携带ApoJ基因的骨髓间充质干细胞治疗大鼠脑出血模型的研究

发布时间：2018-06-16 03:11

  本文选题：载脂蛋白J(ApoJ) + 脑出血　； 参考：《西南医科大学》2017年硕士论文

【摘要】：目的:探讨携带ApoJ基因的骨髓间充质干细胞对大鼠实验性脑出血的治疗作用。方法:采取脂质体介导用重组质粒pEGFP-N1-apoJ转染体外培养的骨髓间充质干细胞(BMSCs),分别于转染后24,48,72,96h在荧光显微镜下观察相同体积脂质体介导不同重组质粒量的转染效果,筛选出转染率最高的转染组。将102只雄性SD大鼠随机分为转染组、BMSCs组和NaCl组。每组再分为24h,48h,72h,96h四个亚组。利用脑立体定位仪建立大鼠脑出血模型,三组分别于造模24h后颅内注射等体积的转染细胞悬液、BMSCs悬液和生理盐水,分别于处理后24h,48h,72h,96h处死各对应亚组大鼠。进行各时间点改良神经功能评分(mNSS);用干湿质量法测量脑组织含水量;用蛋白免疫印迹法检测载脂蛋白-J在各时间点的表达。结果:1.等体积脂质体条件下,四个细胞转染组(重组质粒量分别为0.8、1.2、1.6、2.0)在72h时转染率高于同组在24h,48h,96h的转染率(P均0.05);且与另外三个细胞转染组(重组质粒量分别为0.8、1.2、2.0)比较,重组质粒量为1.6的细胞转染组在72h时荧光表达最强,转染效率高于同时间点的其他三组(P均0.05);2.mNSS结果显示,转染组在实验干预后1,3,5,7d得分显著低于同时间点的BMSCs组与NaCl组(P均0.05);3.脑组织含水量测定结果显示,转染组在1,3,5,7d脑组织含水量低于同时间点的BMSCs组与NaCl组(P均0.05);4.Western blot结果显示,与BMSCs组与NaCl组比较,转染组脑组织内ApoJ表达水平均高于同时间点的两组(P均0.05)。结论:携带ApoJ基因的骨髓间充质干细胞能够在ICH大鼠脑组织内表达载脂蛋白J,外源性ApoJ对脑出血具有治疗作用,可以促进脑出血大鼠的神经功能的恢复,减轻脑水肿,改善脑出血大鼠预后。
[Abstract]:Aim: to investigate the therapeutic effect of bone marrow mesenchymal stem cells carrying ApoJ gene on experimental intracerebral hemorrhage in rats. Methods: the recombinant plasmid pEGFP-N1-apoJ was transfected into bone marrow mesenchymal stem cells (BMSCs) by liposome-mediated transfection in vitro. The transfection effects of different amount of recombinant plasmids mediated by the same volume of liposome were observed under fluorescence microscope at 24: 48 ~ 72 ~ 96 h after transfection. The transfection group with the highest transfection efficiency was selected. 102 male SD rats were randomly divided into BMSCs group and NaCl group. Each group was subdivided into four subgroups: 24 h, 48 h, 72 h and 96 h. The intracerebral hemorrhage model of rats was established by brain stereotactic locator. BMSCs suspension and normal saline were injected into the brain of the three groups 24 hours after the establishment of the model. The rats of the corresponding subgroups were killed at 24 h, 48 h, 72 h and 96 h, respectively. The modified neurologic function scores were evaluated at each time point, the water content of brain tissue was measured by wet and dry mass method, and the expression of apolipoprotein J at each time point was detected by Western blot. The result is 1: 1. In the same volume liposome condition, the transfection efficiency of the four cell transfection groups (the amount of recombinant plasmids was 0.81.21.62.0) at 72 h was higher than that in the same group at 24 h, 48 h or 96 h, and was compared with the other three cell transfection groups (the amount of recombinant plasmids was 0.88 1.22 2.0). The transfection group with 1.6 recombinant plasmid had the strongest fluorescence expression at 72 h, and the transfection efficiency was higher than that of the other three groups at the same time point (P 0.05) and 2.mNSS. The results showed that the score of transfection group was significantly lower than that of BMSCs group and NaCl group on the 1st and 5th day after experimental intervention. The water content of brain tissue in transfection group was lower than that in BMSCs group and NaCl group at 1 ~ 3 ~ 5 ~ 5 ~ 5 ~ 7 days after transfection. 4. The results showed that the water content of brain tissue in transfection group was lower than that in BMSCs group and NaCl group at the same time point. 4. The results showed that compared with BMSCs group and NaCl group. The expression of ApoJ in brain tissue of the transfected group was higher than that of the two groups at the same time point (P < 0.05). Conclusion: bone marrow mesenchymal stem cells carrying ApoJ gene can express apolipoprotein J in the brain of ICH rats. Exogenous ApoJ can promote the recovery of neural function and alleviate cerebral edema in ICH rats. To improve the prognosis of rats with intracerebral hemorrhage.
【学位授予单位】：西南医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R743.34

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