Study of miRNA-212 regulating BDNF/TrkB signaling pathway in epileptic neuron model
<p><strong>Objective</strong> To study the effect of epileptic neuron model after transfection of microRNA (miRNA)-212 on brain-derived neurotrophic factor (BDNF)/tyrosine protein kinase B (TrkB) signaling pathway. <strong>Methods</strong> The primary rat hippocampa...
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| Format: | Article |
| Language: | English |
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Tianjin Huanhu Hospital
2014-11-01
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| Series: | Chinese Journal of Contemporary Neurology and Neurosurgery |
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| Online Access: | http://www.cjcnn.org/index.php/cjcnn/article/view/1077 |
| _version_ | 1828517442278653952 |
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| author | Hao CAI Qiu-jing WU Yan-xia ZHU Wen ZHAO Bin LI Yi-jun SONG |
| author_facet | Hao CAI Qiu-jing WU Yan-xia ZHU Wen ZHAO Bin LI Yi-jun SONG |
| author_sort | Hao CAI |
| collection | DOAJ |
| description | <p><strong>Objective</strong> To study the effect of epileptic neuron model after transfection of microRNA (miRNA)-212 on brain-derived neurotrophic factor (BDNF)/tyrosine protein kinase B (TrkB) signaling pathway. <strong>Methods</strong> The primary rat hippocampal neurons were cultivated in vitro for 7 d and were randomly divided into 8 groups: control group, epilepsy group, control + BDNF group, epilepsy + BDNF group, control + miRNA-212 group, epilepsy + miRNA-212 group, control + miRNA-212 + BDNF group, epilepsy + miRNA-212 + BDNF group. Epilepsy model of hippocampal neurons were established by being exposed to Mg<sup>2+</sup> free extracellular fluid for 3 h. And then the neurons were put back into the normal extracellular fluid of magnesium for 2 h and slow virus diluent was dropwise added for transfection, so that miRNA-212 lentiviral vector was structured. Protein was extracted after 48-72 h. BDNF was added into media 10 min before protein was extracted. Immunofluoresence double staining, patch clamp technique and Western blotting were used to observe the effect of miRNA-212 transfection on BDNF/TrkB signaling pathway. <strong>Results</strong> After BDNF was injected, compared with the control group and epilepsy group, the phosphorylated TrkB (pTrkB)/TrkB value was significantly higher in control + BDNF group and epilepsy + BDNF group (<em>P</em> = 0.001), suggesting the BDNF/TrkB signaling pathway was activated. After transfection of miRNA-212, the pTrkB/TrkB value in epilepsy + miRNA-212 + BDNF group was significantly lower than that in epilepsy + BDNF group (<em>P</em> = 0.001), suggesting the BDNF/TrkB signaling pathway was suppressed. <strong>Conclusions</strong> When hippocampal neurons were transfected miRNA-212, the BDNF/TrkB signaling pathway was suppressed, but BDNF could activate the BDNF/TrkB signaling pathway.</p><p> </p><p>doi: 10.3969/j.issn.1672-6731.2014.11.010</p> |
| first_indexed | 2024-12-11T18:41:40Z |
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| id | doaj.art-8202420e777345bf8f0c8f8f3196838a |
| institution | Directory Open Access Journal |
| issn | 1672-6731 |
| language | English |
| last_indexed | 2024-12-11T18:41:40Z |
| publishDate | 2014-11-01 |
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| series | Chinese Journal of Contemporary Neurology and Neurosurgery |
| spelling | doaj.art-8202420e777345bf8f0c8f8f3196838a2022-12-22T00:54:36ZengTianjin Huanhu HospitalChinese Journal of Contemporary Neurology and Neurosurgery1672-67312014-11-0114119729781072Study of miRNA-212 regulating BDNF/TrkB signaling pathway in epileptic neuron modelHao CAI0Qiu-jing WU1Yan-xia ZHU2Wen ZHAO3Bin LI4Yi-jun SONG5Department of Neurology, Tianjin Medical University General Hospital, Tianjin 300052, ChinaGrade 2007, 7-year Graduate School, Tianjin Medical University, Tianjin 300070, ChinaDepartment of Neurology, Tianjin Haibin People's Hospital, Tianjin 300280, ChinaDepartment of Geriatrics, Tianjin Medical University General Hospital, Tianjin 300052, ChinaDepartment of Neurology, Tianjin Haibin People's Hospital, Tianjin 300280, ChinaDepartment of Neurology, Tianjin Medical University General Hospital, Tianjin 300052, China<p><strong>Objective</strong> To study the effect of epileptic neuron model after transfection of microRNA (miRNA)-212 on brain-derived neurotrophic factor (BDNF)/tyrosine protein kinase B (TrkB) signaling pathway. <strong>Methods</strong> The primary rat hippocampal neurons were cultivated in vitro for 7 d and were randomly divided into 8 groups: control group, epilepsy group, control + BDNF group, epilepsy + BDNF group, control + miRNA-212 group, epilepsy + miRNA-212 group, control + miRNA-212 + BDNF group, epilepsy + miRNA-212 + BDNF group. Epilepsy model of hippocampal neurons were established by being exposed to Mg<sup>2+</sup> free extracellular fluid for 3 h. And then the neurons were put back into the normal extracellular fluid of magnesium for 2 h and slow virus diluent was dropwise added for transfection, so that miRNA-212 lentiviral vector was structured. Protein was extracted after 48-72 h. BDNF was added into media 10 min before protein was extracted. Immunofluoresence double staining, patch clamp technique and Western blotting were used to observe the effect of miRNA-212 transfection on BDNF/TrkB signaling pathway. <strong>Results</strong> After BDNF was injected, compared with the control group and epilepsy group, the phosphorylated TrkB (pTrkB)/TrkB value was significantly higher in control + BDNF group and epilepsy + BDNF group (<em>P</em> = 0.001), suggesting the BDNF/TrkB signaling pathway was activated. After transfection of miRNA-212, the pTrkB/TrkB value in epilepsy + miRNA-212 + BDNF group was significantly lower than that in epilepsy + BDNF group (<em>P</em> = 0.001), suggesting the BDNF/TrkB signaling pathway was suppressed. <strong>Conclusions</strong> When hippocampal neurons were transfected miRNA-212, the BDNF/TrkB signaling pathway was suppressed, but BDNF could activate the BDNF/TrkB signaling pathway.</p><p> </p><p>doi: 10.3969/j.issn.1672-6731.2014.11.010</p>http://www.cjcnn.org/index.php/cjcnn/article/view/1077EpilepsyHippocampusBrain-derived neurotrophic factorProtein-tyrosine kinasesMicroRNAsTransfectionImmunoblotting |
| spellingShingle | Hao CAI Qiu-jing WU Yan-xia ZHU Wen ZHAO Bin LI Yi-jun SONG Study of miRNA-212 regulating BDNF/TrkB signaling pathway in epileptic neuron model Chinese Journal of Contemporary Neurology and Neurosurgery Epilepsy Hippocampus Brain-derived neurotrophic factor Protein-tyrosine kinases MicroRNAs Transfection Immunoblotting |
| title | Study of miRNA-212 regulating BDNF/TrkB signaling pathway in epileptic neuron model |
| title_full | Study of miRNA-212 regulating BDNF/TrkB signaling pathway in epileptic neuron model |
| title_fullStr | Study of miRNA-212 regulating BDNF/TrkB signaling pathway in epileptic neuron model |
| title_full_unstemmed | Study of miRNA-212 regulating BDNF/TrkB signaling pathway in epileptic neuron model |
| title_short | Study of miRNA-212 regulating BDNF/TrkB signaling pathway in epileptic neuron model |
| title_sort | study of mirna 212 regulating bdnf trkb signaling pathway in epileptic neuron model |
| topic | Epilepsy Hippocampus Brain-derived neurotrophic factor Protein-tyrosine kinases MicroRNAs Transfection Immunoblotting |
| url | http://www.cjcnn.org/index.php/cjcnn/article/view/1077 |
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