in the MSC group, either in western blot or immunocytochemistry. Thus, the MSCs of feline bone marrow can up-regulate TH expression in PC12 cells. This implies a new role for MSCs in the neurodegenerative disease process.
Animals
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Antigens, Surface/metabolism
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Blotting, Western
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Cats/*physiology
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Cell Culture Techniques
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Cells, Cultured
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*Gene Expression Regulation, Enzymologic
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Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating)/metabolism
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Immunohistochemistry
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Mesenchymal Stem Cells/*cytology/metabolism
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Microscopy, Phase-Contrast
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PC12 Cells/cytology/*enzymology
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Rats
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Tyrosine 3-Monooxygenase/*metabolism