Pigment epithelium-derived factor alleviates endothelial injury by inhibiting Wnt/β-catenin pathway.
BACKGROUND
Oxidized low-density lipoprotein (ox-LDL) can induce endothelial injury and plays a vital role in the procession and development of atherosclerosis. Little is known regarding whether Wnt/β-catenin pathway is involved in ox-LDL-induced endothelial injury or whether it further promotes atherosclerosis via increased oxidative stress. This study aimed to investigate the role of Wnt/β-catenin pathway in ox-LDL-induced vascular endothelial injury and determine whether pigment epithelium-derived factor (PEDF) could alleviate ox-LDL-induced endothelial injury by inhibiting Wnt/β-catenin pathway.


METHODS
Injury of human umbilical vein endothelial cells (HUVECs) was evaluated with an MTT assay, by monitoring lactate dehydrogenase (LDH) release and determining the apoptotic ratio. The expression of β-catenin (non-phosphorylated-β-catenin), disheveled-1 (Dvl-1) and Cyclin D1 was analyzed with western blotting and quantitative real-time PCR. Oxidative stress status was assessed by measuring the levels of reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD) and nitric oxide (NO).


RESULTS
Exposure of HUVECs to ox-LDL led to a decrease in cell viability and an increase in LDH release and apoptosis with concomitant enhancement of oxidative stress, as assessed by increased ROS and MDA generation, as well as decreased SOD activity and NO levels. Similar to lithium chloride (LiCl, a Wnt/β-catenin pathway activator), ox-LDL up-regulated the expression of β-catenin, Dvl-1 and Cyclin D1, markers of Wnt/β-catenin pathway activation. However, ox-LDL-induced activation of Wnt/β-catenin pathway, as well as ox-LDL-induced cell injury and oxidative stress, were synergistically promoted by LiCl and mitigated by Dickkopf 1 (DKK-1), an inhibitor of Wnt/β-catenin pathway. Additionally, ox-LDL-induced HUVEC injury and apoptosis, oxidative stress and activation of Wnt/β-catenin pathway were suppressed by PEDF, while they were further strengthened by a small interfering RNA of PEDF.


CONCLUSIONS
Wnt/β-catenin pathway may mediate ox-LDL-induced endothelial injury via oxidative stress, and PEDF ameliorates endothelial injury by suppressing Wnt/β-catenin pathway and subsequently reducing oxidative stress.
Department of Geriatric Cardiology, Chinese PLA General Hospital, Beijing, 100853, China.
Shouyuan
S
Ma
Key Laboratory of Atherosclerosis in Universities of Shandong and Institute of Atherosclerosis, Taishan Medical University, Taian, Shandong, 271000, China.|College of Basic Medical Sciences, Taishan Medical University, Taian, Shandong, 271000, China.
Shutong
S
Yao
Key Laboratory of Atherosclerosis in Universities of Shandong and Institute of Atherosclerosis, Taishan Medical University, Taian, Shandong, 271000, China.
Hua
H
Tian
Key Laboratory of Atherosclerosis in Universities of Shandong and Institute of Atherosclerosis, Taishan Medical University, Taian, Shandong, 271000, China.
Peng
P
Jiao
Key Laboratory of Atherosclerosis in Universities of Shandong and Institute of Atherosclerosis, Taishan Medical University, Taian, Shandong, 271000, China.
Nana
N
Yang
Department of Geriatric Cardiology, Chinese PLA General Hospital, Beijing, 100853, China. zhuping301@139.com.
Ping
P
Zhu
Key Laboratory of Atherosclerosis in Universities of Shandong and Institute of Atherosclerosis, Taishan Medical University, Taian, Shandong, 271000, China. shucunqin@hotmail.com.
Shucun
S
Qin
D050197:Atherosclerosis; D042783:Endothelial Cells / Q000378:metabolism* / Q000473:pathology; D005136:Eye Proteins; D006801:Humans; D009414:Nerve Growth Factors; D018384:Oxidative Stress; D015843:Serpins; D014471:Umbilical Veins / Q000166:cytology / Q000378:metabolism / Q000473:pathology; D060449:Wnt Signaling Pathway; D000099174:Pigment Epithelium-Derived Factor
D005136:Eye Proteins; D009414:Nerve Growth Factors; D015843:Serpins; D000099174:Pigment Epithelium-Derived Factor
Atherosclerosis; Endothelial injury; Oxidative stress; Oxidized low-density lipoprotein; Pigment epithelium-derived factor; Wnt/β-catenin pathway