Transfection of STAT3 overexpression plasmid mediated through recombinant lentivirus promotes differentiation of bone marrow mesenchymal stem cells into neural cells in fetal rats with spina bifida aperta.
Transfection of STAT3 overexpression plasmid mediated through recombinant lentivirus promotes differentiation of bone marrow mesenchymal stem cells into neural cells in fetal rats with spina bifida aperta.
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重组慢病毒介导STAT3过表达质粒转染促进开放性脊柱裂胎鼠骨髓间充质干细胞向神经细胞分化
DOI：
10.18632/aging.203524
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发表时间：
2021-09-14
期刊：
影响因子：
--
通讯作者：
Ren M
中科院分区：
文献类型：
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作者：
Jiang M;Feng J;Fu R;Pan Y;Liu X;Dai J;Jiang C;Hao Y;Ren M
我们研究了信号转导和转录激活子-3(STAT3)对大鼠胚胎脊髓组织移植的影响。特别是，我们希望确定转导STAT3过表达质粒是否能增加脊髓移植的存活率，从而提高治疗效果。采用维甲酸建立大鼠脊柱裂模型，显微手术注射骨髓间充质干细胞(BMSCs)治疗。动物分为空白对照组、阴性对照组和实验组。用细胞计数试剂盒(CCK-8)测定BMSCs活性的光密度值。用实时荧光定量聚合酶链式反应和免疫印迹法检测STAT3、磷酸化STAT3、神经标志物和细胞凋亡相关因子的表达。实验组在CCK-8移植后8h的OD值最高。实验组pSTAT3、胶质纤维酸性蛋白、神经元特异性烯醇化酶、神经细丝和巢蛋白的表达明显高于空白对照组和阴性对照组(P<；0.05)。而实验组中STAT3的表达显著低于对照组(P<；0.05)。实验组caspase-8和bcl2的相对表达明显低于空白对照组和阴性对照组(P<；0.05)。将慢病毒介导的STAT3高表达载体与骨髓间充质干细胞共转染可提高转化为神经细胞的效率，为先天性脊柱裂的治疗提供新的种子细胞。
We investigated the influence of signal transducer and activator of transcription-3 (STAT3) on the spinal cord tissue grafts of rat fetuses with spina bifida aperta. In particular, we hoped to identify whether transfection of the STAT3 overexpression plasmid increases the survival of spinal cord transplantation in order to improve therapeutic efficacy. The fetal rat model of spina bifida aperta was established using retinoic acid and treated with a microsurgical injection of bone marrow mesenchymal stem cells (BMSCs). The animals were divided into either the blank control group, negative control group or the experimental group. The optical density (OD) value of BMSCs viability was determined using the Cell Counting Kit-8 (CCK-8). The expression of STAT3, phosphorylated STAT3 (pSTAT3), neural markers and apoptosis-related factors were evaluated using real-time PCR and Western blot. The OD value in the experimental group was highest at eight hours after transplantation using CCK-8. The expression of pSTAT3, glial fibrillary acidic protein, neuron-specific enolase, neurofilament and nestin in the experimental group was significantly higher compared to the blank control group and negative control group (P<0.05). However, STAT3 expression in the experimental group was statistically significantly decreased (P<0.05). The relative expression of caspase-8 and bcl-2 in the experimental group were significantly lower compared to the blank control group and negative control group (P<0.05). Transfection of the recombinant lentivirus-mediated STAT3 overexpression plasmid with BMSCs can help improve the efficiency of transforming into neural cells and provide new seed cells for the treatment of congenital spina bifida aperta.