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Effects of NGF on the Neural Cells and Egr-1 Gene Expression in Patients with Epilepsy

Author: FangQiong
Tutor: ChenYanHui
School: Fujian Medical
Course: Pediatrics
Keywords: Epilepsy Nerve Growth Factor Early growth response gene 1 Hippocampal Nerve cells People
CLC: R742.1
Type: Master's thesis
Year: 2008
Downloads: 68
Quote: 0
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Abstract


[Objective] To understand the nerve growth factor in the cerebrospinal fluid of patients with epilepsy (nerve growth factor, NGF) level of different concentrations of NGF on human epileptic hippocampal CA1 region of the total number of nerve cells, astrocytes, the number of neuronal survival impact different concentrations of NGF of nerve cells Egr-1mRNA expression to explore NGF on the role and mechanisms of epileptic brain damage nerve cells. [Methods] ELISA (enzyme-linked immunosorbent assay, ELISA) were measured epilepsy group (n = 28) and non-epileptic control group (n = 21) in cerebrospinal fluid NGF levels; using dissociation cell instantly culture techniques people with epilepsy epileptiform discharges after inspection originated in the temporal lobe or hippocampus and pathological examination found no specific pathological changes in the nerve cells of the hippocampal CA1 (n = 16) were cultured, and the establishment of concentration were 12.5, 50, 100 ng / ml the NGF plus drug group and without NGF blank control group, after 4h incubation, nuclear staining by immunofluorescence cytochemistry double benzene imide (Bisbenzimide, Bb), joint cell-specific marker proteins GFAP, MAP2 mark Star shaped glial cells, neurons, and count the total number of nerve cells survive, and astrocytes and neurons number and calculation of astrocytes and nerve elements of the total percentage of nerve cells, while taking advantage of the RT-PCR technique half different concentrations of NGF quantitative analysis of the impact of early growth response gene -1 (early growth response gene-1, Egr-1) mRNA expression in nerve cells. [Results] 1. Cerebrospinal fluid in NGF content epilepsy group and non-epileptic control group, respectively (27.31 ± 6.36) pg / ml, (24.09 ± 4.96) pg / ml, NGF levels in the cerebrospinal fluid groups no significant difference (P gt; 0.05 ). 2 in NGF concentration 12.5,50,100 ng / ml, the number of nerve cells in the hippocampal CA1 region were (82.02 ± 11.08), (104.71 ± 14.24), (127.11 ± 14.35) / field of view, and without NGF blank The control group (67.71 ± 9.21) / field compared, the difference was statistically significant (P lt; 0.01), total number of neuronal survival with NGF concentration increased and increased with the NGF dose was the dose-effect relationship (P lt; 0.01). NGF concentration 0,12.5,50,100 ng / ml, the number of astrocyte GFAP () were (23.11 ± 4.39), (30.73 ± 5.16), (44.25 ± 6.76), (59.57 ± 6.23) / vision; MAP (2) the number of neurons, respectively (8.82 ± 1.74), (14.81 ± 2.87), (22.51 ± 3.10), (32.17 ± 2.52) / vision, astrocytes and nerve neuron survival number of NGF-dose dose-effect relationship, with increased NGF concentration increased (P lt; 0.05). With the NGF concentration increased astrocytes and neurons accounts for the percentage of nerve cells was increased, showing the same trend of increasing. The NGF concentration 0,12.5,50,100 ng / ml, GFAP () astrocytes and MAP2 () neurons representing the percentage of nerve cells in the median (interquartile range) 34.94 (31.16 to 36.91)%, 38.49 (34.70 to 39.94)%, 42.36 (40.22 to 45.13)%, 45.92 (43.79 to 51.53)% and 13.01 (10.65 to 15.55)%, 18.35 (16.37 to 19.92)%, 21.94 (21.36 to 22.84) 25.33 (23.98 to 26.84)%; comparing different concentrations of NGF GFAP () astrocytes and MAP2 () neurons account for the impact of nerve cells percentage change found no significant difference significance (P gt; 0.05). When NGF concentration, as 0,12.5,50,100 ng / ml, the nerve cell expression of Egr-1mRNA AEgr-1/Aβ-actin (0.46 ± 0.14), (0.63 ± 0.14), (0.80 ± 0.15) , (1.12 ± 0.12), the treatment group were not added NGF control group comparison, the existence of significant forward difference (P lt; 0.01), the same nerve cells in the expression of Egr-1mRNA amount with NGF dose showed a dose-response relationship, with the NGF concentration increased and expression was increased (P lt; 0.01). And in the NGF effect, the number of nerve cells and the corresponding expression of Egr-1mRNA the amount of a positive correlation (r = 0.780, P lt; 0.01). [Conclusion] The results showed that refractory epilepsy in cerebrospinal fluid NGF levels than non-epileptic control group no significant increase; NGF can improve in vitro cultured human epileptic brain hippocampal CA1 (12.5 to 100) ng / ml range the number of cells and GFAP () astrocytes and MAP2 () the number of neurons, the role of NGF concentration increased was positively correlated, suggesting that NGF has a protective effect on nerve cells; comparing different concentrations of NGF star enhance the role of glial cells and neurons survival showed no significant differences, suggesting that NGF promote astrocytes and neurons in hippocampal CA1 region of people with epilepsy have a protective effect on nerve cells; NGF expression of Egr-1mRNA role, the increase in the amount of the expression of NGF concentration was a positive dose-effect relationship, suggesting that Egr-1 may play an important role in the NGF-mediated nerve cell protection.

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CLC: > Medicine, health > Neurology and psychiatry > Neurology > Brain diseases > Epilepsy
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