天津医药

天津医药 ›› 2020, Vol. 48 ›› Issue (5): 380-384.doi: 10.11958/20193488

• 实验研究 • 上一篇    下一篇

骨保护素参与高磷诱导大鼠血管平滑肌细胞钙化调节的机制初探 #br#

窦建新 1，孙丽萍 1，刘长山 1，王秀军 1，明义 1，苏亮 2△
  

  1. 1山东省潍坊市人民医院内分泌科（邮编261000），2心内二科
  • 收稿日期:2019-11-18 修回日期:2020-01-17 出版日期:2020-05-15 发布日期:2020-06-24
  • 通讯作者: 苏亮 E-mail:suliang15963699766@163.com
  • 基金资助:
    山东省优秀中青年科学家科研奖励基金（2014BSA07014）；山东省医药卫生科技发展计划项目（2016WS0658）

The mechanism of osteoprotegerin involved in the calcification of vascular smooth muscle cells induced by β-glycerophosphate #br#

DOU Jian-xin1, SUN Li-ping1, LIU Chang-shan1, WANG Xiu-jun1, MING Yi1, SU Liang2△ #br#   

  1. 1 Department of Endocrinology, 2 Department of Cardiology Ⅱ, Weifang People's Hospital,Weifang 261000, China
  • Received:2019-11-18 Revised:2020-01-17 Published:2020-05-15 Online:2020-06-24

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摘要： 摘要：目的 以β-磷酸甘油诱导大鼠血管平滑肌细胞（VSMC）钙化模型为研究对象，初步探讨骨保护素（OPG）干
预对VSMC钙化的影响及机制。方法 体外分离培养大鼠主动脉VSMC。PBS为对照组，以10 mmol/L β-磷酸甘油诱
导钙化10 d为钙化对照组，β-磷酸甘油诱导钙化同时予以剂量梯度（1、4、8 μg/L）的OPG干预为OPG干预组。茜素红
染色检测各组钙化结节形成情况。邻甲酚酞络合酮比色法检测细胞钙沉积含量。比色法测定碱性磷酸酶（ALP）活性。
免疫蛋白印迹（Western blot）检测各组Notch1/RBP-JK通路蛋白的表达。结果 茜素红染色显示，与钙化对照组比较，
OPG干预组大鼠VSMC钙化结节明显减少，OPG干预组的钙含量、ALP活性均明显下降（均P＜0.05），且有剂量依赖性。
Western blot结果显示，与钙化对照组比较，OPG干预组的Notch1、RBP-JK、Jagged1、Msx2蛋白表达水平明显下降（均P＜
0.05）。结论 OPG可能通过下调Notch1/RBP-JK通路蛋白表达，参与大鼠VSMC钙化过程调节。

关键词: 骨保护素；肌细胞, 平滑肌；血管钙化；受体, Notch1；Notch1/RBP-JK通路

Abstract:

Abstract：Objective To investigate the mechanism of osteoprotegerin intervention on thecalcification of vascular
smooth muscle cells (VSMC) induced by β -glycerophosphate in rats. Methods Rat aortic smooth muscle cells were
isolated and cultured in vitro. Cells treated with PBS were used as the control group. Cells treated with 10 mmol/L β -
glycerophosphate for 10 days to induce the calcification were used as the calcified control group, while cells treated with β-
glycerophosphate and osteoprotegerin (OPG) in dose gradients (1, 4 and 8 μg/L) were used as the osteoprotegerin intervention
group. Alizarin red staining was used to detect the formation of calcified nodules. The content of calcium deposition was
measured by the o-cresolphthalein complexone method. The alkaline phosphatase activity was determined by the
colorimetric method. The protein expressions of Notch1/RBP-JK pathway were detected by Western blot assay. Results
Alizarin red staining showed that compared with the calcified control group, the calcified nodules of VSMCs were
significantly reduced in OPG intervention groups. Compared with calcification control group, the calcium content and ALP
activity decreased significantly in OPG intervention group in a dose-dependent manner (P < 0.05). In addition, compared
with calcification control group, the expression levels of Notch 1, RBP-JK, Jagged1 and Msx2 protein decreased significantly
in OPG intervention group (P < 0.05). Conclusion Osteoprotegerin is involved in the regulation of vascular calcification by
the down regulating the expression of Notch1/RBP-JK pathway protein.

Key words: osteoprotegerin, myocytes, smooth muscle, vascular calcification, receptor, Notch1, Notch1/RBP-JK
pathway