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目的探讨脐血单个核细胞(umbilical cord blood monocytes,UCBMC)移植对低龄大鼠脑白质损伤(white matter damage,WMD)中少突胶质细胞及髓鞘碱性蛋白表达的影响。方法将3日龄健康Sprague-Dawley新生大鼠80只随机分为正常对照(control,CON)组、WMD组、WMD+UCBMC组与UCBMC对照组。通过单侧缺血联合缺氧的方法制成WMD模型(每个时间点每组各5只),造模24h后,CON组和WMD组经侧脑室注入2μL PBS,UCBMC对照组和WMD+UCBMC组经侧脑室注入3×106 UCBMC。移植后24h、3d、7d和14d,采用2’,3’-环核苷酸3’-磷酸二酯酶(CNPase)/活性半胖氨酸蛋白水解酶(Cleaved Caspase-3)及髓鞘碱性蛋白(MBP)/4’6-二脒基-2苯基吲哚(4’6-diamidino-2-phenylindole,DAPI)免疫荧光双标观察UCBMC移植对纹状体处少突胶质细胞及MBP的变化,并行机制的探讨。结果移植后24h、3d和7d,WMD组CNPase+Cleaved Caspase-3+细胞数均高于CON组、WMD+UCBMC组与UCBMC对照组(均P<0.01),且在24h开始增加,3d达高峰,7d有所下降;WMD+UCBMC组CNPase+Cleaved Caspase-3+细胞数显著少于WMD组(P<0.01)。移植14d后,WMD组MBP+DAPI+细胞数显著少于CON组及UCBMC对照组(P<0.01);WMD+UCBMC组MBP+DAPI+细胞数显著高于WMD组,仍少于CON组(P<0.01)。结论 UCBMC侧脑室移植可减少脑白质损伤低龄大鼠的少突胶质细胞的凋亡,减少髓鞘的脱失,从而修复损伤的脑白质。
Objective To investigate the effects of umbilical cord blood mononuclear cells (UCBMC) transplantation on the expression of oligodendrocyte and myelin basic protein in young white matter damage (WMD) rats. Methods Eighty three-day-old healthy Sprague-Dawley neonate rats were randomly divided into control group (CON), WMD group, WMD + UCBMC group and UCBMC control group. WMD model was established by unilateral ischemia combined with hypoxia (5 rats in each group at each time point). After modeling for 24 h, 2 μL PBS, UCBMC control group and WMD + UCBMC Group by the lateral ventricle injection of 3 × 106 UCBMC. After 24 h, 3 d, 7 d and 14 d after transplantation, 2 ’, 3’-cyclic nucleotide 3’-phosphodiesterase (CNPase) / active proteasome protease (Cleaved Caspase-3) and myelin Immunofluorescence double labeling of MBP / 4’6-diamidino-2-phenylindole (DAPI) to observe the effect of UCBMC transplantation on oligodendrocyte and MBP changes, the discussion of parallel mechanism. Results The numbers of CNPase + Cleaved Caspase-3 + cells in WMD group were significantly higher than CON group, WMD + UCBMC group and UCBMC group (all P <0.01) 24h, 3d and 7d after transplantation, (P <0.01). The number of CNPase + Cleaved Caspase-3 + cells in WMD + UCBMC group was significantly less than that in WMD group (P <0.01). The number of MBP + DAPI + cells in WMD group was significantly less than that in CON group and UCBMC control group (P <0.01) after 14 days of transplantation. The number of MBP + DAPI + cells in WMD + UCBMC group was significantly higher than that in WMD group ). Conclusions UCBMC can reduce the apoptosis of oligodendrocytes in young rats with white matter lesion, reduce the loss of myelin, and repair damaged white matter.