Related Experiment Video
Updated: Nov 11, 2025
Using En Face Immunofluorescence Staining to Observe Vascular Endothelial Cells Directly
Published on: August 20, 2019
Kaempferol alleviates human endothelial cell injury through circNOL12/miR-6873-3p/FRS2 axis
Shuangzhan Li1, Meihua Hao1, Taisheng Wu1
1Department of Cardiology, Huaihe Hospital of Henan University, Kaifeng, Henan, China.
Insights
Kaempferol (Kae) reduces inflammation, oxidative stress, and apoptosis in endothelial cells by targeting the circNOL12/miR-6873-3p/FRS2 pathway. This study elucidates Kae's protective mechanisms against atherosclerosis-related cellular damage.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Atherosclerosis is a major cause of cardiovascular disease and stroke, driven by inflammation.
- Kaempferol (Kae) exhibits known pharmacological activities, but its precise mechanisms in endothelial cells are unclear.
- Understanding Kae's role in regulating inflammation, oxidative stress, and apoptosis in Human Umbilical Vein Endothelial Cells (HUVECs) is crucial.
Purpose of the Study:
- To investigate the detailed mechanisms by which Kaempferol (Kae) modulates inflammation, oxidative stress, and apoptosis in Human Umbilical Vein Endothelial Cells (HUVECs).
- To explore the role of the circNOL12/miR-6873-3p/FRS2 axis in Kaempferol's protective effects against oxidized low-density lipoprotein (ox-LDL) induced cellular damage.
Main Methods:
- Utilized real-time quantitative polymerase chain reaction (RT-qPCR) to assess gene and circRNA expression levels.
- Employed MTT assays for cell viability, flow cytometry for apoptosis quantification, and western blot for protein analysis.
- Investigated molecular interactions using dual-luciferase reporter and RNA pull-down assays to confirm the circNOL12/miR-6873-3p/FRS2 axis.
Main Results:
- Kaempferol (Kae) inhibited the upregulation of circNOL12 induced by oxidized low-density lipoprotein (ox-LDL) in HUVECs.
- Kae significantly attenuated ox-LDL-induced inflammation, oxidative stress (ROS, MDA levels), and apoptosis, effects reversed by circNOL12 overexpression.
- Identified miR-6873-3p as a target of circNOL12, with its upregulation counteracting circNOL12's effects, and FRS2 as negatively regulated by miR-6873-3p.
Conclusions:
- Kaempferol alleviates ox-LDL-induced inflammation, oxidative stress, and apoptosis in HUVECs.
- The protective effects of Kae are mediated through the regulation of the circNOL12/miR-6873-3p/FRS2 molecular axis.
- This study elucidates a novel molecular mechanism underlying Kaempferol's therapeutic potential in atherosclerosis.
Background:
Atherosclerosis, inflammatory disease, is a major reason for cardiovascular diseases and stroke. Kaempferol (Kae) has been well-documented to have pharmacological activities in the previous studies. However, the detailed mechanisms by which Kae regulates inflammation, oxidative stress, and apoptosis in Human Umbilical Vein Endothelial Cells (HUVECs) remain unknown.
Methods And Results:
The real-time quantitative polymerase chain reaction (RT-qPCR) was used to measure expression levels of circNOL12, nucleolar protein 12 (NOL12), miR-6873-3p, and Fibroblast growth factor receptor substrate 2 (FRS2) in HUVECs treated with either oxidized low-density lipoprotein (ox-LDL) alone or in combination with Kae. The cells viability was assessed by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-2H-tetrazol-3-ium bromide (MTT) assay. The inflammation and oxidative stress were assessed by checking inflammatory factors, Reactive Oxygen Species (ROS), Superoxide Dismutase (SOD), and Malondialdehyde (MDA) levels in ox-LDL-induced HUVECs. The apoptotic cells were quantified by flow cytometry assay. The western blot assay was used for measuring protein expression. The interaction relationship between miR-6873-3p and circNOL12 or FRS2 was analyzed by dual-luciferase reporter and RNA pull-down assays. Treatment with Kae could inhibit ox-LDL-induced the upregulation of circNOL12 in HUVECs. Importantly, Kae weakened ox-LDL-induced inflammation, oxidative stress, and apoptosis in HUVECs, which was abolished by overexpression of circNOL12. What's more, miR-6873-3p was a target of circNOL12 in HUVECs, and the upregulation of miR-6873-3p overturned circNOL12 overexpression-induced effects on HUVECs treated with ox-LDL and Kae. FRS2 was negatively regulated by miR-6873-3p in HUVECs.
Conclusion:
Kae alleviated ox-LDL-induced inflammation, oxidative stress, and apoptosis in HUVECs by regulating circNOL12/miR-6873-3p/FRS2 axis.
