The cytotoxic effect of methylglyoxal on BDNF and TrkB expression in rat hippocampal neurons
Hong Zhou, Zhijun Zhang
Abstract
Hong Zhou, Zhijun Zhang
Abstract
Objective To investigate the mechanisms of methylglyoxal(MG)-induced injury of hippocam-pal neurons. Methods Primary cultured of hippocampal neurons from 1-day-old Sprague-Dawley rats were incuba-ted with MG for different time and dose period. Cells proliferation were assayed by methyl thiazolyl tetrazolium (MTT),and apoptosis was quantified by flow cytometer using annexin V-FITC and propidium iodide (PI) stai-ning. The protein and mRNA levels of brain-derived neurotrophie factor (BDNF) and tyrosine kinase B(TrkB) were assayed with Western Blotting and real-time PCR. Results Treatment with MG resulted in a concentration-dependent (r=0.946, P < 0.01) and time-dependent (r=0.993, P < 0.01) decreasing neurons viability. Com-pared with Oh group(1. 633±0. 153)%, 100 μM MG treatment for 2h,6h, 12h and 24h,the cellular apeptosis rate were significantly increased ((2. 833±0. 153)%, (3. 367±0. 153)%, (4. 433±0. 404)% and (8. 833± 0. 306)% respectivdy,all P<0.01). MG also increased the BDNF mRNA and protein expression after 12h treat-ment (P<0.05 or P<0.01),but decreased the TrkB mRNA and protein expression in the cells after 6h treatment (P<0.05 or P < 0.01). Conclusion MG has direct toxic effect on hippocampal neurons and can impaire the BD-NF-TrkB signal pathway by inhibiting the expression of TrkB,and increasing the apoptosis of hippocampal neurons. Key words: Neuron; Methylglyoxal; Brain-derived neurotrophic factor; Receptor,TrkB
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Objective To investigate the mechanisms of methylglyoxal(MG)-induced injury of hippocam-pal neurons. Methods Primary cultured of hippocampal neurons from 1-day-old Sprague-Dawley rats were incuba-ted with MG for different time and dose period. Cells proliferation were assayed by methyl thiazolyl tetrazolium (MTT),and apoptosis was quantified by flow cytometer using annexin V-FITC and propidium iodide (PI) stai-ning. The protein and mRNA levels of brain-derived neurotrophie factor (BDNF) and tyrosine kinase B(TrkB) were assayed with Western Blotting and real-time PCR. Results Treatment with MG resulted in a concentration-dependent (r=0.946, P < 0.01) and time-dependent (r=0.993, P < 0.01) decreasing neurons viability. Com-pared with Oh group(1. 633±0. 153)%, 100 μM MG treatment for 2h,6h, 12h and 24h,the cellular apeptosis rate were significantly increased ((2. 833±0. 153)%, (3. 367±0. 153)%, (4. 433±0. 404)% and (8. 833± 0. 306)% respectivdy,all P<0.01). MG also increased the BDNF mRNA and protein expression after 12h treat-ment (P<0.05 or P<0.01),but decreased the TrkB mRNA and protein expression in the cells after 6h treatment (P<0.05 or P < 0.01). Conclusion MG has direct toxic effect on hippocampal neurons and can impaire the BD-NF-TrkB signal pathway by inhibiting the expression of TrkB,and increasing the apoptosis of hippocampal neurons. Key words: Neuron; Methylglyoxal; Brain-derived neurotrophic factor; Receptor,TrkB
Key concepts: Tropomyosin receptor kinase B, Methylglyoxal, Hippocampal formation, Internal medicine, Propidium iodide, Brain-derived neurotrophic factor, Endocrinology, Neurotrophic factors