川芎嗪联合骨髓间充质干细胞移植抑制脑缺血大鼠神经细胞凋亡

发布时间：2018-10-13 12:40

【摘要】：目的:研究川芎嗪(tetramethylpyrazine,TMP)联合骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)对脑缺血大鼠神经细胞凋亡及Bcl-2、Bax表达的影响。方法:采用全骨髓贴壁法体外培养BMSCs,传代至第3代用于尾静脉移植。采用线栓法诱导大鼠右侧大脑中动脉阻塞模型,除假手术组外,大鼠随机分为模型组、BMSCs(1×10~9/L)组、川芎嗪(40 mg/kg)组和联合(川芎嗪+BMSCs)组,每组12只。缺血后第1、7和14天采用改良的神经损伤严重程度评分(modified neurological severity scoring,m NSS)进行神经功能评价。缺血后第14天,甲苯胺蓝染色检测脑梗死体积,HE染色观察脑组织病理学变化,采用原位末端标记(TUNEL)法观察神经细胞凋亡数,采用实时荧光定量PCR法和Western blot法检测Bax和Bcl-2的mRNA及蛋白表达。结果:与BMSCs组和川芎嗪组比较,联合组m NSS评分显著减少(P0.01),梗死体积显著减少(P0.01),缺血引起的脑缺血周边区病理性损伤明显减轻,TUNEL阳性细胞数显著减少(P0.01),Bcl-2的mRNA及蛋白表达显著增加,Bax的mRNA及蛋白表达显著降低(P0.01)。结论:川芎嗪联合BMSCs移植能显著促进脑缺血后大鼠的功能恢复,减少梗死体积,减轻脑组织缺血性损伤,抑制神经细胞凋亡,机制可能与调控Bcl-2和Bax的表达有关。
[Abstract]:Aim: to study the effects of ligustrazine (tetramethylpyrazine,TMP) combined with bone marrow mesenchymal stem cell (bone marrow mesenchymal stem cells,BMSCs) on neuronal apoptosis and Bcl-2,Bax expression in rats with cerebral ischemia. Methods: the whole bone marrow adherent method was used to culture BMSCs, for caudal vein transplantation. The right middle cerebral artery occlusion model was induced by thread occlusion in rats. Except sham-operation group, rats were randomly divided into three groups: model group (, BMSCs (1 脳 10 9 / L), ligustrazine (40 mg/kg) group and ligustrazine BMSCs group (12 rats in each group). The neurological function was evaluated on the 7th and 14th day after ischemia by using the modified nerve injury severity score (modified neurological severity scoring,m NSS). On the 14th day after ischemia, toluidine blue staining was used to detect the volume of cerebral infarction, HE staining was used to observe the pathological changes of brain tissue, and the number of neuronal apoptosis was observed by in situ end labeling (TUNEL) method. The expression of mRNA and protein in Bax and Bcl-2 were detected by real-time fluorescence quantitative PCR and Western blot. Results: compared with BMSCs group and ligustrazine group, In the combined group, the score of m NSS was significantly decreased (P0.01), the infarct volume was significantly decreased (P0.01), the pathological injury of ischemic peripheral area was alleviated significantly, the number of TUNEL positive cells was significantly decreased (P0.01), the expression of mRNA and protein in Bcl-2 was significantly increased, and the expression of mRNA and protein in Bax was significantly decreased (P0.01). Conclusion: ligustrazine combined with BMSCs transplantation can significantly promote functional recovery, reduce infarct volume, reduce ischemic injury of brain tissue and inhibit neuronal apoptosis in rats after cerebral ischemia. The mechanism may be related to the regulation of the expression of Bcl-2 and Bax.
【作者单位】：浙江中医药大学药学院;浙江中医药大学基础医学院;
【基金】：国家自然科学基金资助项目(No.81274113) 浙江省公益技术应用研究计划(No.2016C33185) 浙江省教育厅科研项目(No.Z201016310)
【分类号】：R743.3

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