In order to investigate the protective effects of the overexpression of bcl-xl gene on local cerebral infarction in the transgenic mice subject to permanent occlusion of middle cerebral artery, the models of bcl-xl tr...In order to investigate the protective effects of the overexpression of bcl-xl gene on local cerebral infarction in the transgenic mice subject to permanent occlusion of middle cerebral artery, the models of bcl-xl transgenic mice were established and subjected to cerebral infarction by intraluminal occlusion of the middle cerebral artery. The infarct volume and the neurological scores were observed and comparison between the wild type mice and the transgenic mice was made. It was found that the infarct volume and the neurological scores in the transgenic mice were significantly decreased as compared with those in the wild type mice. It was suggested that the overexpression of bcl-xl gene in transgenic mice could reduce the infarct volume and improve the neurological function of the mice.展开更多
The efficiency and safe range of LipofectamineTM2000 (LF2000)/bcl-xl applied in human keratocytes, the optimal ratio of LF2000/bcl-xl and the bcl-xl gene expression in human keratocytes were investiaged. By using tryp...The efficiency and safe range of LipofectamineTM2000 (LF2000)/bcl-xl applied in human keratocytes, the optimal ratio of LF2000/bcl-xl and the bcl-xl gene expression in human keratocytes were investiaged. By using trypan-blue staining, the effects of LF2000 and bcl-xl on the survival rate of the cultured human keratocytes were measured respectively. By using semi-quantitative RT-PCR, the efficiency and the expression of LF2000-mediated bcl-xl transfection into keratocytes were examined. The results showed that the survival rate of human keratocytes had no signficant change in the presence of LF2000 (20 μg/ml) or bcl-xl (10 μg/ml) for 24 h. LF2000 could effectively mediate the transfection of exogenous gene bcl-x1 into human keratocytes. The best transfection efficiency could be obtained when the ratio of bcl-xl/LF2000 was 1:8. One day after transfection, the positive cells for bcl-x1 could be detectable, and the positive rate reached the peak on the post-transfection day 3 (48.3 %), then gradually decreased. Fifteen days after transfection, there were few positive cells. It was suggested that LF2000 could effectively transfer the exogenous gene bcl-xl into human keratocytes without obvious toxicity during a concentration range. LF2000/bcl-xl may be likely to play an important role in gene therapy of human keratocytes.展开更多
目的 观察 bcl- x L 基因可否介导白血病细胞的多药耐药 ,并探讨其机制 .方法 采用脂质体法将 bcl- x L c DNA转导入 HL- 6 0细胞 ;免疫印迹法检测 Bcl- x L 及抗凋亡蛋白Bax的表达 ;MTT法测定足叶乙甙、柔红霉素、阿霉素对转染细胞...目的 观察 bcl- x L 基因可否介导白血病细胞的多药耐药 ,并探讨其机制 .方法 采用脂质体法将 bcl- x L c DNA转导入 HL- 6 0细胞 ;免疫印迹法检测 Bcl- x L 及抗凋亡蛋白Bax的表达 ;MTT法测定足叶乙甙、柔红霉素、阿霉素对转染细胞的细胞毒性 ;流式细胞仪定量检测凋亡细胞及细胞内药物浓度 .结果 Bcl- x L高表达可抑制足叶乙甙诱发的凋亡 ,抑制率为 40 .5 % ;降低上述化疗药物的细胞毒作用 ,耐药指数分别为 3.2 ,3.2 ,1 .6 ;但细胞内柔红霉素药物浓度无改变 .结论 Bcl- x L可能作为一种新的耐药途径 ,通过抑制化疗药物诱发的细胞凋亡参与多药耐药的形成 .展开更多
基金a grant from National Natu-ral Sciences Foundation of China (No. 30500167)
文摘In order to investigate the protective effects of the overexpression of bcl-xl gene on local cerebral infarction in the transgenic mice subject to permanent occlusion of middle cerebral artery, the models of bcl-xl transgenic mice were established and subjected to cerebral infarction by intraluminal occlusion of the middle cerebral artery. The infarct volume and the neurological scores were observed and comparison between the wild type mice and the transgenic mice was made. It was found that the infarct volume and the neurological scores in the transgenic mice were significantly decreased as compared with those in the wild type mice. It was suggested that the overexpression of bcl-xl gene in transgenic mice could reduce the infarct volume and improve the neurological function of the mice.
文摘The efficiency and safe range of LipofectamineTM2000 (LF2000)/bcl-xl applied in human keratocytes, the optimal ratio of LF2000/bcl-xl and the bcl-xl gene expression in human keratocytes were investiaged. By using trypan-blue staining, the effects of LF2000 and bcl-xl on the survival rate of the cultured human keratocytes were measured respectively. By using semi-quantitative RT-PCR, the efficiency and the expression of LF2000-mediated bcl-xl transfection into keratocytes were examined. The results showed that the survival rate of human keratocytes had no signficant change in the presence of LF2000 (20 μg/ml) or bcl-xl (10 μg/ml) for 24 h. LF2000 could effectively mediate the transfection of exogenous gene bcl-x1 into human keratocytes. The best transfection efficiency could be obtained when the ratio of bcl-xl/LF2000 was 1:8. One day after transfection, the positive cells for bcl-x1 could be detectable, and the positive rate reached the peak on the post-transfection day 3 (48.3 %), then gradually decreased. Fifteen days after transfection, there were few positive cells. It was suggested that LF2000 could effectively transfer the exogenous gene bcl-xl into human keratocytes without obvious toxicity during a concentration range. LF2000/bcl-xl may be likely to play an important role in gene therapy of human keratocytes.
文摘目的 观察 bcl- x L 基因可否介导白血病细胞的多药耐药 ,并探讨其机制 .方法 采用脂质体法将 bcl- x L c DNA转导入 HL- 6 0细胞 ;免疫印迹法检测 Bcl- x L 及抗凋亡蛋白Bax的表达 ;MTT法测定足叶乙甙、柔红霉素、阿霉素对转染细胞的细胞毒性 ;流式细胞仪定量检测凋亡细胞及细胞内药物浓度 .结果 Bcl- x L高表达可抑制足叶乙甙诱发的凋亡 ,抑制率为 40 .5 % ;降低上述化疗药物的细胞毒作用 ,耐药指数分别为 3.2 ,3.2 ,1 .6 ;但细胞内柔红霉素药物浓度无改变 .结论 Bcl- x L可能作为一种新的耐药途径 ,通过抑制化疗药物诱发的细胞凋亡参与多药耐药的形成 .