CAJ | 학술논문

目的探讨骨髓单个核细胞(BMMNCs)移植治疗对脑出血大鼠胶质源性神经营养因子(GDNF)的表达及其脑内神经元样细胞分化情况的影响. 方法纹状体区立体定向注射胶原酶建立脑出血模型,筛选成功模型后随机分为脑出血组和BMMNCs组,后者于造模后24 h经尾静脉注入1mL5-溴脱氧尿嘧啶核苷(Brdu)标记的含3×107个BMMNCs细胞悬液,再于移植后1d、7d、14d、28d采用改良的大鼠神经功能评分(mNSS)观察大鼠神经行为学表现;于移植后的1d、7d、14d采用Western blotting方法检测脑内GDNF的表达情况;28 d后采用免疫组织荧光双标法示踪BMMNCs在脑内分化情况. 结果移植后7d、14d、28 d,BMMNCs组mNSS评分均明显低于脑出血组,脑出血组各时间点GDNF表达量明显低于BMMNCs组,差异均有统计学意义(P＜0.05);术后28 d,BMMNCs组可见Brdu和BMMNCs双标阳性细胞. 结论 BMMNCs移植能显著改善大鼠神经行为学功能,明显增加GDNF的分泌量,并能在脑出血大鼠体内分化为神经元样细胞.
목적 탐토골수단개핵세포(BMMNCs)이식치료대뇌출혈대서효질원성신경영양인자(GDNF)적표체급기뇌내신경원양세포분화정황적영향. 방법 문상체구입체정향주사효원매건립뇌출혈모형,사선성공모형후수궤분위뇌출혈조화BMMNCs조,후자우조모후24 h경미정맥주입1mL5-추탈양뇨밀정핵감(Brdu)표기적함3×107개BMMNCs세포현액,재우이식후1d、7d、14d、28d채용개량적대서신경공능평분(mNSS)관찰대서신경행위학표현;우이식후적1d、7d、14d채용Western blotting방법검측뇌내GDNF적표체정황;28 d후채용면역조직형광쌍표법시종BMMNCs재뇌내분화정황. 결과 이식후7d、14d、28 d,BMMNCs조mNSS평분균명현저우뇌출혈조,뇌출혈조각시간점GDNF표체량명현저우BMMNCs조,차이균유통계학의의(P＜0.05);술후28 d,BMMNCs조가견Brdu화BMMNCs쌍표양성세포. 결론 BMMNCs이식능현저개선대서신경행위학공능,명현증가GDNF적분비량,병능재뇌출혈대서체내분화위신경원양세포.
Objective To investigate the effect of transplantation of bone marrow mononuclear cells (BMMNCs) on neuron-like cell differentiation and glial cell-derived neurotrophie factor (GDNF) expression in rats with intracerebral hemorrhage (ICH).Methods ICH was induced by stereotactic injection of type Ⅳ collagenase into the striatum of rats,and the successful models were randomly divided into model group (ICH group) and BMMNCs transplantation group (BMMNCs group); BMMNCs group was injected with 1 mL cell suspension containing 3 ×107 cells marked with 5-Bromo-2-deoxyuridine (Brdu).The neurological functions of the rats were observed by modified neurological severity scale (mNSS) test 1,7,14 and 28 days after the injection; the GDNF expression in cerebral tissue was observed by Western blotting 1,7,14 days after the injection; double immunofluorescent labeling was utilized to trace the differentiation of BMMNCs in the rat brains 28 days after the injection.Results As compared with those in the ICH group,the mNSS scores in the BMMNCs group were significantly lower (P＜0.05) and the GDNF expression was significantly increased at all time points (P＜0.05); Brdu and neuron specific nuclear protein (NeuN) double-labeled positive cells were found in the hemorrhage focus in the BMMNCs group 28 d after injection.Conclusion BMMNCs transplantation can improve the neural behavioral function,increase the GDNF expression,and induce BMMNCs differentiating into neuron-like cells.