BMSCs移植联合电针刺激治疗对脑出血大鼠脑内骨形态发生蛋白、胶质细胞和血管再生的影响

发布时间：2018-06-06 16:35

  本文选题：骨髓间充质干细胞 + 电针　； 参考：《泸州医学院》2014年硕士论文

【摘要】：目的通过骨髓间充质干细胞(bone marrow mesenchymal stem cells, BMSCs)移植联合电针(electric acupuncture, Ea)刺激治疗脑出血大鼠,观察出血灶及周边骨形态发生蛋白-2(bone morphogenetic protein-2, BMP2)、神经胶质抗原-2(neuroglia antigen-2, NG2)、半乳糖脑苷脂(galactocerebroside, GALC)、CD68蛋白及层粘连蛋白(laminin, LN)的表达变化,探讨联合治疗对骨形态发生蛋白、脑内胶质细胞和血管再生影响。方法100只雄性SD大鼠建立脑出血模型,随机分BMSCs移植联合电针治疗组(BMSCs+Ea组)、MSCs移植组(BMSCs组)、电针治疗组(Ea组)和生理盐水组(NS组),每组25只。术后第三天分别给予BMSCs移植联合电针刺激、BMSCs移植、电针刺激和注射生理盐水治疗；又根据处死时间的不同,每组大鼠又分成1d、3d、5d、7d、14d5个亚组,每亚组5只。取脑制成冰冻切片,免疫组化检测BMP2、NG2、GALC、CD68、LN蛋白的表达。结果1.出血灶周边脑区BMP2阳性细胞计数并分析：各组BMP2阳性细胞数先呈上升趋势而后下降。BMSCs+Ea组比NS组在1d-14d时增多,BMSCs组比NS组在3d-5d升高(P0.05)。Ea组和NS组相比较无显著差异。BMSCs+Ea组比Ea组在1d、5d和7d时增多,BMSCs组比Ea组在1d和5d时上升(P0.05)。BMSCs+Ea组与BMSCs组相比在1d和14d时上调(P0.05)。2.出血灶周边脑区NG2阳性细胞计数并分析结果显示：BMSCs+Ea组比NS组在1d-7d时增多,BMSCs组比NS组在5d和14d升高(P0.05)。Ea组与NS组相比差异无统计学意义。BMSCs+Ea组比Ea组在1d、3d、5d和14d时增多,BMSCs组比Ea组5d和14d时上升(P0.05)。BMSCs+Ea组与BMSCs组相比在1d-5d时上调(P0.05)。3.出血灶周边脑区GALC阳性细胞计数后分析,BMSCs+Ea组比NS组在1d-7d时增多,BMSCs组比NS组在1d-7d升高,Ea组比NS组在7d时上升(P0.05)。BMSCs+Ea组比Ea组在1d-5d时增多,BMSCs组比Ea组在5d时上升(P0.05)。BMSCs+Ea组与BMSCs组相比在1d-3d时上调(P0.05)。4.出血灶周边脑区CD68蛋白光密度值变化：BMSCs+Ea组、BMSCs组和Ea组CD68蛋白光密度值先缓慢上升而后逐渐下降,7d时达到峰值。NS组在1d-14d均呈上升趋势。BMSCs+Ea组比NS组在1d-14d时增大, BMSCs组比NS组在1d-5d升高,在14d时下降,Ea组比NS组在14d时减小(P0.05)。BMSCs+Ea组比Ea组在1d-7d时增大(P0.05)。BMSCs组比Ea组相比较差异无统计学意义。BMSCs+Ea组与BMSCs组相比在5d-7d时上调(P0.05)。5.对出血灶周边脑区血管计数并分析：BMSCs+Ea组血管数比NS组在5d-14d时增多,BMSCs组比NS组在3d-7d升高,Ea组比NS组在3d时上升(P0.05)。BMSCs+Ea组比Ea组在5d-14d时增多,BMSCs组比Ea组5d-14d时上升(P0.05)。BMSCs+Ea组与BMSCs组相比较在7d时上调(P0.05)。结论1.BMSCs移植联合电针治疗较单纯的BMSCs移植和电针刺激治疗更能促进脑出血大鼠BMP2的表达,这有助于脑内NSCs和BMSCs向神经细胞方向转化及脑组织结构重塑。2.BMSCs移植联合电针治疗早期可促进少突胶质前体细胞和少突胶质细胞的增殖,利于髓鞘形成；后期则抑制其增殖,可能与防止胶质瘢痕的形成有关。3.BMSCs移植可早期激活小胶质细胞,促进其快速增殖；联合电针治疗后早期上调其活化和增殖的能力,积极参与免疫反应。4.单纯BMSCs移植和电针刺激治疗可在一定时间内促进血管再生；BMSCs移植联合电针治疗更有利于血管的生成,有助于改善出血灶及周边区微环境。
[Abstract]:Objective to treat cerebral hemorrhage rats by transplantation of bone marrow mesenchymal stem cells (BMSCs) and electroacupuncture (electric acupuncture, Ea), and to observe the hemorrhagic foci and peripheral bone morphogenetic protein -2 (bone morphogenetic), glial antigen, and galactoside. The expression of galactocerebroside (GALC), CD68 protein and laminin (laminin, LN) and the effect of combined therapy on bone morphogenetic protein, brain glial cells and vascular regeneration. Methods 100 male SD rats were established with cerebral hemorrhage model, randomly divided into BMSCs graft combined electroacupuncture group (BMSCs+Ea group) and MSCs transplantation group (BMSCs group). Electroacupuncture treatment group (group Ea) and saline group (group NS), 25 in each group, were given BMSCs transplantation combined with electroacupuncture stimulation, BMSCs transplantation, electroacupuncture stimulation and injection of saline for third days after operation, and the rats in each group were divided into 1D, 3D, 5D, 7d, 14d5 subgroups and 5 in each subgroup according to the time of death. The brain was made into frozen section and immune group. Results 1. the expression of BMP2, NG2, GALC, CD68, LN protein. Results the number of BMP2 positive cells in the peripheral brain region of the hemorrhagic focus and analysis: the number of BMP2 positive cells in each group first increased and then decreased in the.BMSCs+Ea group than in the NS group at 1d-14d, and there was no significant difference in the BMSCs group compared with the NS group. The increase of 1D, 5D and 7d, the increase of BMSCs group than that of Ea in 1D and 5D (P0.05).BMSCs+Ea group compared with BMSCs group in 1D and 14d. The.BMSCs+Ea group was more than the Ea group at 1D, 3D, 5D and 14d, and the BMSCs group was higher than the Ea group 5D and 14d. The increase of (P0.05).BMSCs+Ea group was more than that of the Ea group at 1d-5d, and the BMSCs group was higher than that of the Ea group at 5D (P0.05) in the.BMSCs+Ea group compared with the BMSCs group. Group S increased in the 1d-14d group than in the NS group at 1d-14d, and the BMSCs group was higher than the NS group in 1d-5d, and decreased at 14d, and the Ea group was smaller than the NS group in 14d. The number of blood vessels in the peripheral brain region of the hemorrhagic foci was increased by.5.. The number of blood vessels in the group BMSCs+Ea was higher than that in the NS group at 5d-14d, and the BMSCs group was higher than the NS group in 3d-7d, and the Ea group was higher than the NS group at 3D. 1.BMSCs transplantation combined with electroacupuncture can promote the expression of BMP2 in rats with intracerebral hemorrhage more than simple BMSCs transplantation and electroacupuncture stimulation, which helps to transform NSCs and BMSCs into neurons in the brain and to reshape the structure of brain tissue and remodeling.2.BMSCs transplantation combined with electroacupuncture at the early stage to promote the increase of oligodendrocytes and oligodendrocytes. Colonization is beneficial to the formation of myelin sheath; later, it inhibits its proliferation. It may be related to the prevention of the formation of glial scar..3.BMSCs transplantation can activate microglia early and promote its rapid proliferation. The ability to increase its activation and proliferation early after electroacupuncture treatment, and actively participate in the immune response.4. simple BMSCs transplantation and electroacupuncture stimulation treatment may be certain. BMSCs transplantation combined with electro acupuncture treatment is more conducive to angiogenesis and helps to improve the microenvironment of bleeding foci and surrounding areas.
【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R743.34

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