慢病毒转染的β3-AR基因对心肌肥厚的影响

发布时间：2018-06-28 01:21

本文选题：β肾上腺素能受体 + 心肌细胞　；参考：《临床心血管病杂志》2017年01期

【摘要】：目的:探讨β3肾上腺素能受体(β3-AR)对SD大鼠乳鼠心肌肥大的影响及其机制。方法:体外培养SD大鼠乳鼠心肌细胞,用携带β3-AR基因的慢病毒转染细胞后,用去甲肾上腺素(NE)诱导细胞48h,建立心肌细胞肥大模型。实验分4组:空白对照组(Control组)、心肌肥厚组(NE组)、β3-AR基因转染+NE组(β3-AR组)、空病毒转染+NE组(空病毒组)。用免疫荧光法鉴定心肌细胞,倒置荧光显微镜观察病毒转染组绿色荧光蛋白(GFP)表达,免疫印迹法(Western Blot)检测β3-AR、丝裂原活化蛋白激酶p38(p38MAPK)、细胞外信号调控激酶(ERK1/2)、磷酸化p38MAPK(p-p38MAPK)和ERK(p-ERK1/2)及原癌基因c-myc、c-fos蛋白水平的表达。结果:(1)慢病毒介导β3-AR基因转染心肌细胞,β3-AR表达较空白对照组明显升高。(2)用Western Blot检测各实验组细胞原癌基因c-myc、c-fos表达,其中NE组、β3-AR组、空病毒组表达均高于空白对照组,其中β3-AR组cmyc、c-fos表达明显高于NE组。(3)NE组、β3-AR组、空病毒组p38MAPK及ERK1/2的磷酸化水平较空白对照组明显上调,其中β3-AR组表达最高。结论:慢病毒介导的β3-AR基因转染使心肌细胞有效高表达β3-AR,β3-AR可能通过MAPK通路促进心肌肥厚。
[Abstract]:Aim: to investigate the effect of 尾 3 adrenoceptor (尾 3 AR) on myocardial hypertrophy in neonatal SD rats and its mechanism. Methods: neonatal SD rat cardiomyocytes were cultured in vitro. The cells were transfected with lentivirus carrying 尾 3-AR gene and induced by norepinephrine (NE) for 48h to establish cardiomyocyte hypertrophy model. The experiment was divided into four groups: control group (blank control group), NE group (myocardial hypertrophy group), NE group (尾 3-AR group) transfected with 尾 3-AR gene, and NE group (empty virus group) transfected with empty virus. The expression of green fluorescent protein (GFP) in viral transfection group was observed by reverse fluorescence microscope. Western blot was used to detect the expression of 尾 3-AR, p38 MAPK, ERK1 / 2, phosphorylated p38 MAPK and ERK (p-ERK1 / 2) and proto-oncogene c-mycc-fos. Results: (1) the expression of 尾 _ 3-AR gene was significantly higher than that of the blank control group. (2) the expression of proto-oncogene c-mycnc-fos was detected by Western blot, and the expression of c-mycfs in NE group, 尾 _ 3-AR group and empty virus group was higher than that in blank control group. The expression of c-fos in 尾 3-AR group was significantly higher than that in NE group. (3) the phosphorylation levels of p38 MAPK and ERK1 / 2 in NE group, 尾 3-AR group and empty virus group were significantly higher than those in control group, and the highest expression was found in 尾 3-AR group. Conclusion: lentivirus-mediated 尾 _ 3-AR gene transfection can effectively overexpression 尾 _ 3-AR. 尾 _ 3-AR may promote myocardial hypertrophy through MAPK pathway.
【作者单位】：新疆石河子大学医学院第一附属医院心内二科;新疆医科大学第一附属医院心脏中心;
【基金】：国家自然科学基金-地区科学基金项目(No:81260028) 石河子大学科学技术研究发展计划“自然科学与技术创新”团队创新项目(No:2011ZRKXTD-07)
【分类号】：R541.6

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