中国组织工程研究

中国组织工程研究 ›› 2014, Vol. 18 ›› Issue (42): 6752-6757.doi: 10.3969/j.issn.2095-4344.2014.42.006

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

人肺动脉平滑肌细胞中精氨酸酶Ⅱ与微小RNA-17的相互作用

靳有鹏，庞婷婷，王  伟，王玉林   

  1. 山东大学附属省立医院儿科，山东省济南市  250021
  • 修回日期:2014-09-11 出版日期:2014-10-08 发布日期:2014-10-08
  • 通讯作者: 王玉林，博士，教授，山东大学附属省立医院儿科，山东省济南市 250021
  • 作者简介:靳有鹏，女，1978年生，山东省济宁市人，汉族，2006年山东大学医学院毕业，博士，副教授，主要从事儿科重症医学及心血管疾病的研究。
  • 基金资助:

    山东省优秀中青年科学家科研奖励基金资助项目(BS 2011SW 040)

The interaction between arginase II and microRNA-17 in human pulmonary artery smooth muscle cells

Jin You-peng, Pang Ting-ting, Wang Wei, Wang Yu-lin   

  1. Department of Pediatrics, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China
  • Revised:2014-09-11 Online:2014-10-08 Published:2014-10-08
  • Contact: Wang Yu-lin, M.D., Professor, Department of Pediatrics, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China
  • About author:Jin You-peng, M.D., Associate professor, Department of Pediatrics, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China
  • Supported by:

    the Research Award Foundation for Outstanding Young Scientists of Shandong Province, No. BS 2011SW 040

PDF

411

被引次数

1

摘要：

背景：已有研究证实microRNA-17-92簇在肺动脉高压中发挥着重要作用，精氨酸酶Ⅱ又参与低氧诱导的肺动脉平滑肌细胞的增殖，而人肺动脉平滑肌细胞中微小RNA-17与精氨酸酶Ⅱ之间的相互作用尚未见研究报道。
目的：分析人肺动脉平滑肌细胞中，微小RNA-17与精氨酸酶Ⅱ的关系及其相互作用机制。
方法：选用4-8代的人肺动脉平滑肌细胞，分别给予转染微小RNA-17的抑制剂、增强剂、精氨酸酶Ⅱ小干扰RNA等处理，分别在常氧(体积分数21%O2)及低氧(体积分数1% O2)条件下培养。提取RNA和微小RNA，采用实时定量PCR法检测各组平滑肌细胞中微小RNA-17和精氨酸酶ⅡmRNA的表达，提取蛋白，采用Western blot法比较各组细胞中精氨酸酶Ⅱ等蛋白水平的表达。
结果与结论：低氧下人肺动脉平滑肌细胞中微小RNA-17及精氨酸酶Ⅱ的表达均明显增加，抑制微小RNA-17的表达可阻止低氧诱导的精氨酸酶Ⅱ的表达增加，微小RNA-17的过表达可使精氨酸酶Ⅱ表达上调，精氨酸酶Ⅱ基因敲除后，低氧诱导的微小RNA-17的表达受到抑制。提示人肺动脉平滑肌细胞中，精氨酸酶Ⅱ是微小RNA-17的一个新的靶基因，而且精氨酸酶Ⅱ可以反馈调节微小RNA-17的表达。



中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程


全文链接：

关键词: 组织构建, 组织工程, 肺动脉高压, 人肺动脉平滑肌细胞, 微小RNA-17, 精氨酸酶Ⅱ

Abstract:

BACKGROUND: microRNA-17 is confirmed to play an important role in the development of pulmonary hypertension. Some research has shown that hypoxia-induced proliferation in human pulmonary artery smooth muscle cell depends on the induction of arginase II. There is no report about whether there is some interaction between microRNA-17 and arginase II in human pulmonary artery smooth muscle cells.
OBJECTIVE: To investigate the possible interactions between microRNA-17 and arginase II in hypoxic human pulmonary artery smooth muscle cells.
METHODS: Passage 4 human pulmonary artery smooth muscle cells were cultured in 21% O2 and 5% CO2 (normoxia) or 1% O2 and 5% CO2 (hypoxia), and then transfected with mimic or inhibitor of microRNA-17 or arginase II-small interfering RNA. RNA, microRNA and protein were isolated separately. Expression of microRNA-17 and arginase II was detected with real-time quantitative PCR and western blot assay.
RESULTS AND CONCLUSION: The level of microRNA-17 was significantly increased in cultured human pulmonary artery smooth muscle cells exposed to 1% O2 hypoxia, as was arginase II mRNA and protein expression. Furthermore, inhibition of microRNA-17 expression decreased the mRNA and protein levels of arginase II in the human pulmonary artery smooth muscle cells under hypoxia. Conversely, over-expression of microRNA-17 increased the mRNA and protein levels of arginase II in the human pulmonary artery smooth muscle cells under normoxia and hypoxia. Knockdown of arginase II by siRNA abolished the hypoxia-induced up-regulation of microRNA-17 expression. These findings indicate that arginase II is a target gene of microRNA-17 and can regulate the expression of microRNA-17 in human pulmonary artery smooth muscle cells.



中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程


全文链接：

Key words: hypertension, pulmonary, myocytes, smooth muscle, microRNAs, arginase

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