基础医学与临床 ›› 2010, Vol. 30 ›› Issue (8): 852-856.

• 研究论文 • 上一篇    下一篇

高糖上调大鼠主动脉平滑肌细胞骨桥蛋白表达

揭伟 罗泊涛 姜汉国 陈锦 申志华   

  1. 广东医学院 基础学院 广东医学院 基础学院
  • 收稿日期:2009-10-26 修回日期:2009-12-04 出版日期:2010-08-05 发布日期:2010-08-05
  • 通讯作者: 申志华

High glucose-induced up-regulation of osteopontin in rat aortic vascular smooth muscle cells

Wei JIE, Bo-tao LUO, Han-guo JIANG, Jin CHEN, Zhi-hua SHEN   

  1. College of Basic Sciences; Guangdong Medical College College of Basic Sciences; Guangdong Medical College
  • Received:2009-10-26 Revised:2009-12-04 Online:2010-08-05 Published:2010-08-05
  • Contact: Zhi-hua SHEN

摘要: 目的 探讨高糖对大鼠血管平滑肌细胞(VSMCs)骨桥蛋白(OPN)表达的影响及机制。方法 SD大鼠腹腔注射链脲佐菌素复制Ⅰ型糖尿病模型,免疫组化、RT-PCR和western blot检测大鼠主动脉OPN和PCNA的表达;大鼠主动脉VSMCs经含正常糖(5.5mmol/L)、高糖(25mmol/L)、甘露醇(25mmol/L)、高糖+GF109203X(蛋白激酶C抑制剂,10μmol/L),高糖+ NAC(N-acetyl-l-cysteine,NF-κB抑制剂,10μmol/L)的DMEM处理48 h,免疫荧光及western blot检测OPN的表达,western blot 分析PCNA、PKC和 p65的表达。结果 高糖血症大鼠主动脉VSMCs高表达OPN及PCNA。原代VSMCs经高糖刺激后OPN、PCNA、PKC及p65表达明显增加;GF109203X和NAC均可下调PCNA的高表达,但仅GF109203X抑制高糖诱导的OPN高表达。结论 高糖可能通过激活PKC信号诱导OPN表达并促进VSMCs增殖从而在糖尿病性血管并发症中起作用。

关键词: 血管平滑肌细胞, 高糖, Osteopontin, PKC, NF-κB

Abstract: Objective To investigate the effects and mechanism of high glucose on the osteopontin (OPN) expression in rat aortic vascular smooth muscle cells (VSMCs). Methods SD rats were intraperitoneally injected with streptozotocin, OPN and PCNA expression in rat thoracic aortas were detected by immunohistochemical staining, RT-PCR and western blot analysis; After primary rat aortic VSMCs treated with DMEM medium supplemented with normal glucose (5.5mmol/L), high glucose (25mmol/L), mannitol (25mmol/L), high glucose + GF109203X (PKC inhibitor, 10μmol/L) and high glucose + NAC (N-acetyl-l-cysteine, NF-κB inhibitor, 10μmol/L) for 48 h, OPN expression was examined by immunofluorescence and western blot, expression of PCNA, PKC, and p65 were assessed by western blot analysis. Results OPN and PCNA expression were increased in hyperglycemic rat aortic VSMCs. Expression of OPN, PCNA, PKC, and p65 were significantly up-regulated in VSMCs exposed to high glucose. Expression of PCNA was attenuated by GF109203X and NAC in high glucose-treated VSMCs, while the expression of OPN was only inhibited by GF109203X. Conclusion High glucose induces OPN expression and cell proliferation in VSMCs may be due to the activation of PKC signal, which play a critical role in diabetic vascular complications.

Key words: vascular smooth muscle cells, high glucose, osteopontin, PKC, NF-κB