ABSTRACT
BACKGROUND:Neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine can induce the clinical, biochemical and pathological characteristics similar to those observed in primary Parkinson’s disease. OBJECTIVE:To observe the effects of repetitive transcranial magnetic stimulation on the proliferation of endogenous neural stem cells of Parkinson’s disease model mice and the mood change. METHODS:A total of 72 male C57BL/6J mice were randomly divided into four groups:normal saline group, Parkinson’s disease model group (model group), sham-repetitive transcranial magnetic stimulation group (sham group) and repetitive transcranial magnetic stimulation group. The mice received 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine injection×4 to establish acute Parkinson’s disease models. The mice in the normal saline group were injected the same volume saline. And 24 hours after the last injection of 1-methyl-4-phenyl-1,2,3, 6-tetrahydropyridine, the mice in the repetitive transcranial magnetic stimulation group received five trains of repetitive transcranial magnetic stimulation, 1 Hz for 25 seconds, at an intensity of 1 Tesla daily for 1, 3, 7 consecutive days. Sham group mice were not exposed to the magnetic field. No treatment was performed in the mice of model group. The mood change was evaluated using the elevated-plus maze testing before and after repetitive transcranial magnetic stimulation treatment. The change in expression of nestin in the subventricular zone was observed by using immunohistochemical technique. RESULTS AND CONCLUSION:(1) Elevated-plus maze testing:There was no statistical significance about percentage of opening arm time accounting for total time among groups and at different time points in each group, but after stimulation, the percentage of opening arm time accounting for total time showed a declined tendency. (2) The results of nestin immunohistochemical staining:Compared to the normal saline group, the number of nestin-positive cells of the model group was increased at days 3 and 7, and there was no statistical significance in the number of nestin-positive cells between model group and sham group;Compared to the sham group and model group, the number of nestin-positive cells of repetitive transcranial magnetic stimulation group were evidently increased;The proliferation of endogenous neural stem cells was time-dependent, endogenous neural stem cells exhibited outward migration gradual y along the certain way, and some cells were able to migrate to the corpus cal osum at day 3, and even to the cerebral cortex at day 7. Repetitive transcranial magnetic stimulation can promote the endogenous neural stem cells in a time-depended manner.
ABSTRACT
Objective Using shotgun mass spectrometry to detect proteins probably contained in bone marrow stromal cells(BMSCs) conditioned medium. MethodsMixed with BMSCs conditioned medium was divided into two parts which is(>5kD and <5kD) by means of ultrafiltration. The two parts were used to culture neural stem cells(NSCs) separately, and the proportions of neurons, astrocytes and oligodendrocytes in the offsprings of NSCs were calculated, then the effective part that could regulate the differentiation of NSCs was detected by Shotgun mass spectrometry. Results The BMSCs conditioned medium which is >5kD could promote the NSCs differentiate into more neurons and oligodendrocytes. The SDS-PAGE of this part showed that the most proteins were above 14kD, then the protein bands were enzymed. In total, 456 proteins were identified by Shotgun mass spectrometry after all the protein bands were enzymed, there were 154 similar proteins, 17 hypothetical proteins and 56 unknown proteins. And in the rest of 229 proteins, most of them were cytoskeletal proteins, secreted proteins, signal transduction proteins, enzymes, transporter and so on. Conclusion Many proteins secreted by BMSCs could regulate the differentiation of NSCs so as to prove the protein components probably existed in the BMSCs conditioned medium.