Myelin Basic Protein Active Immunization Of Hemorrhagic Stroke In Rats Study The Protective Effect

Objective: myelin basic protein (myelin basic protein, MBP) active immunization of hemorrhagic stroke in rats, behavioral observation, and immunohistochemical study, GFAP and NF200 of hematoma in the brain tissue around the expression, and application of electron microscopy hematoma in the brain tissue around the ultrastructural changes, the study of hemorrhagic stroke MBP secondary brain damage in the neuroprotective effect.Methods: healthy adult female SD rats, using autologous blood injection method of rat cerebral hemorrhage model after 12 hours, behavioral score to 2 minutes, as the inclusion criteria. Selected 84 rats by using random number table is divided into MBP, egg albumin (Ovalbumin, VOA), PBS and four blank control group (n=21). With incomplete adjuvant IFA Fook-emulsification of the MBP (300μg /), VOA (200μg /), PBS (200μg /) three groups were immune SD rats, the control group only cerebral hemorrhage model, 1,2,3,5,7,14,21 days after vaccination, a behavioral score, and in the days after immunization 1,3,5,7,14,21 animals were killed, brain tissue to glial fibrillary acidic protein (Glial fibrillary acid protein, GFAP) and Neurofilament protein 200 (Neurofilament protein, and NF200) immunohistochemical staining. In the first seven days after immunization of each group will take three electron microscope.Results:⑴behavioral score: MBP immune rats immunized with 1 to 3 days behavioral score was superior to VOA, PBS immune group and the control group (P <0.05);⑵GFAP immunohistochemistry staining of observation: cerebral hemorrhage after immunization 1 ~ 5 days, MBP Group hematoma around GFAP positive cells and VOA, PBS and the control group there were significant differences (P <0.05). After three groups showed no statistical significance (P> 0.05); cerebral hemorrhage after immunization 7 ~ 21 days, the groups of GFAP positive cells showed no significant difference (P> 0.05).⑶NF200 immunohistochemical staining of observation: MBP Group NF200 positive cells in the immune after 3 to 14 days after the three groups compared with a significant difference (P <0.05), after the three groups showed no significant difference (P>0.05).⑷TEM observation: cerebral hemorrhage in the first seven days after immunization, MBP immunohistochemical neuronal cells and myelinated nerve fibers in the cell-rich, Neurofilament neat and found no vascular endothelial cells edema. The other three groups neuronal cells and nerve fibers of the deformation of cells or disappeared, Neurofilament are disordered, and that vascular endothelial cell edema.Conclusion: (1) to promote active immunization MBP and NF200 GFAP expression;(2)MBP immunohistochemical behavioral observation and hematoma surrounding ultrastructural changes clearly better than the other group. MBP immune therapy can block hemorrhagic stroke secondary brain damage, thereby promoting the improvement of neurological symptoms, or hematoma surrounding neurons recovery;(3) MBP own initiative immune activation effect of T cells through the blood-brain barrier, gathered around the hematoma in improving the micro-environment of the neurons, blocked cerebral hemorrhage secondary brain damage and the recovery of neuronal function play an important role.Objective: To study the rats hemorrhagic stroke, immune therapy on the MBP BDNFmRNA and protein expression. MBP study in the rat central nervous system injury confrontation secondary brain damage - that is, autoimmune nerve protection mechanism.Methods:⑴healthy adult female SD rats, using autologous blood injection method of rat cerebral hemorrhage model after 12 hours, behavioral score, in the light Longa method developed standard score to 2 minutes, as included in the pilot standards. 72 selected by random number table is divided into MBP, ovalbumin VOA, PBS and four blank control group (n = 18). With incomplete adjuvant Fook-emulsification of the MBP (300μg /), VOA (200μg /), PBS (200μg /) three groups were immune SD rats after vaccination, 1,2,3,5,7, 14, 21 days behavioral score,In the days after immunization 1,3,5,7,14,21 animals were killed, brain tissue to the brain-derived neurotrophic factor (brain derived neurotrophi factor, BDNF) immunohistochemical staining.⑵Preparation of 36 with SD rats bleeding were randomly divided into MBP group and the control group (N = 18). 12 h after cerebral hemorrhage emulsified with the IFA MBP (300μg /) MBP immune rats. 1,3,5,7,14,21 days after the bleeding by RT-PCR detection of the two groups BDNFmRNA hematoma in the brain tissue around the expression.Results:(1)behavioral observation: MBP immunohist- ochemical rat neural score was superior to VOA, PBS immune group and the control group;⑵BDNF Immunohistochemical detection; BDNF mainly by microglial cells, one day after immunization, MBP Group BDNF positive cells with the VOA group, PBS group, the control group compared with a significant difference (P <0.01) Three days after immunization MBP Group BDNF positive cells compared with the three groups after a significant difference (P <0.05) between the three groups after showed no statistical significance (P> 0.05); Immunity after 5 to 21 days, four groups of rats around the hematoma BDNF positive cells further reduced, the group there was no significant difference (P> 0.05). (3)BDNFmRNA RT-PCR results show that: MBP group at different time points BDNFmRNA hematoma in the brain tissue around the expression of the control group, all time points compared between the two groups were significantly different (P <0.05) and, in particular 5,7 days after immunization between the two groups significant difference (P <0.01). MBP Group 1,14 days in the first two peaks, one day after immunization secretion BDNFmRNA highest, seven days after immunization minimum, 14 days once again reached a peak.Conclusion: (1) MBP rat brain hemorrhage after active immunization can BDNFmRNA increases BDNF protein and the expression of neuronal improve micro-environment, and promote neuronal survival and functional recovery;(2) MBP treatment by activating their immune effector T cells gathered around the hematoma, central nervous system injury after blocking secondary brain damage occurred, and promote restoration of the central nervous system.