天津医药

天津医药 ›› 2018, Vol. 46 ›› Issue (3): 255-258.doi: 10.11958/20171207

• 实验研究 • 上一篇    下一篇

干扰SET8抑制血管平滑肌细胞转分化的研究

张东雪，高少辉，李同妙，周薇   

  1. 河北医科大学第四医院肾内科（邮编050011）
  • 收稿日期:2017-11-01 修回日期:2017-12-23 出版日期:2018-03-15 发布日期:2018-03-23
  • 通讯作者: 张东雪 E-mail:zhangdx009@163.com

The effect of interference SET8 on regulating the transdifferentiation of vascular smooth muscle cells

ZHANG Dong-xue, GAO Shao-hui, LI Tong-miao, ZHOU Wei   

  1. Departments of Nephrology, the Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China
  • Received:2017-11-01 Revised:2017-12-23 Published:2018-03-15 Online:2018-03-23

PDF (PC)

3408

摘要： 目的 探讨赖氨酸甲基转移酶 SET8 对大鼠血管平滑肌细胞（VSMCs）向成骨样细胞分化的影响。方法体外分离培养大鼠 VSMCs，将 VSMCs 随机分为正常对照组（未转染）、空质粒组（转染 NS-shRNA）、SET8-shRNA 组。应用反转录聚合酶链反应（RT-PCR）、蛋白免疫印迹（Western blot）法分别检测 SET8 和调节成骨与软骨分化的关键转录因子 RUNX2 mRNA 和蛋白的表达水平；应用酶联免疫法检测碱性磷酸酶（ALP）活性。结果 SET8-shRNA 可成功抑制 VSMCs 中 SET8 的表达；RT⁃PCR 及 Western blot 结果显示，SET8-shRNA 组 RUNX2 mRNA 和蛋白的表达较正常对照组和空质粒组明显下降（P＜0.05）。ALP 活性测定结果显示，SET8-siRNA 组 ALP 活性较正常对照组和 空质粒组显著降低（P＜0.05）。结论 干扰 SET8 基因表达能够抑制大鼠 VSMCs 向成骨样细胞转分化。

关键词: 肌细胞, 平滑肌, 细胞转分化, 组蛋白赖氨酸 N-甲基转移酶, RNA 干扰, 碱性磷酸酶, 赖氨酸甲基转移酶SET8, RUNX2

Abstract: Objective To investigate the effect of lysine methyltransferase SET8 on regulating cell transdifferentiation of rat vascular smooth muscle cells (VSMCs) into osteoblast-like cells. Methods VSMCs were obtained from rat thoracic aorta, and then randomly divided into control group (non-transfection), the empty plasmid group (transfect NS-shRNA) and SET8-shRNA group. The expression of SET8, runt⁃related transcription factor 2 (RUNX2) was detected by RT⁃PCR and Western blot assay. Alkaline phosphatase (ALP) activity was measured by enzyme linked immunoassay. Results The expression of SET8 in VSMCs was effectively inhibited by SET8-shRNA. RT-PCR and Western blot results showed that the expression of RUNX2 was decreased in cells after SET8-shRNA transfection (P＜0.05). ALP activity was significantly reduced after SET8-shRNA transfection (P＜0.05). Conclusion Interference with SET8 gene expression could inhibit the differentiation of VSMCs into osteo-like cells.

Key words: myocytes, smooth muscle, cell transdifferentiation, histone-lysine n-methyltransferase, RNA interference, alkaline phosphatase, lysine methyhransferase SET8, RUNX2

中图分类号: