KLF5/LINC00346/miR‑148a‑3p axis regulates inflammation and endothelial cell injury in atherosclerosis

Abstract

Atherosclerosis (AS) is the main pathological basis of cardiovascular diseases, which are related to high morbidity and mortality rates. The present study aimed to investigate the role of the Krüppel-like factor 5 (KLF5)/LINc00346/miR-148a-3p loop in AS. The expression levels of KLF5 in serum and of KLF5/LINc00346/miR-148a-3p in human umbilical vein endothelial cells (HUVEcs) were detected by RT-qPcR analysis. The protein expression levels of KLF5, phosphorylated (p-)endothelial nitric oxide synthase (eNOS) and eNOS in HUVEcs were analyzed by western blot analysis. changes in the levels of TNF-α, IL-1β, IL-6 and nitric oxide (NO) were determined in the supernatant through the application of available commercial kits. The binding of KLF5 to the promoter region of LINC00346 was verified by chromatin immunoprecipitation (chIP)-PcR assay. The combinatory interaction between KLF5 and LINc00346, LINc00346 and miR-148a-3p, and miR-148a-3p and KLF5 was confirmed by luciferase reporter assay. The results revealed that KLF5 expression was increased in the serum of patients with AS and also in oxidized low-density lipoprotein (OX-LdL)-stimulated HUVEcs. The transcription factor KLF5 promoted the transcription of LINc00346. KLF5 interference or LINc00346 interference inhibited the expression of inflammatory factors and functional injury in OX-LdL-stimulated HUVEcs. LINc00346 functioned as a sponge of miR-148a-3p. miR-148a-3p overexpression inhibited the expression of inflammatory factors and functional injury in OX-LdL-stimulated HUVEcs and miR-148a-3p targeted KLF5 expression. On the whole, the present study demonstrates that KLF5 interference induces the downregulation of LINC00346 and also inhibits inflammation and functional injury in OX OX-LdL-stimulated HUVEcs by upregulating miR-148a-3p expression.FANGFANG WANG, JIYONG GE, SHENGLAN HUANG, cHANGLE ZHOU, ZIKAI SUN, YING SONG, YAJING XU and YUAN JI department of cardiovascular disease, changzhou No.

MiR-148a-3p attenuates apoptosis and inflammation by targeting CNTN4 in atherosclerosis

“…The THP-1 macrophages were then induced by 100 μg/mL oxLDL into foam cell formation. It has been reported that miR-148a-3p suppressed the expression of inflammatory factors in oxLDL-induced human umbilical vein endothelial cells (HUVECs) via targeting KLF ( 29 ). Here, we revealed that miR-148a-3p was markedly decreased and CNTN4 expression level was markedly increased in the THP-1 + PMA + oxLDL group, and the overexpression of miR-148-3p remarkably increased the expression of CNTN4 in oxLDL-induced THP-1 macrophages.…”

“…A total of 7 miRNAs, including 5 AS is a lipid-driven inflammatory disease, and inflammation has long been considered a marker of AS. Recent studies have found that miR-148a-3p plays a critical role in inflammatory diseases (27)(28)(29) and angiogenesis (30,31). The knockdown of SULT2B1b has been shown to inhibit the inflammatory response via increasing the miR-148a-3p level in macrophages (32).…”

RNA sequencing-based identification of the regulatory mechanism of microRNAs, transcription factors, and corresponding target genes involved in vascular dementia

“…These ten dysregulated TFs were found in VaD for the first time. Klf5 , the most downregulated TF, is an active factor associated with miRNAs in cancer and cardiovascular disease ( Drosatos et al, 2016 ; Yang et al, 2018 ; Nan et al, 2021 ; Wang Y. et al, 2021 ). Klf5 was significantly downregulated in the blood of rats with ischemic stroke obtained from the GSE21136.…”

Participation of Krüppel-like Factors in Atherogenesis

“…KLF5 can contribute to endothelial dysfunction by regulating the expression of genes involved in vascular tone, permeability, and inflammation. For example, KLF5 can increase the expression of endothelin-1, a potent vasoconstrictor [ 74 , 75 ], and decrease the expression of eNOS, which produces the vasodilator nitric oxide [ 76 , 77 ]. KLF5 can also increase the expression of vascular cell adhesion molecule 1 (VCAM-1), C-X-C Motif Chemokine Ligand 1 (CXCL1), matrix metallopeptidase 9 (MMP9), VEGF, and other genes [ 78 , 79 , 80 ] that directly play a key role in many processes, including leukocyte adhesion and migration, resulting in increased vascular permeability and inflammation.…”

“…KLF5 can promote inflammation by regulating the expression of genes involved in the innate immune response [ 20 , 77 ]. KLF5 can increase the expression of cytokines such as IL-1, IL-6, and TNF-α and adhesion molecules.…”