MiR-125b-5p ameliorates ox-LDL-induced vascular endothelial cell dysfunction by negatively regulating TNFSF4/TLR4/NF-κB signaling
Abstract Background Oxidized low-density lipoprotein (ox-LDL)-induced endothelial cell dysfunction plays a crucial role in the progression of atherosclerosis (AS). Although miR-125b-5p is known to be involved in cardiovascular and cerebrovascular disorders, its function in ox-LDL-induced endothelial...
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BMC
2025-01-01
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| Series: | BMC Biotechnology |
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| Online Access: | https://doi.org/10.1186/s12896-025-00944-y |
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| author | Wenshuai He Limin Zhao Pengfei Wang Maojia Ren Yunfei Han |
| author_facet | Wenshuai He Limin Zhao Pengfei Wang Maojia Ren Yunfei Han |
| author_sort | Wenshuai He |
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| description | Abstract Background Oxidized low-density lipoprotein (ox-LDL)-induced endothelial cell dysfunction plays a crucial role in the progression of atherosclerosis (AS). Although miR-125b-5p is known to be involved in cardiovascular and cerebrovascular disorders, its function in ox-LDL-induced endothelial injury is still not well understood. Methods An in vitro AS cell model was established by exposing human umbilical vein endothelial cells (HUVECs) to 100 µg/mL ox-LDL for 24 h. A series of functional assays, including CCK-8 assay, flow cytometry, MDA and SOD kits, capillary-like network formation assay and ELISA assay were performed in vitro. TNFSF4/TLR4/NF-κB pathway-related protein expressions were measured by Western blot. Molecular mechanisms were elucidated through quantitative real-time PCR, western blot analysis, and luciferase reporter assays. Results Our investigation revealed that exposure to ox-LDL led to a downregulation in miR-125b-5p, while upregulating the expression of tumor necrosis factor (ligand) superfamily, member 4 (TNFSF4), TLR4, p-p65 and p-IkBa in HUVECs in a dose-dependent manner. We confirmed TNFSF4 as a direct target of miR-125b-5p. Ox-LDL exposure led to decreased cell viability and angiogenic capacity, along with increased apoptosis, inflammation, and oxidative stress in HUVECs. These effects were reversed by overexpressing miR-125b-5p or knocking down TNFSF4. Overexpression of TNFSF4 significantly reversed the effects brought about by miR-125b-5p in HUVECs exposed to ox-LDL. Moreover, miR-125b-5p inactivated the TLR4/NF-κB signaling pathway by negatively regulating TNFSF4. Conclusions In summary, our findings demonstrate that miR-125b-5p possessed an anti-inflammatory and anti-apoptosis against ox-LDL-induced HUVEC injury by regulating the TNFSF4/TLR4/NF-κB signaling, indicating that miR-125b-5p may have an important therapeutic function for AS. |
| format | Article |
| id | doaj-art-bfffca18a0524d4d8b0bcef1c7d44992 |
| institution | Kabale University |
| issn | 1472-6750 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | BMC |
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| series | BMC Biotechnology |
| spelling | doaj-art-bfffca18a0524d4d8b0bcef1c7d449922025-01-26T12:40:06ZengBMCBMC Biotechnology1472-67502025-01-0125111110.1186/s12896-025-00944-yMiR-125b-5p ameliorates ox-LDL-induced vascular endothelial cell dysfunction by negatively regulating TNFSF4/TLR4/NF-κB signalingWenshuai He0Limin Zhao1Pengfei Wang2Maojia Ren3Yunfei Han4Department of Emergency Medicine, Inner Mongolia People’s HospitalDepartment of Emergency Medicine, Inner Mongolia People’s HospitalDepartment of Emergency Medicine, Inner Mongolia People’s HospitalDepartment of Cardiology, Inner Mongolia People’s Hospital, Inner Mongolia AutonomousDepartment of Emergency Medicine, Inner Mongolia People’s HospitalAbstract Background Oxidized low-density lipoprotein (ox-LDL)-induced endothelial cell dysfunction plays a crucial role in the progression of atherosclerosis (AS). Although miR-125b-5p is known to be involved in cardiovascular and cerebrovascular disorders, its function in ox-LDL-induced endothelial injury is still not well understood. Methods An in vitro AS cell model was established by exposing human umbilical vein endothelial cells (HUVECs) to 100 µg/mL ox-LDL for 24 h. A series of functional assays, including CCK-8 assay, flow cytometry, MDA and SOD kits, capillary-like network formation assay and ELISA assay were performed in vitro. TNFSF4/TLR4/NF-κB pathway-related protein expressions were measured by Western blot. Molecular mechanisms were elucidated through quantitative real-time PCR, western blot analysis, and luciferase reporter assays. Results Our investigation revealed that exposure to ox-LDL led to a downregulation in miR-125b-5p, while upregulating the expression of tumor necrosis factor (ligand) superfamily, member 4 (TNFSF4), TLR4, p-p65 and p-IkBa in HUVECs in a dose-dependent manner. We confirmed TNFSF4 as a direct target of miR-125b-5p. Ox-LDL exposure led to decreased cell viability and angiogenic capacity, along with increased apoptosis, inflammation, and oxidative stress in HUVECs. These effects were reversed by overexpressing miR-125b-5p or knocking down TNFSF4. Overexpression of TNFSF4 significantly reversed the effects brought about by miR-125b-5p in HUVECs exposed to ox-LDL. Moreover, miR-125b-5p inactivated the TLR4/NF-κB signaling pathway by negatively regulating TNFSF4. Conclusions In summary, our findings demonstrate that miR-125b-5p possessed an anti-inflammatory and anti-apoptosis against ox-LDL-induced HUVEC injury by regulating the TNFSF4/TLR4/NF-κB signaling, indicating that miR-125b-5p may have an important therapeutic function for AS.https://doi.org/10.1186/s12896-025-00944-yAtherosclerosisHUVECsmiR-125b-5pTNFSF4TLR4/NF-κB signaling |
| spellingShingle | Wenshuai He Limin Zhao Pengfei Wang Maojia Ren Yunfei Han MiR-125b-5p ameliorates ox-LDL-induced vascular endothelial cell dysfunction by negatively regulating TNFSF4/TLR4/NF-κB signaling BMC Biotechnology Atherosclerosis HUVECs miR-125b-5p TNFSF4 TLR4/NF-κB signaling |
| title | MiR-125b-5p ameliorates ox-LDL-induced vascular endothelial cell dysfunction by negatively regulating TNFSF4/TLR4/NF-κB signaling |
| title_full | MiR-125b-5p ameliorates ox-LDL-induced vascular endothelial cell dysfunction by negatively regulating TNFSF4/TLR4/NF-κB signaling |
| title_fullStr | MiR-125b-5p ameliorates ox-LDL-induced vascular endothelial cell dysfunction by negatively regulating TNFSF4/TLR4/NF-κB signaling |
| title_full_unstemmed | MiR-125b-5p ameliorates ox-LDL-induced vascular endothelial cell dysfunction by negatively regulating TNFSF4/TLR4/NF-κB signaling |
| title_short | MiR-125b-5p ameliorates ox-LDL-induced vascular endothelial cell dysfunction by negatively regulating TNFSF4/TLR4/NF-κB signaling |
| title_sort | mir 125b 5p ameliorates ox ldl induced vascular endothelial cell dysfunction by negatively regulating tnfsf4 tlr4 nf κb signaling |
| topic | Atherosclerosis HUVECs miR-125b-5p TNFSF4 TLR4/NF-κB signaling |
| url | https://doi.org/10.1186/s12896-025-00944-y |
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