| Intracerebral hemorrhage and other central nervous system(CNS) damage often could not acqired complete recovery, which is closely related to the difficulty of axonal regeneration in CNS. Effect of conventional therapies is considerable limited.Stem cells bring hope for the treatment of central nervous system damage on their unique characteristics of promoting the regeneration of injured tissue and cells. Among them, Bone marrow-derived Mesenchymal Stem Cells(BMSCs) offer significant practical advantages over other types of stem cells since they can be obtained from the adult BM, applied without ethical and immune rejection obstacles, easily cultured and expanded in vitro under GMP conditions displaying a very low risk of malignant transformation and have the potential to differentiate into cells of the multiple lineages.Myelin-associated glycoprotein(MAG), Oligodendrocyte myelin glycoprotein(OMgp), glycoproteins mainly expressed in myelin sheath of central nervous system, both are identified as potent myelin-associated inhibitory factors of axonal regeneration. The studies of the two inhibitory factors indicate a new therapeutic target of neural regeneration. Whether transplanted BMSCs mediate neuroregeneration by regulating the expression of these myelin-associated inhibitory factors needs further study.In this study, we established rat intracerebral hemorrhage(ICH) model by collagenase inducing method, compared neurological function scores and expression levels of MAG, OMgp in perihematoma tissue among sham-operated group, ICH group, BMSC group at different time points after surgery to investigate the effect of intravenous administration of BMSCs on the improvement of functional deficits and the expression of MAG and OMgp in perihematoma tissue in rats with ICH, which would be expected to further verify the inhibitory effect of BMSCs on myelin-associated inhibitory factors and to provide more theoretical basis and experimental support for the clinical application of BMSCs. Objectives:To investigate the effect of intravenous administration of bone marrow-derived mesenchymal stem cells (BMSCs) on the improvement of functional deficits and the expression of MAG and OMgp in perihematoma tissue in rats with intracerebral hemorrhage.Methods:1.Rat BMSCs were isolated and cultured using the whole bone marrow adherent method, purified by repeated passages, identified by flow cytometry detecting the expression levels of cell surface markers. Took the 5th to 8th generation cells for experiment. Before transplantation, BMSCs were labeled with BrdU.2.Stereotaxically injected collagenase IV into striatumto establish rat ICH model. Sprague-Dawley(SD) rats(weights250-300g) were randomly divided into sham-operated group(sham group), intracerebral hemorrhage group(ICH group) and intracerebral hemorrhage plus BMSCs intervention group(BMSC group). Rats in sham group only received stereotaxic puncture into striatum, no collagenase injection. Rats in BMSC group were transfused with BMSCs (1×107/rat) via tail vein 24 hours after operation.3.On day 1,3,7,14 after operation, neurological function tests(mNSS and MLPT) were performed among all three groups and animals were sacrificed at the corresponding time points for detecting the expression of MAG, OMgp by immunohistochemical method, immunofluorescent staining of BrdU was performed to observe the distribution of the BMSCs in brain.Results:1. No obvious neurological function deficit was seen in sham group on day 1,3,7,14 after operation. In ICH group, the degree of neurological function deficit reached the peak on day 1 after operation, lasted to day 3, on day 7, recovery could be observed, and on day 14 much more recovery could be seen. Compare to ICH group, BMSC group had significantly lower mean neurological function rating scores on day 3,7,14 after operation(P< 0.05).2. The mean expression levels of MAG, OMgp in neurons and gliocytes in perihematoma tissue in ICH and BMSC group were both significantly higher than that in sham group on day1,3,7,14 after operation, meanwhile BMSC group had significantly lower mean expression levels than ICH group on day 3,7,14 after operation(P<0.05).3. In BMSC group, Brdu positive BMSCs by Immunofluorescence staining could be detected under fluorescence microscope, and most of these cells were surrounding the haematoma, only very few scattered Brdu positive cells could be seen in the opposite brain area.Conclusions:1. Intravenous administration of BMSCs significantly improved functional deficits in rats with experimental ICH.2. There were up-regulation of MAG, OMgp in perihematoma tissue.3. Transplantation of BMSCs could significantly promote the neurological function recovery of rat ICH model, which might be associated with its down-regulation of MAG, OMgp in perihematoma tissue. |
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