Matrine inhibits inflammatory response induced by TNF-α in human umbilical vein endothelial cells through miR-25-3p-mediated Klf4 pathway.
10.19540/j.cnki.cjcmm.20230510.709
- Author:
Zi-Ping XIANG
1
;
Yan-Jie LI
1
;
Huan MA
2
;
Xing WANG
2
;
Hui-Xin ZHANG
3
;
Chao WANG
2
Author Information
1. Graduate School, North China University of Science and Technology Tangshan 063210, China Key Laboratory of Metabolic Disease, Hebei General Hospital Shijiazhuang 050051, China.
2. Key Laboratory of Metabolic Disease, Hebei General Hospital Shijiazhuang 050051, China.
3. Chemical Engineering Institute, Shijiazhuang University Shijiazhuang 050035, China.
- Publication Type:Journal Article
- Keywords:
Krüppel like transcription factor 4;
human umbilical vein endothelial cells;
inflammatory response;
matrine;
miR-25-3p
- MeSH:
Humans;
Tumor Necrosis Factor-alpha/metabolism*;
MicroRNAs/metabolism*;
Human Umbilical Vein Endothelial Cells;
Matrines;
Interleukin-6/genetics*;
Signal Transduction;
Antagomirs;
Inflammation/metabolism*;
Luciferases/pharmacology*;
RNA, Messenger;
Apoptosis
- From:
China Journal of Chinese Materia Medica
2023;48(17):4731-4737
- CountryChina
- Language:Chinese
-
Abstract:
This study aimed to analyze the effect of matrine on tumor necrosis factor-α(TNF-α)-induced inflammatory response in human umbilical vein endothelial cells(HUVECs) and explore whether the underlying mechanism was related to the miR-25-3p-mediated Krüppel-like factor 4(Klf4) pathway. The HUVEC cell inflammation model was induced by TNF-α stimulation. After 24 or 48 hours of incubation with different concentrations of matrine(0.625, 1.25, and 2.5 mmol·L~(-1)), CCK-8 assay was used to detect cell proliferation. After treatment with 2.5 mmol·L~(-1) matrine for 48 h, the expression of TNF-α, interleukin-6(IL-6), interleukin-1β(IL-1β), and Klf4 mRNA and miR-25-3p was detected by real-time fluorescence-based quantitative PCR, and the protein expression of TNF-α, IL-6, IL-1β, and Klf4 was detected by Western blot. The anti-miR-25-3p was transfected into HUVECs, and the effect of anti-miR-25-3p on TNF-α-induced cell proliferation and inflammatory factors was detected by the above method. The cells were further transfected with miR-25-3p and incubated with matrine to detect the changes in proliferation and expression of related inflammatory factors, miR-25-3p, and Klf4. The targeting relationship between miR-25-3p and Klf4 was verified by bioinformatics analysis and dual luciferase reporter gene assay. The results displayed that matrine could inhibit TNF-α-induced HUVEC proliferation, decrease the mRNA and protein expression of TNF-α, IL-6, and IL-1β, increase the mRNA and protein expression of Klf4, and reduce the expression of miR-25-3p. Bioinformatics analysis showed that there were specific complementary binding sites between miR-25-3p and Klf4 sequences. Dual luciferase reporter gene assay confirmed that miR-25-3p negatively regulated Klf4 expression in HUVECs by targeting. The inhibition of miR-25-3p expression can reduce TNF-α-induced cell proliferation and mRNA and protein expression of TNF-α, IL-6, and IL-1β. MiR-25-3p overexpression could reverse the effect of matrine on TNF-α-induced cell proliferation and the mRNA and protein expression of TNF-α, IL-6, IL-1β, and Klf4. This study shows that matrine inhibits the inflammatory response induced by TNF-α in HUVECs through miR-25-3p-mediated Klf4 pathway.
