| Objective:To establish the damaged model of superior cervical sympathetic ganglion(SCSG)which were transplanted by induced neuron stem cells from bone marrow, observe therecorvery of pupil, detect the variation of microtubule associated protein2(MAP-2) andglial fibrillary acidic protein (GFAP) in SCSG. and discuss the recovery effect and pathwaywith neuron stem cells to nerve injury.Methods:Got the bone marrow from the white New-Zealand rabbits,femur about one week old,cultivated the rabbit bone marrow mesenchymal stem cells (BMSCs) with the method ofdensity gradient centrifugation and adherent-culture, induced the BMSCs into neuron stemcells with the fibroblast growth factor (bFGF) and the ciliary neurotrophic factor (CNTF),authenticated the phenotypic characteristic of BMSCs and neuron stem cells.Squeezed the rabbit,s SCSG with straight needle forceps about10seconds each time,prepared for the rabbit damaged model of SCSG (24), divided into three grouprandomly:the model control group (group A), the nutrient solution control group (group B),the intervented group with neuron stem cells (group C).7days after the rabbit SCSG damaged successfully, injected1milliliter normal salineinto the rabbits,era-edge vein of group A with a syringe, the same method,1milliliternutrient solution without neuron stem cells into the rabbits,era-edge vein of group B,1milliliter nutrient solution within2×106neuron stem cells into the rabbits,era-edge veinof group C. Observed the variation of the damaged side pupil diameter at the1st,7th,10th,14th,21th day after transplantation, killed the rabbits after transplanted21days, anddetected the variation of MAP-2, GFAP with the immunohistochemical method in thedamaged SCSG. Results:1: After the rabbit SCSG was damaged, the same side eye showed Horner syndrome:miosis, palpebral fissure narrowed, enophthalmos. The Horner syndrome was obvious afterthe SCSG was damaged28days.2:The CD44and CD54, which were used to detecte the phenotypic characteristic of theBMSCs with the method of immunocytochemistry, were positive, and the CD34wasnegative，all these were conform to the phenotypic characteristic of the BMSCs; the NSEand Nestin, which were used to detecte the phenotypic characteristic of the neuron stemcells with the method of immunocytochemistry, were positive, and these were conform tothe phenotypic characteristic of the neuron stem cells.3. The pupil diameters (millimeter) in the side of damaged SCSG, at the1st,7th,10th,14th,21th day after the neuron stem cells were transplanted, in group A were7.92+0.38,8.02+0.30,8.13+0.37,7.96+0.36,8.12+0.25；in group B were5.82+0.42,5.84+0.34,5.68+0.29,5.91+0.31,5.80+0.36； in group C were5.85+0.32,5.75+0.32,5.64+0.37,5.84+0.27,5.59+0.23; in group D were5.64+0.33,5.72+0.35,6.79+0.33,7.98+0.28,8.06+0.21; the measurement of pupil diameter between group B and C was nostatistically significant at each time point (P>0.05)，but lower than group A; and themeasurement of pupil diameter between group A and D was statistically significant at the1st,7th,10th day, but no statistically significant at the14th、21th day; Compared withgroup B and C, the measurement of pupil diameter in group D was no statisticallysignificant at the1st,7th days after the neuron stem cells were transplanted, butstatistically significant at the10th,14th、21th day（P<0.05）; the compare results whineach group: there were no statistically significant at each time in group A, group B andgroup C (P>0.05). In group D, the difference of the pupil diameter was no statisticallysignificant between at the1st day and the7th day(P>0.05); Compared with the1st,7th day,the difference at the10th day was statistically significant（P<0.05）, and the difference atthe14th day was statistically significant compared with the1st,7th10th day（P<0.05）, thedifference at the21th day was statistically significant compared with the1st,7th10th day（P<0.05）, but no statistically significant compared with the14th day(P>0.05). 4.pathological tissue morphology:(1)Rabbit superior cervical sympathetic ganglion HE staining:neuron cells in group A andgroup B were rare.We find that they arranged sparsely, were uneven in morphology andsize, had more intercellar substance and less new blood vessels,even cell nucleus exhibitedpyknosis;In contrast, in group C neuron cells growed more closely and arranged moreorderly,had more consistent shape and size,richer new blood vessels,fewer intercellarsubstance,cell nucleus without pyknotic.(2)MAP-2:The results of immunohistochemistry showed that MAP-2in group A and groupB were negative expression, but it were positive in group C. The measurement of averageoptical density value in group A, B, C, D were0.787+0.245,0.457+0.022,0.447+0.020,0.771+0.027. There are no marked difference between group A and group D, betweengroup B and group C (P<0.05), but compared with group A and D, the differences in groupB and group C were statistically significant (P<0.05).(3)GFAP:The results of immunohistochemistry showed that GFAP in group A and group Bwere negative expression, but it were positive in group C. the measurement of averageoptical density value in group A, B, C, D were0.249+0.020,0.640+0.038,0.658+0.031,0.271+0.022respectively, there were not statistically difference between group A andgroup D, between group B and group C (P>0.05). while the value in group A and D waslower than the other two groups, and there difference was statistically significant (P <0.05).Conclusions:1. The marker protein of neuron (MAP-2) is degressive, and the marker protein ofspongiocyte (GFAP) is increased in the damaged SCSG.2. The neuron stem cells can recover the the neurological function of the damaged SCSGin rabbit, the mechanism may be through promoting the expression of MAP-2,andinhibiting the expression of GFAP. |
PDF Full Text Request