鸟苷酸交换因子DOCK1对心肌细胞存活的影响

发布时间：2018-03-21 04:23

本文选题：鸟苷酸交换因子　切入点：DOCK1　出处：《重庆医科大学》2012年硕士论文　论文类型：学位论文

【摘要】：目的：探讨鸟苷酸交换因子DOCK1对心肌细胞存活的影响。方法：用重组真核质粒pCXN2-Flag-hDOCK1（人DOCK1过表达质粒），pCXN2-Flag（空质粒）及空白试剂分别转染大鼠源H9C2心肌细胞，并给予缺氧/复氧(H/R)干预。将心肌细胞分为空白组，空质粒组，DOCK1过表达组，空白+H/R组，空质粒+H/R组，DOCK1过表达+H/R组。用RT-PCR及Western bloting分别测定DOCK1mRNA及蛋白水平。流式细胞术、MTT分别测定细胞的凋亡率和增殖率。共聚焦激光扫描共焦显微镜观察心肌细胞骨架的改变。结果: DOCK1过表达组较空白组和空质粒组，DOCK1过表达+H/R组较空白+H/R组和空质粒+H/R组，人的DOCK1mRNA、DOCK1蛋白分别显著增加、心肌细胞凋亡率分别显著降低及增殖率分别显著增加。较DOCK1过表达组，，DOCK1过表达+H/R组人的DOCK1mRNA显著降低。较空白组、空质粒组及DOCK1过表达组，空白+H/R组、空质粒+H/R组及DOCK1过表达+H/R组大鼠的DOCK1mRNA、DOCK1蛋白分别显著降低、心肌细胞凋亡率分别显著增加及增殖率分别显著降低（所有P0.05）。结论: DOCK1可抑制细胞的凋亡，促进细胞骨架有序重排，促进细胞的增殖、存活；过表达DOCK1可抑制H/R诱导的心肌细胞凋亡增高、细胞骨架的排列紊乱、细胞增值、存活的降低。
[Abstract]:Objective: to investigate the effect of guanosine exchange factor DOCK1 on the survival of cardiac myocytes. Methods: rat H9C2 cardiomyocytes were transfected with recombinant eukaryotic plasmid pCXN2-Flag-hDOCK1 (human DOCK1 overexpression plasmid pCXN2-Flag1 (empty plasmid) and blank reagent respectively). The blank H / R group, The levels of DOCK1mRNA and protein were measured by RT-PCR and Western bloting. The apoptosis rate and proliferation rate were measured by flow cytometry. The cytoskeleton of cardiomyocytes was observed by confocal laser scanning confocal microscopy (confocal laser scanning confocal microscopy). Results: human DOCK1 mRNA-DOCK1 protein was significantly increased in DOCK1 overexpression group than that in blank H / R group and blank plasmid group. The apoptosis rate of cardiomyocytes was significantly decreased and the proliferation rate was significantly increased, compared with that of DOCK1 overexpression group, the human DOCK1mRNA was significantly decreased, compared with blank group, blank plasmid group and DOCK1 overexpression group, and blank H / R group. The expression of DOCK1 mRNA-DOCK1 protein in empty plasmid H / R group and DOCK1 overexpressed H / R group was significantly decreased, and the apoptosis rate and proliferation rate of cardiomyocytes were significantly increased (all P 0.05). Conclusion: DOCK1 can inhibit apoptosis, promote cytoskeleton rearrangement, promote cell proliferation and survival, and overexpression of DOCK1 can inhibit the increase of H / R induced cardiomyocyte apoptosis, the disorder of cytoskeleton arrangement, the increase of cell proliferation and the decrease of cell survival.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363

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