天津医药

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棕榈酸对小鼠胰岛和INS-1E细胞作用的研究

张艳玲,邹本良   

  1. 中国中医科学院西苑医院
  • 收稿日期:2011-08-01 修回日期:2012-03-08 出版日期:2012-08-15 发布日期:2012-08-15
  • 通讯作者: 张艳玲

The chronic effect of palmitate on the function of isolated mouse islets and clonal B cell INS-1E cells

  • Received:2011-08-01 Revised:2012-03-08 Published:2012-08-15 Online:2012-08-15
  • Contact: YANLING ZHANG

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摘要： 目的 探讨高脂对INS-1E细胞和小鼠胰岛的慢性作用。方法 雌性NMRI小鼠,6～10周齡,苯巴比妥腹腔麻醉,應用膠原酶技術消化胰腺分離胰島。传代培养的INS-1E细胞和分离的小鼠胰岛分别在含0、0.4 mmol/L棕榈酸的RPMI1640中培养72 h，然后在含3.3、16.7 mmol/L葡萄糖的Krebs-Ringer缓冲液中培养60 min，留取上清液行胰岛素测定。INS-1E细胞在含不同浓度的棕榈酸 的RPMI1640中培养72 h，提取其总RNA，合成相应的cDNA，再行RT-PCR检测Pdx1，胰岛素1，胰岛素2和葡萄糖转运子2的基因表达。结果：高脂培养后INS-1E细胞和小鼠胰岛的基础胰岛素分泌增加，糖刺激的胰岛素分泌减少，INS-1E细胞的胰岛素1，胰岛素2和葡萄糖转运子2的mRNA水平下降。结论：高脂显示了对胰岛B细胞的慢性毒性作用。

关键词: 脂肪酸毒性, B细胞, 基因

Abstract: Objective To explore the chronic effect of excess palmitate on INS-1E cells and isolated mouse islets. Methods Islets were isolated by the collagenase digestion technique from adult NMRI mice. Both passaged INS-1E cells and isolated mouse islets were incubated in RPMI 1640 supplemented with 0 or 0.4 mmol/L palmitate for 72 h, respectively. After preincubation, both INS-1E cells and islets were incubated in Krebs-Ringer buffer containing 3.3 or 16.7 mmol/L glucose for 1 h, respectively. Correspondingly upper medium was taken for insulin assay. Total RNA was extracted from INS-1E cells after 72 h incubation with 0 or 0.4 mmol/L palmitate, cDNA was synthesized, Pdx1, Insulin 1, Insulin 2, and GLUT2 mRNA expressions were tested in INS-1E cells by RT-PCR. Results Basal insulin secretion was increased, and glucose-stimulated insulin secretion was impaired in both INS-1E cells and isolated mouse islets after long-term excess palmitate incubation, and insulin 1, insulin 2, and GLUT2 mRNA levels were diminished in INS-1E cells. Conclusion Long-term exposure to higher level of palmitate concentration may cause dysfunction of pancreatic islet cells.

Key words: lipotoxicity, B cell, gene