骨髓间充质干细胞对制动大鼠骨骼肌细胞Bcl-2、Bax蛋白表达的影响

发布时间：2019-04-07 20:04

【摘要】：目的观察早期注射骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)对制动大鼠骨骼肌细胞Bcl-2、Bax蛋白表达的影响方法取幼龄大鼠(80-100g)胫骨和股骨,分离培养大鼠BMSCs。用可塑性石膏制备右下肢管型固定肌萎缩模型,根据随机数字分组法将24只大鼠分成假手术组,干细胞组,PBS组,干细胞组大鼠下肢制动48h后比目鱼肌局部多点注射1×106个BMSCs;PBS组大鼠下肢制动48h后注射等量磷酸缓冲液(phosphate-buffered saline,PBS);假手术组大鼠下肢包裹相同重量石膏且不限制关节活动,48h后注射等量PBS,14天后各组大鼠拆除石膏取标本分析。结果培养中的大鼠骨髓间充质干细胞形态以梭形细胞为主,呈放射状排列的细胞集落,细胞生长旺盛,可连续稳定传代10代以上。实验中第4代骨髓间充质干细胞表面标志CD44,CD90均呈阳性表达,阳性率分别为95.84%,96.00%,而CD34,CD45阳性率极低,其阳性表达率分别为3.81%,4.25%。Western blot结果:与假手术组大鼠比目鱼肌中Bax和Bcl-2蛋白表达量[(0.14±0.11);(0.81±0.06)]比较,干细胞组和PBS组Bax蛋白表达量[(0.41±0.08);(0.63±0.10)]明显升高,而两组Bcl-2蛋白表达量[(0.47±0.14);(0.22±0.13)]明显降低;干细胞组中Bax蛋白的表达较PBS组降低明显,而Bcl-2蛋白的表达量显著增高,差异有明显统计学意义意义(P0.05)。结论制动大鼠骨骼肌早期注射BMSCs,可引起抗凋亡蛋白Bcl-2上调、促凋亡蛋白Bax表达的下调。
[Abstract]:Objective to observe the effect of early injection of bone marrow mesenchymal stem cells (bone mesenchymal stem cells,BMSCs) on the expression of Bcl-2,Bax protein in skeletal muscle cells of rats. Methods BMSCs. was isolated from tibia and femur of young rats (80 g) and cultured in vitro. 24 rats were divided into three groups: sham operation group, stem cell group, PBS group, stem cell group and stem cell group. After 48 hours of braking, 1 脳 10 6 BMSCs; were injected into the flounder muscle. Rats in PBS group were injected with equal phosphate buffer (phosphate-buffered saline,PBS) 48 hours after braking. The lower extremities of the sham-operated rats were covered with the same weight gypsum and the joint activity was not restricted. 48 hours after the injection of the same amount of PBS,14, the samples of the rats in each group were removed and analyzed. Results the cultured rat bone marrow mesenchymal stem cells (BMSCs) were mainly spindle-shaped cells with radially arranged colonies. The cells grew vigorously and could be continuously and stably passaged for more than 10 generations. The positive rate of CD44,CD90 was 95.84%, 96.0% respectively, while the positive rate of CD34,CD45 was very low, the positive rate was 3.81%, the positive rate of BMSCs was 95.84% and 96.0%, respectively, and the positive rate of BMSCs was 3.81%. 4.25%.Western blot results: compared with sham-operated group, the expression of Bax and Bcl-2 protein in soleus muscle [(0.14 卤0.11)]; (0.81 卤0.06), the expression of Bax protein in stem cell group and PBS group [(0.41 卤0.08); (0.63 卤0.10)] was significantly higher than that in control group [(0.47 卤0.14)], while the expression of Bcl-2 protein in both groups was significantly higher than that in control group [(0.47 卤0.14)]. The expression of Bax protein in stem cell group was significantly lower than that in PBS group, while the expression of Bcl-2 protein in stem cell group was significantly higher than that in PBS group (P0.05). Conclusion early injection of BMSCs, into skeletal muscle of rats can up-regulate the expression of anti-apoptotic protein Bcl-2 and down-regulate the expression of pro-apoptotic protein Bax.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R49

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