The Therapeutic Effects Of Co-transplantation For Treating Cerebral Ischemic In Rats Through Different Paths

OBJECTIVETo observe the rat neurological function recovery and explore the protective mechanism and correct transplanting path which were respectively through stereotaxis, vena caudalis and carotid artery path treated middle cerebral artery occlusion(MCAO) model by mesenchymal stem cells(MSCs) and endothelial progenitor cells(EPCs).METHODSMSCs and EPCs were respectively isolated, proliferated, purified and passaged from the long bone of 100g SD rats.MACO model was established by improved method.Forty MCAO rats were divided into BPS control group (A), vena caudalis group (B), carotid artery group(C) and stereotaxis group(D) at random. 2×10~6MSCs and 2×10~6EPCs were injected into MACO model rats in group B, C and D through vena caudalis, carotid artery and tereotaxis paths respectively and 10μl BPS fluid transplanted into MACO model rats in A group at 24-hour after made modes. Bederson neurological score, cerebral infarction volume, HE dyeing, brain-derived neurotrophic factor(BDNF), Vascular endothelial growth factor (VEGF) were investigated after 14 days.RESULTS1 The volume of cerebral infarction: The infarction volumes of group A, B, C and D were 216.3+4.86mm3, 206.0+5.33mm3, 198.2+4.19mm3 and 191.2+4.96mm3 respectively. The infarction volumes between each two groups were statistically significant(P<0.05) and they in group D was the smallest.2 The scores of Nerve behavior: The Bederson neurological score of group A, B, C and D were 3.4+0.67, 3.1+0.77, 3.4+0.52 and 3.3+0.67 respectively and not statistically differences(P>0.05),While They were 3.0+0.82, 2.4+0.84,2.0+0.67 and 1.2+0.42 respectively in the 14-day after transplanted. The scores at each two groups were statistically significant except between group A and group B, group B and group C, (P<0.05). Neurological function was recovered bestly in group D. 3 Pathological morphology(1) BDNF expressions: The positive expressions of BDNF in group A were less than those in group B, group C and group D. The cells with positive expressions of BDNF almost were pyramidal cells or granulosa cells at hippocampus and cortex, and their cytoplasm were tan and nuclei was not stained. The positive expressions of BDNF between each two groups were statistically significant(P<0.05) and they in group D was the highest.(2) VEGF expressions: The numbers of VEGF positive cells in cerebral penumbra of group A were 10.50+1.96 which is the lowest and less than those in group B, group C and group D. The numbers of VEGF positive cells between each two groups were statistically significant(P<0.05). They were 26.4+1.90 in group D and the highest.(3) HE staining: The cortex nervous cells in group D which were rich,abundant cytoplasm and damaged slighter than those in group A, B and C were relatively well except some small softening lesions at low power microscope. The cells in group A damaged most seriously than those in other groups. The cells in group B which structure were bad and edema damaged more seriously than those in group C.CONCLUSION1 The mechanisms of transplantation MSCs with EPCs treating ischemic brain injury were upgrading secretion of BDNF,VEGF in brain, promoting new nerve cells neogeneses and angiogenesis and increasing blood flow to ischemic areas to improve nerve functions.2 The stereotaxis injection was the best method which was more effective than vena caudalis and carotid artery path. The carotid artery was better than vena caudalis path.