分泌型卷曲相关蛋白1通过Wnt/β-catenin通路抑制血管平滑肌细胞钙化
SFRP1 Inhibits Vascular Smooth Muscle Cell Calcification Via the Wnt/β-catenin Signaling Pathway
- 中山大学学报（医学科学版） 2025年46卷第5期 页码：816-825
- 作者机构：
1.南方医科大学珠江医院心脏中心实验室心血管内科，广东 广州 510280
2.南方医科大学珠江医院转化医学研究中心，广东 广州 510280
- 作者简介：
罗文君，第一作者，研究方向：血管钙化，E-mail：april99728@163.com
- 基金信息：广东省自然科学基金(2021A1515011097)
DOI：10.13471/j.cnki.j.sun.yat-sen.univ（med.sci）.2025.0511
中图分类号： R543收稿：2025-06-26，
修回：2025-08-04，
录用：2025-08-17，
纸质出版：2025-09-20
- 稿件说明：
移动端阅览
罗文君,刘灿昭,王先宝.分泌型卷曲相关蛋白1通过Wnt/β-catenin通路抑制血管平滑肌细胞钙化[J].中山大学学报（医学科学版）,2025,46(05):816-825.
LUO Wenjun,LIU Canzhao,WANG Xianbao.SFRP1 Inhibits Vascular Smooth Muscle Cell Calcification Via the Wnt/β-catenin Signaling Pathway[J].Journal of Sun Yat-sen University(Medical Sciences),2025,46(05):816-825.
罗文君,刘灿昭,王先宝.分泌型卷曲相关蛋白1通过Wnt/β-catenin通路抑制血管平滑肌细胞钙化[J].中山大学学报（医学科学版）,2025,46(05):816-825. DOI： 10.13471/j.cnki.j.sun.yat-sen.univ（med.sci）.2025.0511.
LUO Wenjun,LIU Canzhao,WANG Xianbao.SFRP1 Inhibits Vascular Smooth Muscle Cell Calcification Via the Wnt/β-catenin Signaling Pathway[J].Journal of Sun Yat-sen University(Medical Sciences),2025,46(05):816-825. DOI： 10.13471/j.cnki.j.sun.yat-sen.univ（med.sci）.2025.0511.
摘要
目的
2
探讨分泌型卷曲相关蛋白1（SFRP1）通过Wnt/β-catenin通路对血管平滑肌细胞成骨分化的影响。
方法
2
使用原代人主动脉血管平滑肌细胞，转染SFRP1小干扰RNA（siSFRP1）敲低SFRP1在人血管平滑肌细胞表达，转染慢病毒Lenti-Sfrp1过表达SFRP1，使用3 mmol/L的磷酸二氢钠（Pi）刺激构建细胞血管钙化模型。体内实验采用维生素D
3
（VD
3
）构建急性血管钙化模型，腹腔注射SFRP1抑制剂WAY316606（0.5 mg/kg，7 d），Micro-CT检测小鼠主动脉钙化程度。Western blot检测成骨分化指标RUNX2、BMP2，血管平滑肌细胞收缩标志SM22α，Wnt信号通路β-catenin蛋白水平表达。钙含量检测与茜素红染色评估血管钙化严重程度，研究SFRP1对血管平滑肌钙化的影响。
结果
2
在原代人血管平滑肌细胞钙化模型与急性血管钙化模型中，SFRP1蛋白水平表达明显下调（0.30
<math id="M1"><mo>±</mo></math>
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203151&type=
2.28600001
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203157&type=
1.52400005
0.02，
P
=0.02；0.15
<math id="M2"><mo>±</mo></math>
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203151&type=
2.28600001
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203157&type=
1.52400005
0.03，
P
=0.04）。使用小分子干扰RNA敲低SFRP1，明显上调RUNX2表达（2.91
<math id="M3"><mo>±</mo></math>
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203151&type=
2.28600001
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203157&type=
1.52400005
0.38，
P
＜0.05），下调SM22α表达（0.48
<math id="M4"><mo>±</mo></math>
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203151&type=
2.28600001
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203157&type=
1.52400005
0.08，
P
＜0.05），同时激活Wnt/β-catenin信号通路（2.01
<math id="M5"><mo>±</mo></math>
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203151&type=
2.28600001
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203157&type=
1.52400005
0.11，
P
=0.003）。体内实验使用SFRP1抑制剂加重小鼠主动脉钙化，上调BMP2与β-catenin表达（3.11
<math id="M6"><mo>±</mo></math>
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203151&type=
2.28600001
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203157&type=
1.52400005
0.55，
P
=0.04；3.97
<math id="M7"><mo>±</mo></math>
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203151&type=
2.28600001
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203157&type=
1.52400005
0.44，
P
=0.03）。使用慢病毒过表达SFRP1，明显下调RUNX2（1.34
<math id="M8"><mo>±</mo></math>
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203151&type=
2.28600001
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203157&type=
1.52400005
0.04，
P
=0.02）与β-catenin表达（1.06
<math id="M9"><mo>±</mo></math>
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203151&type=
2.28600001
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203157&type=
1.52400005
0.06，
P
=0.04），上调SM22α表达（0.74
<math id="M10"><mo>±</mo></math>
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203151&type=
2.28600001
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203157&type=
1.52400005
0.03，
P
=0.03）。茜素红染色定量与钙含量检测显示过表达SFRP1抑制血管平滑肌细胞钙化（1.36
<math id="M11"><mo>±</mo></math>
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203151&type=
2.28600001
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203157&type=
1.52400005
0.08，
P
=0.000 6；1.51
<math id="M12"><mo>±</mo></math>
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203151&type=
2.28600001
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203157&type=
1.52400005
0.03，
P
=0.002 1）。
结论
2
SFRP1通过抑制Wnt/β-catenin通路减轻血管平滑肌细胞成骨分化。
Abstract
Objective
2
To investigate the effect of secreted frizzled related protein 1（SFRP1） via Wnt/β-catenin signaling pathway on calcification of vascular smooth muscle cell.
Methods
2
Primary human aortic vascular smooth muscle cells were transfected with SFRP1-specific small interfering RNA （siSFRP1） to knock down SFRP1 expression， transfected with lentiviral vector Lenti-Sfrp1 to overexpress SFRP1， and stimulated with 3 mmol/L sodium dihydrogen phosphate （Pi） to establish a cellular vascular calcification model.
In vivo
， a mouse model of acute vascular calcification was established using vitamin D
3
（VD
3
） treatment. The SFRP1 inhibitor WAY-316606 was administered intraperitoneally （0.5 mg/kg daily for 7 days）. Aortic calcification was quantified by micro-computed tomography （micro-CT）. Western blot analysis was performed to detect the expression of osteogenic differentiation markers （RUNX2 and BMP2）， vascular smooth muscle cell contractile marker SM22α， and β-catenin protein in the Wnt signaling pathway. The severity of vascular calcification was evaluated through calcium content measurement and Alizarin Red staining. These approaches were employed to investigate the effect of SFRP1 on VSMC calcification.
Results
2
In the primary human vascular smooth muscle cell calcification model and the acute vascular calcification model， the expression of SFRP1 protein was significantly do
wn-regulated （0.30
<math id="M13"><mo>±</mo></math>
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203151&type=
2.28600001
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203157&type=
1.52400005
0.02，
P
= 0.02； 0.15
<math id="M14"><mo>±</mo></math>
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203151&type=
2.28600001
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203157&type=
1.52400005
0.03，
P
= 0.04）. The expression of RUNX2 （2.91
<math id="M15"><mo>±</mo></math>
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203151&type=
2.28600001
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203157&type=
1.52400005
0.38，
P
＜ 0.05） was significantly up-regulated and SM22α （0.48
<math id="M16"><mo>±</mo></math>
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203151&type=
2.28600001
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203157&type=
1.52400005
0.08，
P
＜ 0.05） were significantly down-regulated by small interfering RNA （siRNA）， whereas the Wnt/β-catenin signaling pathway was also significantly activated （2.01
<math id="M17"><mo>±</mo></math>
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203151&type=
2.28600001
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203157&type=
1.52400005
0.11，
P
= 0.003）.
In vivo
， inhibition of SFRP1 exacerbated aortic calcification and significantly up-regulated the expression of BMP2 （3.11
<math id="M18"><mo>±</mo></math>
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203151&type=
2.28600001
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203157&type=
1.52400005
0.55，
P
= 0.04） and β-catenin （3.97
<math id="M19"><mo>±</mo></math>
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203151&type=
2.28600001
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203157&type=
1.52400005
0.44，
P
= 0.03）. Lentiviral overexpression of SFRP1 significantly downregulated RUNX2 （1.34
<math id="M20"><mo>±</mo></math>
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203152&type=
2.28600001
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203168&type=
1.52400005
0.04，
P
= 0.02） and β-catenin （1.06
<math id="M21"><mo>±</mo></math>
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203152&type=
2.28600001
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203168&type=
1.52400005
0.06，
P
= 0.04） expression， while upregulating SM22α （0.74
<math id="M22"><mo>±</mo></math>
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203152&type=
2.28600001
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203168&type=
1.52400005
0.03，
P
= 0.03）. Quantitative alizarin red staining and calcium content assays demonstrated that SFRP1 overexpression significantly inhibited vascular smooth muscle cell calcification （1.36
<math id="M23"><mo>±</mo></math>
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203151&type=
2.28600001
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203157&type=
1.52400005
0.08，
P
=0.000 6；1.51
<math id="M24"><mo>±</mo></math>
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203158&type=
2.28600001
https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=95203183&type=
1.52400005
0.03，
P
=0.002 1）.
Conclusion
2
SFRP1 attenuates vascular smooth muscle cell calcification via the Wnt/β-catenin signaling pathway.
关键词
Keywords
references
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