目的建立real time逆转录聚合酶链反应(RT-PCR)检测BDNF mRNA基因表达的方法.方法提取脑缺血组织的总RNA,进行RT-PCR扩增BDNF mRNA特异性片段,扩增产物重组到质粒上并测序,建立real time RT-PCR检测BDNF mRNA表达水平方法.结果重组的质...目的建立real time逆转录聚合酶链反应(RT-PCR)检测BDNF mRNA基因表达的方法.方法提取脑缺血组织的总RNA,进行RT-PCR扩增BDNF mRNA特异性片段,扩增产物重组到质粒上并测序,建立real time RT-PCR检测BDNF mRNA表达水平方法.结果重组的质粒经酶切和测序,目的片段已插入到载体内,得到real time RT-PCR动力学曲线.结论成功建立real time RT-PCR检测BDNF mRNA基因表达的方法.展开更多
目的探讨MicroRNA-101(miRNA-101)对上皮性卵巢癌耐药细胞SKOV3/DDP顺铂敏感性的影响及相关作用机制。方法本研究通过细胞转染技术上调SKOV3/DDP细胞内miRNA-101的表达,采用RT-PCR检测miRNA-101及BDNF m RNA的表达情况;CCK-8检测SKOV3/...目的探讨MicroRNA-101(miRNA-101)对上皮性卵巢癌耐药细胞SKOV3/DDP顺铂敏感性的影响及相关作用机制。方法本研究通过细胞转染技术上调SKOV3/DDP细胞内miRNA-101的表达,采用RT-PCR检测miRNA-101及BDNF m RNA的表达情况;CCK-8检测SKOV3/DDP细胞顺铂敏感性及增殖力;Annexin V-FITC/PI流式细胞实验检测细胞凋亡率。结果与阴性对照组比较,miRNA-101转染组细胞miRNA-101表达量及细胞凋亡率增加[(1.000±0.022)vs(5.380±0.246),P<0.05],[(11.020±0.685)%vs(26.158±1.278)%,P<0.05];BDNF m RNA表达量降低[(1.000±0.042)vs(0.389±0.055),P<0.05];顺铂IC50及细胞增殖力均下降[(57.276±1.717)vs(33.176±2.465),P<0.05],[(1.776±0.030)vs(1.642±0.252),P<0.05]。结论 miRNA-101可有效增加SKOV3/DDP细胞对顺铂的药物敏感性,促进细胞凋亡。展开更多
The concept of a major inhibitory role of serotonin in aggressive behavior is widely accepted by investigators. There was ample evidence that a pharmacologically-induced increase in the serotonin activity attenuates a...The concept of a major inhibitory role of serotonin in aggressive behavior is widely accepted by investigators. There was ample evidence that a pharmacologically-induced increase in the serotonin activity attenuates agonistic behavior in animals, and that the manipulations inhibiting the brain serotonergic system can elicit aggressiveness in male mice and rats. Repeated experience of aggression in daily agonistic interactions has been shown to reduce serotonin activity in brain of victorious male mice. The study aimed to analyze expression of the serotonergic genes -Tph2, Sert, Maoa and Htr1a, as well as the Bdnf and Creb genes in the midbrain raphe nuclei of male mice with positive fighting experience in daily encounters and male mice with the same track record of aggression followed by two-week no-fight period. It has been shown that mRNA levels of the serotonergic and Creb genes are reduced in the winners in comparison with male mice without consecutive positive fighting experience of aggression. After the fighting deprivation the Tph2, Sert, Bdnf and Creb genes recover their expression while mRNA levels of the Maoa and Htr1a genes proceed at a significantly higher level as compared with the respective controls. Downregulation of serotonergic genes is indicative of the inhibition of serotonergic activity under repeated experience of aggression. Nevertheless recovering of Tph2 and Sert gene expression and overexpression of the Maoa and Htr1a genes after no-fight period suggest that changes in brain serotonergic activity are not main cause of the behavioral pathology developing in male mice in this experimental context.展开更多
文摘目的建立real time逆转录聚合酶链反应(RT-PCR)检测BDNF mRNA基因表达的方法.方法提取脑缺血组织的总RNA,进行RT-PCR扩增BDNF mRNA特异性片段,扩增产物重组到质粒上并测序,建立real time RT-PCR检测BDNF mRNA表达水平方法.结果重组的质粒经酶切和测序,目的片段已插入到载体内,得到real time RT-PCR动力学曲线.结论成功建立real time RT-PCR检测BDNF mRNA基因表达的方法.
文摘目的探讨MicroRNA-101(miRNA-101)对上皮性卵巢癌耐药细胞SKOV3/DDP顺铂敏感性的影响及相关作用机制。方法本研究通过细胞转染技术上调SKOV3/DDP细胞内miRNA-101的表达,采用RT-PCR检测miRNA-101及BDNF m RNA的表达情况;CCK-8检测SKOV3/DDP细胞顺铂敏感性及增殖力;Annexin V-FITC/PI流式细胞实验检测细胞凋亡率。结果与阴性对照组比较,miRNA-101转染组细胞miRNA-101表达量及细胞凋亡率增加[(1.000±0.022)vs(5.380±0.246),P<0.05],[(11.020±0.685)%vs(26.158±1.278)%,P<0.05];BDNF m RNA表达量降低[(1.000±0.042)vs(0.389±0.055),P<0.05];顺铂IC50及细胞增殖力均下降[(57.276±1.717)vs(33.176±2.465),P<0.05],[(1.776±0.030)vs(1.642±0.252),P<0.05]。结论 miRNA-101可有效增加SKOV3/DDP细胞对顺铂的药物敏感性,促进细胞凋亡。
文摘The concept of a major inhibitory role of serotonin in aggressive behavior is widely accepted by investigators. There was ample evidence that a pharmacologically-induced increase in the serotonin activity attenuates agonistic behavior in animals, and that the manipulations inhibiting the brain serotonergic system can elicit aggressiveness in male mice and rats. Repeated experience of aggression in daily agonistic interactions has been shown to reduce serotonin activity in brain of victorious male mice. The study aimed to analyze expression of the serotonergic genes -Tph2, Sert, Maoa and Htr1a, as well as the Bdnf and Creb genes in the midbrain raphe nuclei of male mice with positive fighting experience in daily encounters and male mice with the same track record of aggression followed by two-week no-fight period. It has been shown that mRNA levels of the serotonergic and Creb genes are reduced in the winners in comparison with male mice without consecutive positive fighting experience of aggression. After the fighting deprivation the Tph2, Sert, Bdnf and Creb genes recover their expression while mRNA levels of the Maoa and Htr1a genes proceed at a significantly higher level as compared with the respective controls. Downregulation of serotonergic genes is indicative of the inhibition of serotonergic activity under repeated experience of aggression. Nevertheless recovering of Tph2 and Sert gene expression and overexpression of the Maoa and Htr1a genes after no-fight period suggest that changes in brain serotonergic activity are not main cause of the behavioral pathology developing in male mice in this experimental context.
