scholarly journals CRISPR/dCas9 Transcriptional Activation of Endogenous Apolipoprotein AI and Paraoxonase 1 in Enterocytes Alleviates Endothelial Cell Dysfunction

Biomolecules ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 1769
Author(s):  
Laura Toma ◽  
Teodora Barbălată ◽  
Gabriela M. Sanda ◽  
Loredan S. Niculescu ◽  
Anca V. Sima ◽  
...  
Keyword(s):  
Protein Expression ◽  
Transcriptional Activation ◽  
Sirtuin 1 ◽  
Paraoxonase 1 ◽  
High Density Lipoproteins ◽  
Peroxisome Proliferator ◽  
Apolipoprotein Ai ◽  
Monocyte Chemoattractant Protein 1 ◽  
Inflammatory Status ◽  
Gene And Protein Expression

Atherosclerosis is the main cause of cardiovascular diseases with high prevalence worldwide. A promising therapeutic strategy to reverse atherosclerotic process is to improve the athero-protective potential of high-density lipoproteins (HDL). Since the small intestine is a source of HDL, we aimed to activate transcription of the endogenous HDL major proteins, apolipoprotein AI (ApoAI) and paraoxonase 1 (PON1), in enterocytes, and to evaluate their potential to correct the pro-inflammatory status of endothelial cells (EC). Caco-2 enterocytes were transfected with CRISPR activation plasmids targeting ApoAI or PON1, and their gene and protein expression were measured in cells and conditioned medium (CM). ATP binding cassette A1 and G8 transporters (ABCA1, ABCG8), scavenger receptor BI (SR-BI), and transcription regulators peroxisome proliferator-activated receptor γ (PPARγ), liver X receptors (LXRs), and sirtuin-1 (SIRT1) were assessed. Anti-inflammatory effects of CM from transfected enterocytes were estimated through its ability to inhibit tumor necrosis factor α (TNFα) activation of EC. Transcriptional activation of ApoAI or PON1 in enterocytes induces: (i) increase of their gene and protein expression, and secretion in CM; (ii) stimulation of ABCA1/G8 and SR-BI; (iii) upregulation of PPARγ, LXRs, and SIRT1. CM from transfected enterocytes attenuated the TNFα-induced inflammatory and oxidative stress in EC, by decreasing TNF receptor 1, monocyte chemoattractant protein-1, and p22phox. In conclusion, transcriptional activation of endogenous ApoAI or PON1 in enterocytes by CRISPR/dCas9 system is a realistic approach to stimulate biogenesis and function of major HDL proteins which can regulate cholesterol efflux transporters and reduce the inflammatory stress in activated EC.

scholarly journals Expression levels of MCP-1, HGF, and IGF-1 in endometriotic patients compared with non-endometriotic controls

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Sahel Heidari ◽  
Roya Kolahdouz-Mohammadi ◽  
Sepideh Khodaverdi ◽  
Nader Tajik ◽  
Ali-Akbar Delbandi
Keyword(s):  
Gene Expression ◽  
Growth Factor ◽  
Protein Expression ◽  
Mononuclear Cells ◽  
Endometrial Stromal Cells ◽  
Peripheral Blood Mononuclear ◽  
Monocyte Chemoattractant Protein 1 ◽  
Gene And Protein Expression ◽  
Its Gene ◽  
Hepatocyte Growth

Abstract Background To study the concentrations of monocyte chemoattractant protein-1 (MCP-1), hepatocyte growth factor (HGF), and insulin-like growth factor-1 (IGF-1) in peritoneal fluid (PF) and serum, and to evaluate their expressions by PF and peripheral blood mononuclear cells (PFMCs and PBMCs, respectively), and ectopic and eutopic endometrial stromal cells of patients with endometriosis (EESCs and EuESCs, respectively) compared with controls. Methods The concentrations of mentioned cytokines in serum and PF, as well as their expression in PBMCs, PFMCs, EuESCs and EESCs from endometriosis patients and controls were assessed. Results The levels of MCP-1, HGF, and IGF-1 in serum and PF in women with endometriosis were significantly higher than the controls (P < 0.05–P < 0.001). Gene expression of MCP-1 and IGF-1 in the PFMCs, PBMCs and EESCs also showed an increased level compared to controls (P < 0.05–P < 0.01). The protein expression of MCP-1 and IGF-1 by PFMCs was statistically higher in endometriotic women (P < 0.05 and P < 0.01, respectively). The gene and protein expression of HGF in PFMCs and its gene expression by EESCs were significantly higher in endometriotic women compared to controls (P < 0.05–P < 0.01). Conclusions The higher concentrations of mentioned cytokines in serum and PF and their higher expression by PFMCs and EESCs in endometriosis patients may contribute to the development of endometriosis.

scholarly journals Metabolic regulation of exercise-induced angiogenesis

2019 ◽  
Vol 1 (1) ◽  
pp. H1-H8 ◽  
Author(s):  
Tatiane Gorski ◽  
Katrien De Bock
Keyword(s):  
Skeletal Muscle ◽  
Transcriptional Activation ◽  
Metabolic Regulation ◽  
Molecular Mechanisms ◽  
Metabolic Reprogramming ◽  
Sirtuin 1 ◽  
Peroxisome Proliferator ◽  
Valuable Insight ◽  
Peroxisome Proliferator Activated Receptor ◽  
Exercise Induced

Skeletal muscle relies on an ingenious network of blood vessels, which ensures optimal oxygen and nutrient supply. An increase in muscle vascularization is an early adaptive event to exercise training, but the cellular and molecular mechanisms underlying exercise-induced blood vessel formation are not completely clear. In this review, we provide a concise overview on how exercise-induced alterations in muscle metabolism can evoke metabolic changes in endothelial cells (ECs) that drive muscle angiogenesis. In skeletal muscle, angiogenesis can occur via sprouting and splitting angiogenesis and is dependent on vascular endothelial growth factor (VEGF) signaling. In the resting muscle, VEGF levels are controlled by the estrogen-related receptor γ (ERRγ). Upon exercise, the transcriptional coactivator peroxisome-proliferator-activated receptor-γ coactivator-1α (PGC1α) orchestrates several adaptations to endurance exercise within muscle fibers and simultaneously promotes transcriptional activation of Vegf expression and increased muscle capillary density. While ECs are highly glycolytic and change their metabolism during sprouting angiogenesis in development and disease, a similar role for EC metabolism in exercise-induced angiogenesis in skeletal muscle remains to be elucidated. Nonetheless, recent studies have illustrated the importance of endothelial hydrogen sulfide and sirtuin 1 (SIRT1) activity for exercise-induced angiogenesis, suggesting that EC metabolic reprogramming may be fundamental in this process. We hypothesize that the exercise-induced angiogenic response can also be modulated by metabolic crosstalk between muscle and the endothelium. Defining the underlying molecular mechanisms responsible for skeletal muscle angiogenesis in response to exercise will yield valuable insight into metabolic regulation as well as the determinants of exercise performance.

scholarly journals Enhanced pan-peroxisome proliferator-activated receptor gene and protein expression in adipose tissue of diet-induced obese mice treated with telmisartan

2014 ◽  
Vol 99 (12) ◽  
pp. 1663-1678 ◽  
Author(s):  
Aline Penna-de-Carvalho ◽  
Francielle Graus-Nunes ◽  
Júlia Rabelo-Andrade ◽  
Carlos Alberto Mandarim-de-Lacerda ◽  
Vanessa Souza-Mello
Keyword(s):  
Adipose Tissue ◽  
Protein Expression ◽  
Receptor Gene ◽  
Peroxisome Proliferator ◽  
Obese Mice ◽  
Peroxisome Proliferator Activated Receptor ◽  
Gene And Protein Expression

scholarly journals AMP-Activated Protein Kinase Abundance and Activation is increased with the β-Adrenergic agonist Zilpaterol Hydrochloride in Muscle from Feedlot Cattle

2021 ◽  
Vol 2 (2) ◽  
pp. 1-6
Author(s):  
Bradley Joseph Johnson
Keyword(s):  
Protein Kinase ◽  
Protein Expression ◽  
Muscle Tissue ◽  
Fiber Type ◽  
Feedlot Cattle ◽  
Mrna Abundance ◽  
Peroxisome Proliferator ◽  
Gene And Protein Expression ◽  
Zilpaterol Hydrochloride ◽  
Igf I

The objective of this research was to evaluate the effect of Zilpaterol Hydrochloride (ZH) on myogenic or adipogenic gene and protein expression in skeletal muscle. Two feeding trials and one cell culture experiments were conducted. Semimembranosus muscle tissue was collected from steers that had been fed a diet containing 8.3 mg of ZH/kg DM for the last 0 or 20 d of the finishing period with a 3 d withdrawal period. To test the mode of action an in vitromodel was used with, isolated bovine satellite cells isolated from muscle tissue. Real Time-QPCR (RTQPCR) was used to measure the relative mRNA abundance of Adenosine Monophosphate Protein Kinase α (AMPK), Myosin Heavy Chain (MHC) I, MHCIIA, MHCIIX, Insulin-like Growth Factor I (IGF-I), β-adrenergic receptor (βAR) 1 and 2, peroxisome proliferator-activated receptor gamma (PPARγ), and Stearoyl-CoA Desaturase (SCD). Western blotting was used to measure the relative protein abundance of AMPK and Phosphorylated-AMPK (pAMPK). No differences were detected in relative mRNA abundance of AMPK, MHCIIA, IGF-I, βAR1 and βAR2. However, MHCI, SCD, and PPARγ mRNA expression was decreased and MHCIIX mRNA increased from ZH fed cattle compared to non-ZH. For one experiment, AMPK protein expression increased, while in another experiment, AMPK phosphorylation increased with ZH fed animals. The increase in MHCIIX mRNA with ZH fed cattle indicated the start of a fiber type shift towards larger diameter fibers. This shift may have been due to increased expression and phosphorylation of AMPK. These data suggest that the shift increase in MHCIIX was likely due to the ZH administration.

scholarly journals l-Theanine Ameliorates d-Galactose-Induced Brain Damage in Rats via Inhibiting AGE Formation and Regulating Sirtuin1 and BDNF Signaling Pathways

2021 ◽  
Vol 2021 ◽  
pp. 1-13
Author(s):  
Li Zeng ◽  
Ling Lin ◽  
Ling Chen ◽  
Wenjun Xiao ◽  
Zhihua Gong
Keyword(s):  
Protein Expression ◽  
Signaling Pathways ◽  
Hippocampal Neurons ◽  
Nuclear Factor Κb ◽  
Peroxisome Proliferator ◽  
Inflammatory Factor ◽  
Peroxisome Proliferator Activated Receptor ◽  
Gene And Protein Expression ◽  
Mrna And Protein Expression ◽  
Age Receptors

The maintenance of homeostasis is essential for mitigating stress and delaying degenerative diseases such as Alzheimer’s disease (AD). AD is generally defined as the abnormal production of β-amyloid (Aβ) and advanced glycation end products (AGEs). The effects of l-theanine on Aβ and AGE generation were investigated in this study. Decreased AGEs and Aβ1-42 levels were reflected by increased acetylcholine (ACh) concentration and acetylcholinesterase (AChE) activity inhibition compared to model rats. l-Theanine also inhibited nuclear factor-κB (p65) protein expression by activating sirtuin1 (SIRT1), reducing inflammatory factor expression, and downregulating the mRNA and protein expression of AGE receptors (RAGE). Superoxide dismutase 2 and catalase protein expressions were markedly upregulated by l-theanine, whereas oxidative stress-related injury was alleviated. The expression of peroxisome proliferator-activated receptor-γ coactivator 1α (PGC-1α) was also found to be increased. H&E staining showed that the apoptosis of hippocampal neurons was mitigated by decreased Bax and cleaved-caspase-3 protein expression and the increase of Bcl-2 protein expression. Moreover, l-theanine increased the gene and protein expression of brain-derived neurotrophic factor (BDNF). These findings suggest that the potential preventive effects of l-theanine against AD may be attributed to its regulation of SIRT1 and BDNF proteins and its mitigation of AGEs/RAGE signaling pathways in the brain tissue of AD model rats.

scholarly journals p-Cymene Modulate Oxidative Stress and Inflammation in Murine Macrophages: Potential Implication in Atherosclerosis

2020 ◽  
Vol 18 (2) ◽  
pp. 151-157
Author(s):  
Tong Wu ◽  
Zahra Mazhar ◽  
Dhuha Alsayrafi ◽  
Mahdi Garelnabi
Keyword(s):  
Oxidative Stress ◽  
Antioxidant Properties ◽  
Paraoxonase 1 ◽  
High Density Lipoproteins ◽  
Necrosis Factor Alpha ◽  
Potential Implication ◽  
Monocyte Chemoattractant Protein 1 ◽  
Tnf Α ◽  
Factor Alpha ◽  
Dose Dependent

Introduction: p-Cymene (p-CYM) is a common chemical used in air fresheners. Objective: The study was designed to investigate the molecular effect of p-CYM on macrophages. Materials and Methods: Macrophages (RAW 264.7) were treated with p-CYM (50 uM/L, 150 uM/L and 250 uM/L) for 6 hours, and 24 hours). Gene involved in inflammation, such as the Tumor Necrosis Factor-alpha (TNF-α), and the Monocyte Chemoattractant Protein-1 (MCP-1) and other genes known for their antioxidant activity such as the Paraoxonase 1 (PON-1) were analyzed. Results: Cells treated with p-CYM have shown 30% up-regulation of MCP-1 after 24 hour of exposure; and also a differential up-regulation of TNF-α. However, treatment with p-CYM has resulted in a considerable (37%) dose-dependent downregulation of PON-1 after 24 hours of exposure. PON-1 is known for its antioxidant properties protecting High-Density Lipoproteins (HDL) from oxidation. Conclusion:: Our findings demonstrate that exposure to p-CYM over time promotes oxidative stress by downregulating antioxidants genes as shown in PON-1 and also stimulates inflammation, a key process during the initiation and progression of atherosclerosis.

scholarly journals Gene and Protein Expression Profile of Selected Molecular Targets Mediating Electrophysiological Function in Pgc-1α Deficient Murine Atria

2018 ◽  
Vol 19 (11) ◽  
pp. 3450
Author(s):  
Karan Chadda ◽  
Charlotte Edling ◽  
Haseeb Valli ◽  
Shiraz Ahmad ◽  
Christopher Huang ◽  
...  
Keyword(s):  
Protein Expression ◽  
Expression Profile ◽  
Expression Profiles ◽  
Molecular Targets ◽  
Cholinergic Receptor ◽  
Na Channel ◽  
Peroxisome Proliferator ◽  
Peroxisome Proliferator Activated Receptor ◽  
Receptor Modulation ◽  
Gene And Protein Expression

Increases in the prevalence of obesity, insulin resistance, and metabolic syndrome has led to the increase of atrial fibrillation (AF) cases in the developed world. These AF risk factors are associated with mitochondrial dysfunction, previously modelled using peroxisome proliferator activated receptor-γ (PPARγ) coactivator-1 (Pgc-1)-deficient murine cardiac models. We explored gene and protein expression profiles of selected molecular targets related to electrophysiological function in murine Pgc-1α−/− atria. qPCR analysis surveyed genes related to Na+-K+-ATPase, K+ conductance, hyperpolarisation-activated cyclic nucleotide-gated (Hcn), Na+ channels, Ca2+ channels, and indicators for adrenergic and cholinergic receptor modulation. Western blot analysis for molecular targets specific to conduction velocity (Nav1.5 channel and gap junctions) was performed. Transcription profiles revealed downregulation of molecules related to Na+-K+-ATPase transport, Hcn-dependent pacemaker function, Na+ channel-dependent action potential activation and propagation, Ca2+ current generation, calsequestrin-2 dependent Ca2+ homeostasis, and adrenergic α1D dependent protection from hypertrophic change. Nav1.5 channel protein expression but not gap junction expression was reduced in Pgc-1α−/− atria compared to WT. Nav1.5 reduction reflects corresponding reduction in its gene expression profile. These changes, as well as the underlying Pgc-1α−/− alteration, suggest potential pharmacological targets directed towards either upstream PGC-1 signalling mechanisms or downstream ion channel changes.

scholarly journals PPAR γ/Nnat/NF-κB Axis Involved in Promoting Effects of Adiponectin on Preadipocyte Differentiation

2019 ◽  
Vol 2019 ◽  
pp. 1-9 ◽  
Author(s):  
Wenkai Yang ◽  
Wenjin Yuan ◽  
Xinghua Peng ◽  
Meiling Wang ◽  
Jie Xiao ◽  
...  
Keyword(s):  
Protein Expression ◽  
Peroxisome Proliferator ◽  
Preadipocyte Differentiation ◽  
Peroxisome Proliferator Activated Receptor ◽  
Monocyte Chemoattractant Protein 1 ◽  
Expression Levels ◽  
Real Time Quantitative Pcr ◽  
Inflammatory Conditions ◽  
Ppar Γ ◽  
Total Antioxidant

A previous study has demonstrated that adiponectin (APN) could promote preadipocyte differentiation, and the present study further explored its mechanism. 3T3-L1 cells were infected with adenovirus holding human adiponectin gene apM1 and mouse neuronatin (Nnat) shRNA and initiated differentiation while coculturing with mature adipocytes stimulated with LPS. After 8 days, preadipocyte differentiation was observed by Oil Red O staining. Real-time quantitative PCR was used to evaluate mRNA expression levels of monocyte chemoattractant protein-1 (MCP-1), interleukin- (IL-) 6, IL-8, and tumor necrosis factor α (TNF-α). The levels of reactive oxygen species (ROS), total antioxidant capacity (T-AOC), malondialdehyde (MDA), and superoxide dismutase (SOD) in 3T3-L1 cells were detected. Western blotting was done to quantify the protein expression levels of Nnat, peroxisome proliferator-activated receptor (PPAR) γ, p65, and inhibitor of nuclear factor κB (IκB) α. Results demonstrated that APN overexpression markedly increased preadipocyte differentiation; inhibited gene expression of MCP-1, IL-6, IL-8, and TNF-α; reduced ROS and MDA release; increased T-AOC and SOD levels; upregulated Nnat, PPAR γ, and IκB α protein expressions; and downregulated p65 protein expression under LPS stimulation. However, the effects of APN were markedly attenuated when Nnat expression was knocked down. Taken together, the present study provided evidences that the effects of APN on promoting preadipocyte differentiation under inflammatory conditions via anti-inflammation and antioxidative stress may be regulated by the PPAR γ/Nnat/NF-κB signaling pathway.

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