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CPT1a Knockdown reduces CPT1a, TGF-b1, a-SMA, NLRP3 expression induced by 7-KC in NRK-52E renal epithelial cells and increases lipid content.
Source publication
Background:
Dyslipidemia causes renal damage; however, the detailed molecular mechanism has not been clarified. It is known that carnitine palmitoyl transferase 1-a (CPT1a) gene encodes an enzyme involved in fatty acid oxidation and, therefore, lipid content. In the present study, we investigated whether the accumulation of lipids induced by 7-ket...
Contexts in source publication
Context 1
... CPT1a knockdown in 7-ketocholesterol treated cells inhibits the profibrotic and NLRP3 inducing effects of 7-ketocholesterol As shown in Fig. 3A, CPT1a knockdown provokes a decrease in CPT1a expression in all the 7-KC treated groups, revealing the efficiency of the silencing ...
Context 2
... shown in Fig. 3B, analysis of intracellular lipid accumulation indicated the increased accumulation of lipid droplets in all the groups that underwent CPT1a ...
Context 3
... has been found that CPT1a activation promotes NLRP3 inflammasome activation in bone marrow derived macrophages [26]. Our results are in line with this observation, since increases in CPT1a expression are accompanied by increases in NLRP3 expression and silencing or decreasing CPT1a by C75 pre-treatment, decreases NLRP3 inflammasome expression (Figs. 3 and ...
Citations
... For in vitro experiments, the macrophages were treated for 5 days with COM dissolved in DMEM at 25, 50, 100, and 200 μg/ml; as a control, the cells were cultured under normal conditions. C75 pre-treatment was administrated at 10 μg/ml for 2 h before COM treatment [27]. ...
Objective and design
Kidney stones commonly occur with a 50% recurrence rate within 5 years, and can elevate the risk of chronic kidney disease. Macrophage-to-myofibroblast transition (MMT) is a newly discovered mechanism that leads to progressive fibrosis in different forms of kidney disease. In this study, we aimed to investigate the role of MMT in renal fibrosis in glyoxylate-induced kidney stone mice and the mechanism by which signal transducer and activator of transcription 6 (STAT6) regulates MMT.
Methods
We collected non-functioning kidneys from patients with stones, established glyoxylate-induced calcium oxalate stone mice model and treated AS1517499 every other day in the treatment group, and constructed a STAT6-knockout RAW264.7 cell line. We first screened the enrichment pathway of the model by transcriptome sequencing; detected renal injury and fibrosis by hematoxylin eosin staining, Von Kossa staining and Sirius red staining; detected MMT levels by multiplexed immunofluorescence and flow cytometry; and verified the binding site of STAT6 at the PPARα promoter by chromatin immunoprecipitation. Fatty acid oxidation (FAO) and fibrosis-related genes were detected by western blot and real-time quantitative polymerase chain reaction.
Results
In this study, we found that FAO was downregulated, macrophages converted to myofibroblasts, and STAT6 expression was elevated in stone patients and glyoxylate-induced kidney stone mice. The promotion of FAO in macrophages attenuated MMT and upregulated fibrosis-related genes induced by calcium oxalate treatment. Further, inhibition of peroxisome proliferator-activated receptor-α (PPARα) eliminated the effect of STAT6 deletion on FAO and fibrosis-associated protein expression. Pharmacological inhibition of STAT6 also prevented the development of renal injury, lipid accumulation, MMT, and renal fibrosis. Mechanistically, STAT6 transcriptionally represses PPARα and FAO through cis-inducible elements located in the promoter region of the gene, thereby promoting MMT and renal fibrosis.
Conclusions
These findings establish a role for STAT6 in kidney stone injury-induced renal fibrosis, and suggest that STAT6 may be a therapeutic target for progressive renal fibrosis in patients with nephrolithiasis.
... Carnitine palmitoyltransferase 1 (CPT1) has three types, namely CPT1A, CPT1B and CPT1C, with different properties and tissue distributions 18 . CP-T1A is enriched in the liver and exerts a key role in the β-oxidation of fatty acids [18][19][20] . In brown fat cells cultured in vitro, CPT1A is capable of improving β-oxidation and mitochondrial function of fatty acids 20 . ...
Objective:
The purpose of this study was to elucidate the regulatory role of microRNA-324-5p (miRNA-324-5p) in inhibiting inflammatory response of diabetic vessels by regulating CPT1A level, thus alleviating the development of type 2 diabetes mellitus (T2DM).
Patients and methods:
Arterial vessels (splenic artery) and serum exosomes were extracted from 30 T2DM patients and 30 non-T2DM subjects treated in Binzhou People's Hospital from 2015 to 2019. Relative levels of miRNA-324-5p and CPT1A in each subject were detected. Then, VSMCs were induced with high-glucose, followed by detection of inflammatory factor levels. Next, the regulatory effects of miRNA-324-5p and CPT1A on viability, 5-Ethynyl-2'-deoxyuridine (EdU)-positive ratio, and release of inflammatory factors in VSMCs were determined. Finally, Dual-Luciferase reporter assay was conducted to verify the interaction between miRNA-324-5p and CPT1A.
Results:
The results revealed that compared with non-T2DM subjects, miRNA-324-5p was downregulated in splenic arteries and exosomes in T2DM patients. High-glucose treatment in VSMCs triggered the release of the inflammatory factors. In addition, the overexpression of miRNA-324-5p in VSMCs reduced viability and inflammatory factor levels, and the inhibited trends were partially reversed by overexpression of CPT1A. CPT1A was indicated to be the target gene binding miRNA-324-5p.
Conclusions:
MiRNA-324-5p exerts an inhibitory effect on T2DM-induced inflammation in blood vessels by negatively regulating CPT1A level and reducing the release of inflammatory factors. MiRNA-324-5p might be a promising therapeutic target for T2DM.
Kidney tubular epithelial cells (TECs) have a crucial role in the damage and repair response to acute and chronic injury. To adequately respond to constant changes in the environment, TECs have considerable bioenergetic needs, which are supported by metabolic pathways. Although little is known about TEC metabolism, a number of ground-breaking studies have shown that defective glucose metabolism or fatty acid oxidation in the kidney has a key role in the response to kidney injury. Imbalanced use of these metabolic pathways can predispose TECs to apoptosis and dedifferentiation, and contribute to lipotoxicity and kidney injury. The accumulation of lipids and aberrant metabolic adaptations of TECs during kidney disease can also be driven by receptors of the innate immune system. Similar to their actions in innate immune cells, pattern recognition receptors regulate the metabolic rewiring of TECs, causing cellular dysfunction and lipid accumulation. TECs should therefore be considered a specialized cell type - like cells of the innate immune system - that is subject to regulation by immunometabolism. Targeting energy metabolism in TECs could represent a strategy for metabolically reprogramming the kidney and promoting kidney repair.
Various liver diseases caused by liver damage seriously affect people's health. The purpose of this study was to clarify that the effects and mechanism of Carnitine palmitoyltransferase 1 (Cpt1a) on oxidative stress and inflammation in liver injury. It was found that the expression of Cpt1a mRNA was up-regulated in model mice of liver injury. So, over-expression of Cpt1a increased reactive oxygen species (ROS) production and malondialdehyde (MDA) levels, and reduced superoxide dismutase (SOD), glutathione (GSH) and glutathione peroxidase (GSH-px) levels in vitro model of liver injury. It was also shown that over-expression of Cpt1a suppressed the Nuclear factor-erythroid-2-related factor 2 (Nrf2)/ heme oxygenase-1 (HO-1) signaling pathway. In summary, these data indicate that Cpt1a promotes ROS-induced oxidative stress in liver injury via the Nrf2/HO-1 and nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome signaling pathway.
