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24 h 后，CON组和WMD组经侧脑室注入2 μL PBS，UCBMC 对照组和WMD+UCBMC 组经侧脑室注入3×106UCBMC。移植后24 h、3 d、7 d和14 d，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24 h、3 d和7 d，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24 h开始增加，3 d达高峰，7 d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14 d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。
Abstract
Objective To explore the effects of umbilical cord blood monocytes (UCBMC) transplantation on oligodendrocytes and MBP protein in White Matter Damage(WMD) neonatal rats. Methods Eighty three-day-old Sprague-Dawley rats were randomly divided into nomal control (CON),WMD,WMD+UCBMC and UCBMC groups.The WMD model was made by using the method of unilateral ischemia joint with hypoxia (five rats at each time point in each group),twenty-four hours after hypoxia ischemia,the rats of the CON and WMD groups were injected with 2 μL phosphate buffered saline (PBS ),and the UCBMC and WMD+UCBMC groups were injected with 3×106UCBMC via the lateral ventricle.Oligodendrocytes and MBP protein in the corpus striatum of the injured brain were observed by CNPase+ Cleaved Caspase-3+ and MBP+DAPI+ immunofluorescence doule staining,and their correlation was analyzed. Results After transplantation 24 h,3 d and 7 d,there were more CNPase+ Cleaved Caspase-3+cells in WMD group than the CON,WMD,WMD+UCBMC and UCBMC groups (P<0.01),and began to increase in 24 h,reached the peak in 3 d,there decreased in 7 d,CNPase+ Cleaved Caspase-3+ cells of WMD+UCBMC group was significantly less than the WMD group (P<0.01).14 days after transplation,MBP+DAPI+ cells of the WMD group were significantly less than those of the CON and UCBMC control groups (P<0.01),MBP+DAPI+ cells of the WMD+UCBMC group were significantly higher than those in the WMD group,but still less than those of the CON group (P<0.01). Conclusions UCBMC transplantation via the lateral ventricle could reduce the oligodendrocytes alleviate,thus repair the white matter in neonatal rats after WMD.
关键词
脐血单个核细胞 /
脑白质 /
髓鞘碱性蛋白 /
少突胶质细胞 /
低龄大鼠
Key words
umbilical cord blood monocytes /
white matter /
myelin basic protein /
oligodendrocytes /
low-age rats
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基金
国家自然科学基金项目(81000268)；山东省自然科学基金项目(ZR2010HQ037)；山东省自然科学基金项目(ZR2013HL0670)
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