| Objective: Motor neuron disease (motor neuron disease, MND) is agroup of unknown etiology selective infringement spinal horn cells, brainstemmotor neurons, cortical pyramidal cells and pyramidal tract of chronicprogressive neurodegenerative disease. Amyotrophic lateral sclerosis(amyotrophic lateral sclerosis, ALS) is the highest incidence type of motorneuron disease, is a selective involvement of the spinal anterior horn cells andbrainstem motor nuclei of the brain’s motor cortex progressive fatalneurological degeneration, simultaneously involving the upper and lowermotor neurons characterized[1-2]. Patients with first appearance of symptomsmore than3to5years after the due to progressive muscle weakness andatrophy, respiratory failure and death. Currently, the clinical in addition toriluzole can limited slow the course of disease, there is no effective treatment.[3]such as stem cell transplantation, gene therapy new methods is still beingstudied, familial ALS accounts for5to10%,20%of which with Cu/Znsuperoxide dismutase (SODl) gene mutation[4-6].Nerve growth factor (nerve growth factor, NGF) was first discovered bythe Levi Montalcini sarcoma cells in mice. NGF has maintained neural stemcells of the survival, neural stem cell proliferation, differentiation andmigration, inhibition of apoptosis, nutrition neurons, protection and repair ofdamaged nerve effects.[7]NGF can promote the in vitro neural precursor cellsdifferentiation of into immature neurons and astrocytes. NGF can promoteneural stem cell survival, proliferation, differentiation and directed migrationrole, helping the central nervous system regeneration and repair of nerves afterinjury, and brings hope for the MND treatment. the liver have first pass effectand blood-brain barrier (Blood-brain barrier BBB) barrier, the classic route ofintravenous and subcutaneous administration, effects of the drug did not make the play to the greatest degree. SOD1-G93A transgenic mice areinternationally recognized, representative of an animal model of familialALS. Therefore, the experimental study using nasal SOD1mice treated withdifferent concentrations of NGF observed brain hippocampus dentate gyrusBrdU, Nestin the number of ways of expression and neuronal, evaluationwithin their SOD1transgenic mice derived neural stem cells the proliferationof regulation, aimed at finding a way to bypass the BBB and make NGF in thecentral target site quickly reach new route of administration, the best effectconcentration.Methods:1experimental model and groups:100days of non-transgenic femalemice6s normal control group, divided into group A; internationallyrecognized100days (symptoms of) SOD1-G93A transgenic female mice36(by the Chinese Academy of Medical Science Experiment animal ResearchCenter). Were randomly divided into control group (B)6s; nasaladministration group (C)30s. Then in accordance with differentconcentrations of NGF nasal administration, nasal administration group willbe divided into:15mg/kg dose group (C1);20mg/kg dose group (C2),25mg/kg dose group (C3),30mg/kg dose group (C4),35mg/kg dose group C5.2routes of administration and methods:. A group and B group nasalsaline, group C nasal give NGF, each dose given5ul, interval2minadministration again, according to the total amount of each group were givenA:15ul, B:15ul, C1:15ul, C2:20ul, C3:25ul, C4:30ul, C5:35ul. Using thesame method of continuous administration of the drug for3days.3specimens of each group were randomly selected half by SABCimmunohistochemical staining method to display the endogenous neural stemcells within the brain and cell proliferation. Image analysis and counting thenumber of positive cells Brdu Nestin and analyze changes in brain nerve cellproliferation compared between groups A, B, C group and endogenous neuralstem cell proliferation and differentiation.4The other half of the specimens intraperitoneal injection of sodium pentobarbital (40mg/Kg) after the mice were anesthetized, decapitated, braintissue isolated from the hippocampus, using Western-Blot assay proteinexpression levels.Results:1HE and Masson staining showed that: A group of BrdU-positive cells inthe hippocampal dentate less back to exist. B Group cortical had more Brduand Nestin positive cells, while bilateral hippocampal dentate gyrus regionalso has Nestin positive cells, but the region Brdu positive cells the relativelysmall number; But C1-C5group Nestin positive cells and Brdu weresignificantly increased (P <0.05); when the NGF concentration of25mg/kg,with concentration increasing, the number of Nestin and Brdu positive cellsproliferation proliferation obvious than before.2Western Blot Changes related protein gene: Western blot analysisshowed: normal control group, and the brain has Nestin Brdu certain level ofprotein expression; Group B mice relative to the control group of micehippocampus dentate gyrus and Brdu Nestin protein higher expression. GroupC along with the gradual increase in nasal dose, Nestin and Brdu protein alsowill be gradually increased, but the C4, C5group compared weakened C3protein group, the difference was significant (P <0.01).Conclusion:1Symptoms of SOD1transgenic mouse brain there endogenous neuralstem cell proliferation.2NGF effect on motor neuron disease in mice and its concentration. Lowconcentrations (<15mg/kg), Mainly for the protective effect on neurons, butno significant inhibition of the proliferation of NSCs， At a concentration of25mg/kg, you can significantly inhibit the proliferation of endogenous neuralstem cells; When the concentration of30mg/kg or more of neural stem cells invitro, with the increasing concentration showed an increasing trend; Whenconcentrations exceed35mg/kg when can produce significant adversesystemic reactions. |
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