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目的观察经不同途径移植脂肪来源干细胞(ADSC)后创伤性脑损伤大鼠记忆的变化,探讨干细胞移植的最佳途径。方法以酶化学法自大鼠脂肪组织中分离培养大鼠ADSCs,建立侧方液压打击创伤性脑损伤大鼠模型,将ADSCs分别经尾静脉注射(伤后1d、3d、7d,2×106/次)、海马背侧CA1区局部注射(伤后1d,2×106/次)移植入实验动物体内,对照组造模成功后,不予处理自然转归,各组模型均于移植后22天采用Morris水迷宫检测大鼠的空间记忆功能以及伤后7d、14d、28d应用RT-PCR检测脑源性神经生长因子(BDNF)m RNA的表达。结果经酶消化法分离得到的ADSCs经流式细胞检测以及成骨、成脂分化提示其具有间充质干细胞的生物学特性,经不同途径移植ADSCs后,定位航行测试结果表明细胞移植组逃避潜伏期均较对照组明显缩短(P<0.05);空间探索能力测试结果表明细胞移植组于目标象限的游泳时间较对照组明显延长(P<0.05),RT-PCR检测结果表明细胞移植组BDNF m RNA的表达较对照组明显增高,差别均具有统计学意义(P<0.05),而移植组之间在空间能力的改善以及14、28天时BDNF m RNA的表达的影响无统计学差异(P>0.05)。结论治疗大鼠创伤性脑损伤时,ADSCs经尾静脉(多次移植)与脑内损伤区移植在改善空间记忆障碍方面无显著差异。
Objective To observe the changes of memory in traumatic brain injury rats after transplanted with adipose-derived stem cells (ADSCs) by different ways, and to explore the best way of stem cell transplantation. Methods Rat ADSCs were isolated and cultured from rat adipose tissue by enzymatic chemistry method. A rat model of lateral hydraulic attack on traumatic brain injury was established. The ADSCs were injected through tail vein (1 d, 3 d, 7 d, 2 × 10 6 / Times). The CA1 area of the dorsal hippocampus was transplanted into the experimental animals by local injection (2 × 106 / time 1d after injury). After the model was established in the control group, the natural outcome was not treated. Morris water maze was used to detect the spatial memory function of rats and RT-PCR was used to detect the expression of brain derived nerve growth factor (BDNF) m RNA at 7d, 14d, 28d after injury. Results ADSCs isolated by enzyme digestion method showed the biological characteristics of mesenchymal stem cells by flow cytometry and osteogenic and adipogenic differentiation. After transplanted ADSCs by different routes, the positioning navigation test results showed that the cell migration group escape latency (P <0.05). The results of spatial exploration test showed that the swimming time in the target quadrant of the cell transplantation group was significantly longer than that in the control group (P <0.05). The RT-PCR results showed that the BDNF m RNA (P <0.05), while there was no significant difference in the improvement of spatial ability between the transplantation group and the expression of BDNF m RNA at 14 and 28 days (P> 0.05) ). Conclusion In the treatment of traumatic brain injury in rats, ADSCs transplanted through tail vein (multiple transplants) and intracerebral injury area have no significant difference in improving spatial memory impairment.