天津医药

天津医药 ›› 2019, Vol. 47 ›› Issue (1): 23-26.doi: 10.11958/20181124

• 实验研究 • 上一篇    下一篇

P-糖蛋白过表达对INS-1和大鼠胰岛β细胞分化的影响

陈亚，高雅莉，李代清△   

  1. 基金项目：国家自然科学基金资助项目（81270891） 作者单位：天津医科大学代谢病医院，卫生部激素与发育重点实验室，天津市代谢性疾病重点实验室（邮编300070） 作者简介：陈亚（1993），女，硕士研究生在读，主要从事内分泌与代谢病相关研究 △通讯作者 E-mail: daiqingli68@126.com
  • 收稿日期:2018-07-23 修回日期:2018-11-20 出版日期:2019-01-15 发布日期:2019-01-15
  • 通讯作者: 李代清 E-mail:daiqingli68@126.com
  • 基金资助:
    国家自然科学基金;天津市科学委员会基金

Effects of P-glycoprotein overexpression on the differentiation of INS-1 and rat pancreatic β cells

CHEN Ya, GAO Ya-li, LI Dai-qing△   

  1. Key Laboratory of Hormones and Development (Ministry of Health), Tianjin Key Laboratory of Metabolic Diseases, Tianjin Metabolic Disease Hospital, Tianjin 300070, China △Corresponding Author E-mail：daiqingli68@126.com
  • Received:2018-07-23 Revised:2018-11-20 Published:2019-01-15 Online:2019-01-15
  • Contact: LI Daiqing E-mail:daiqingli68@126.com

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摘要： 目的 探讨P-糖蛋白过表达对大鼠胰岛β细胞系INS-1细胞和大鼠胰岛β细胞分化的影响。方法 分别 构建INS-1细胞（Ad-abcb1b腺病毒感染）和SD大鼠（Ins2-abcb1b随机插入）P-糖蛋白过表达模型。以阴性对照病毒 和野生型SD大鼠为对照组，P-糖蛋白过表达组和转基因SD大鼠为实验组，萃取细胞和胰岛总RNA，定量PCR分别 检测INS-1细胞和胰岛组织Abcb1b、Pdx1、Ins1、Ins2、Mafa、Mafb、Ngn3、Nkx6.1、Nkx2.2、Neurod1、Pax6等基因的表达。 结果 与对照组相比，P-糖蛋白过表达后，INS-1细胞Ngn3、Mafb转录水平升高（P＜0.05），Mafa转录水平降低（P＜ 0.001），其余基因表达差异无统计学意义。在野生型和转基因大鼠胰岛组织中，各基因转录水平差异均无统计学意 义（P＞0.05）。结论 P-糖蛋白过表达可能使INS-1细胞去分化，而对胰岛β细胞的分化无明显影响。

关键词: P糖蛋白, 胰岛素分泌细胞, 细胞分化, 糖尿病, 再生

Abstract: Objective To investigate the effect of P-glycoprotein overexpression on the differentiation of INS-1 cells and pancreatic β cells. Methods INS-1 cells were infected with adenovirus overexpressing P-glycoprotein. Ins2-abcb1b gene was randomly inserted into the rat islets, and P-glycoprotein overexpression model was successfully constructed. Empty adenovirus and wild-type SD rats were used as control groups. INS-1 cells and P-glycoprotein overexpression rats were used as experimental group. Total RNA was extracted both in INS-1 cells and rat islets. Transcription factors such as Abcb1b， Pdx1，Ins1，Ins2，Mafa，Mafb，Ngn3，Nkx6.1，Nkx2.2, Neurod1 and Pax6 were detected by real-time quantitative PCR. Results Compared with the control group，the transcription levels of Ngn3 and Mafb in INS-1 cells increased (P＜0.05), while Mafa decreased (P＜0.001). No significance was detected in other transcription factors. Unlike INS-1 cells, all of the transcription factors of islets were not affected (P＞0.05). Conclusion The overexpression of P-glycoprotein may dedifferentiate INS-1 cells, but has no effect on the differentiation of rat pancreatic beta cells.

Key words: P-glycoprotein, insulin-secreting cells, cell differentiation, diabetes mellitus, regeneration