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IRGM/Irgm1 regulates macrophage autophagy in response to ox-LDL in vitro is an early-stage event. (A-F) THP-1 cells were transfected with either control or IRGM plasmids (Lanes 1, 2) or transduced with either control or IRGM siRNA (Lanes 3, 4) and then stimulated with ox-LDL (50 µg/mL) for 3 h. The WB (A) and Qpcr (C) were used to detect the transfection efficiency and quantitative analysis results are shown (n = 3) (B). Quantitative data represent the fold change after normalised to GAPDH. (D-F) The expression of autophagy-related proteins LC3 and P62 were detected by western blot. (G) After stimulating THP-1 cell with ox-LDL (50 μg/mL) for 3 h, the autophagy double-labelled adenovirus fluorescent probe was used to detect the expression of autophagosomes (yellow) and autophagolysosomes (red). Green fluorescence was quenched in an acidic environment. Scale bar: 10 µm. (I, J) Quantitative analysis of the yellow and red puncta in (G). (H) After stimulating THP-1 cell with ox-LDL (50 μg/mL) for 48 h, the autophagy double-labelled adenovirus fluorescent probe was used to detect the expression of autophagosomes (yellow) and autophagolysosomes (red). Green fluorescence is quenched in an acidic environment. Scale bar: 10 µm. (K, L) Quantitative analysis of yellow and red puncta in (H). *p < 0.05, **p < 0.01, ****p < 0.0001, #p < 0.05, ###p < 0.001. Results are presented as mean ± SD. Statistical analysis: unpaired Student's t-test.
Source publication
Rationale: Atherosclerosis plaque rupture (PR) is the pathological basis and chief culprit of most acute cardiovascular events and death. Given the complex and important role of macrophage apoptosis and autophagy in affecting plaque stability, an important unanswered question include is whether, and how, immunity-related GTPase family M protein (IR...
Contexts in source publication
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... the important role of macrophage survival in PR, we further hypothesized that IRGM/Irgm1 might regulate macrophage autophagy. First, IRGM/IRGM1 expression were modified by specific siRNA (knockdown) or IRGM plasmid (knock-in) ( Figure 3A-C, Figure S3A-C). Western blot results showed that after ox-LDL (50 μg/mL) treatment for 3 h, compared with control, overexpression of IRGM significantly increased LC3II/I and decreased p62. ...
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... the important role of macrophage survival in PR, we further hypothesized that IRGM/Irgm1 might regulate macrophage autophagy. First, IRGM/IRGM1 expression were modified by specific siRNA (knockdown) or IRGM plasmid (knock-in) ( Figure 3A-C, Figure S3A-C). Western blot results showed that after ox-LDL (50 μg/mL) treatment for 3 h, compared with control, overexpression of IRGM significantly increased LC3II/I and decreased p62. ...
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... blot results showed that after ox-LDL (50 μg/mL) treatment for 3 h, compared with control, overexpression of IRGM significantly increased LC3II/I and decreased p62. In contrast, knocking down IRGM/Irgm1 significantly decreased LC3 II/I and increased p62 ( Figure 3D-F, Figure S3E, F). Meanwhile, knocking down Irgm1 significantly reduced mRNA levels of autophagy-related genes Atg5, Atg7, Beclin1, and LC3 ( Figure S3D). ...
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... blot results showed that after ox-LDL (50 μg/mL) treatment for 3 h, compared with control, overexpression of IRGM significantly increased LC3II/I and decreased p62. In contrast, knocking down IRGM/Irgm1 significantly decreased LC3 II/I and increased p62 ( Figure 3D-F, Figure S3E, F). Meanwhile, knocking down Irgm1 significantly reduced mRNA levels of autophagy-related genes Atg5, Atg7, Beclin1, and LC3 ( Figure S3D). ...
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... contrast, knocking down IRGM/Irgm1 significantly decreased LC3 II/I and increased p62 ( Figure 3D-F, Figure S3E, F). Meanwhile, knocking down Irgm1 significantly reduced mRNA levels of autophagy-related genes Atg5, Atg7, Beclin1, and LC3 ( Figure S3D). Furthermore, we used mRFP-GFP tandem fluorescently labelled LC3II to evaluate autophagosome-lysosome fusion. ...
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... puncta indicate fusion. Compared with control, after ox-LDL treatment for 3 h, overexpression of IRGM significantly increased RFPpositive/GFP-negative spots, while knocking down IRGM/Irgm1 significantly reduced RFP-positive/GFPnegative spots ( Figure 3G ...
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... results of caspase3/9 activities were also detected in THP-1 cells ( Figure 4D, E). In addition, after ox-LDL treatment for 48 h, knocking down Irgm1 in RAW264.7 cells also significantly decreased the expression of cleavedcaspase3/9 and caspase3/9 activity ( Figure S3K-N). These results indicate that in response to ox-LDL, IRGM/Irgm1 deficiency decreased macrophage apoptosis in the late-stage in vitro. ...
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Interleukin (IL)-17A plays a role in the development of atherosclerotic plaques; however, the mechanism remains unclear. In this study, apolipoprotein E-deficient (ApoE–/–) mice were fed a high-fat diet to induce atherosclerosis, followed by the treatment with exogenous recombinant IL-17A or the neutralizing antibody to confirm the impact of IL-17A...
Citations
... Apoptosis may facilitate macrophage removal from atherosclerotic plaques. As a result of their ability to consume fats and produce foam cells, macrophages are essential to the development of atherosclerosis [85]. These foam cells may undergo apoptosis and contribute to the necrotic cores of atherosclerotic plaques [86]. ...
... It notes that foam cells undergo various programmed cell death pathways, including apoptosis, contributing to the necrotic cores of atherosclerotic plaques. [85] This paper explores the role of TET2 in vascular smooth muscle cell apoptosis and its implications for transplant vasculopathy. It suggests that TET2 expression is negatively regulated in human coronary allograft vasculopathy and that TET2 depletion results in increased apoptosis and medial thinning. ...
Interferon Gamma Inducible Protein 16 (IFI16) belongs to the HIN-200 protein family and is pivotal in immunological responses. Serving as a DNA sensor, IFI16 identifies viral and aberrant DNA, triggering immune and inflammatory responses. It is implicated in diverse cellular death mechanisms, such as pyroptosis, apoptosis, and necroptosis. Notably, these processes are integral to the emergent concept of PANoptosis, which encompasses cellular demise and inflammatory pathways. Current research implies a significant regulatory role for IFI16 in PANoptosis, particularly regarding cardiac pathologies. This review delves into the complex interplay between IFI16 and PANoptosis in heart diseases, including atherosclerosis, myocardial infarction, heart failure, and diabetic cardiomyopathy. It synthesizes evidence of IFI16’s impact on PANoptosis, with the intention of providing novel insights for therapeutic strategies targeting heart diseases.
... Reactive oxygen species (ROS) are the main type of free radical present in biological systems. ROS are usually produced in mitochondria 8, 9 . Low levels of ROS result in the initiation, development, and progression of malignancies, whereas high levels of ROS can lead to apoptosis of cancer cells 10,11 . ...
Myelodysplastic syndromes (MDS) are clonal hematopoietic malignancies and seriously threatens people’s health. Current therapies include bone marrow transplantation and several hypomethylating agents. However, many elderly patients cannot benefit from bone marrow transplantation and many patients develop drug resistance to hypomethylating agents, making it urgent to explore novel therapy. RSL3 can effectively induce ferroptosis in various tumors and combination of RSL3 and hypomethylating agents is promising to treat many tumors. However, its effect in MDS was unknown. In this study, we found that RSL3 inhibited MDS cell proliferation through inducing ROS-dependent apoptosis. RSL3 inhibited Bcl-2 expression and increased caspase 3 and PARP cleavage. RNA-seq analysis revealed that MYB may be a potential target of RSL3. Rescue experiments showed that overexpression of MYB can rescue MDS cell proliferation inhibition caused by RSL3. Cellular thermal shift assay showed that RSL3 binds to MYB to exert its function. Furthermore, RSL3 inhibited tumor growth and decreased MYB and Bcl-2 expression in vivo. More importantly, RSL3 decreased the viability of bone marrow mononuclear cells (BMMCs) isolated from MDS patients, and RSL3 had a synergistic effect with DAC in MDS cells. Our studies have uncovered RSL3 as a promising compound and MYB/Bcl-2 signaling pathway as a potential target for MDS treatment.
... Cold-induced increases in serum IL-2, IL-6, and TNF-α levels enhance the inflammatory response within the plaque. Cold also activates the pro-inflammatory phenotype of macrophages, leading to increased macrophage accumulation in the plaque, heightening plaque vulnerability and pressure [42][43] . Consequently, this escalation contributes to increased plaque rupture and adverse cardiovascular events, including death, in patients exposed to cold. ...
Cold stimulation has been linked to acute myocardial infarction and other cardiovascular diseases. Residents in the frigid zones, such Heilongjiang Province, experience a higher incidence of adverse cardiovascular events during winter, posing a significant health threat and increasing the overall medical burden. Cold stimulation serves as an detrimental stressor, inducing inflammation in the body. Therefore, understanding the role of inflammatory responses induced by cold stimulation in the occurrence and development of cardiovascular diseases is of paramount importance. Given the impact of cold on inflammation in cardiovascular diseases and the expanding array of anti-inflammatory methods for the treatment of cardiovascular diseases, delving into the inflammatory responses mediated by can significantly complement cardiovascular disease management. This review explorest the synergistic relationship between cold stimulation and inflammation induction, elucidating how this interplay influences the occurrence and progression of cardiovascular diseases.
... 31 MAPK signaling pathway may be involved in mediating neuronal apoptosis in IS. 32 MAPK signaling pathway is also involved in regulating inflammation, oxidative stress, cognitive impairment and neuronal damage in IS. 33 Moreover, MAPK signaling pathway may also be involved in the regulation of atherosclerotic plaque stability. 34 A previous study showed that Ginsenoside Rb1 may improve the functional recovery after stroke by regulating cAMP/PKA/CREB signaling pathway to stimulate axon regeneration and brain repair. 35 CAMP also plays a role in immune regulation of acute IS. 36 In addition, cAMP also plays a role in the formation and progression of atherosclerosis. ...
Background
The specific molecular mechanistic link between atherosclerotic plaques and ischemic stroke (IS) is not clear. The aim of this study is to explore the potential molecular relationship between atherosclerotic plaques and IS.
Methods
All data were downloaded from the Gene Expression Omnibus (GEO) database. Key hub differentially expressed mRNAs (DEmRNAs) related to atherosclerotic plaques and IS were identified by differential expression analysis and least absolute shrinkage and selection operator (LASSO) analysis. Subsequently, a diagnostic model was established based on the expression of key hub DEmRNAs and logistic regression. In order to understand the molecular mechanism of key hub DEmRNAs, the transcription factor (TF) regulatory network and mRNA-miRNA-lncRNA regulatory network were also constructed. In addition, functional enrichment analysis and single-sample Gene Set Enrichment Analysis (ssGSEA) analysis were also performed.
Results
Four key hub DEmRNAs (ADCY3, CLDN7, PPM1B and RRAS2) were identified by differential expression analysis and LASSO analysis. Moreover, the diagnostic model based on four key hub DEmRNAs has excellent diagnostic accuracy. We also found that Type 1 T helper cell may be associated with IS caused by atherosclerosis based on ssGSEA analysis. In the mRNA-miRNA-lncRNA regulatory network, we found that multiple signaling axes such as RRAS2-hsa-miR-3150b-3p-ILF3-AS1, PPM1B-hsa-miR-541-5p-LINC00294, CLDN7-hsa-miR-184-LINC00467 and ADCY3-hsa-miR-488-3p-URB1-AS1 may play an important role in the progression of IS. In addition, some signaling pathways, including chemokine signaling pathway, MAPK signaling pathway and cAMP signaling pathway, may be involved in regulating IS.
Conclusion
The identified key molecules, signaling pathways and immune cells may help to provide a theoretical basis for exploring the relationship between atherosclerotic plaque and the progression of IS.
... The aortic root was embedded in paraffin, and then the aortic root was cut into 5 μm slices from the aortic valve. Three consecutive slices were dewaxed with xylene, then hydrated with alcohol from low to high concentration, and the morphological analysis was carried out by hematoxylin-eosin and Movat five-color dyeing in references [21]. Moreover, Three consecutive slices were fixed, and fluorescent antibody nitric oxide synthase (iNOS) and mannose receptor 1 (CD206) diluted with a certain staining titer were dripped, and the staining analysis was carried out according to the literature to evaluate the polarization level of macrophages in aortic plaque [22]. ...
Corilagin, a polyphenolic tannic acid compound, showed significant anti-inflammatory activity in atherosclerotic mice. The present study aimed to evaluate the effect and mechanism of corilagin in atherosclerosis by in vivo, in vitro and in molecular docking strategies analysis. An atherosclerotic model was established by feeding ApoE−/− mice a high-fat diet. Murine RAW264.7 macrophages were cultured and induced with lipopolysaccharide (LPS). Treatment with corilagin had a marked inhibitory effect on the plaque area and lipid accumulation in atherosclerotic mice. Corilagin decreased the expression of iNOS and promoted the expression of CD206 in aortic plaque, as well as inhibited the production of proinflammatory factors in HFD-fed ApoE−/− mice and LPS-induced RAW264.6 cell. Corilagin also obviously inhibited the expression of TLR4, reduced the phosphorylation of the JNK, the protein expressions of p38 and NF-κB pathway. In addition, corilagin markedly diminished the nuclear translocation of NF-κBp65. Similarly, molecular docking study suggested that hydrogen bonds were detected between the corilagin and the five proteins (TLR4, Myd88, p65, P38, and JNK) with a significant “CDOCKER energy”. These results showed that the antiatherosclerotic effect of corilagin against M1 macrophage polarization and inflammation via suppression the activation of TLR4-NFκB/MAPK signaling pathway. Therefore, corilagin could be a promising lead compound to develop drugs for the treatment of atherosclerosis.
... Histogram showed the result of quantization of green fluorescence intensity in the image by Image J software. On the other hand, ROS accumulation leads to the breakdown of intracellular nucleic acid (DNA) and even apoptosis [27]. TUNEL probe can label terminal transferase and accurately reflect the characteristics of apoptotic cells. ...
Background: Influenza A (H1N1) virus is an acute respiratory infectious disease that causes massive morbidity and mortality worldwide. As an essential trace element, selenium is widely applied in the treatment of various diseases because of its functions of enhancing immune response, antioxidant and antiviral mutation. In this study, we constructed the selenium-containing metal complex drug delivery system Ru(biim)(PhenSe)2 (RuSe), and investigated the anti-influenza virus efficacy and the potential antiviral mechanism for RuSe.
Methods: The inhibitory effect of RuSe on influenza-mediated apoptosis was examined by cell count assay, cell cycle assay, Annenxin-V assay, TUNEL-DAPI assay and reactive oxygen species level determination. Virulence assay, PCR and neuraminidase inhibition assay revealed the inhibition of RuSe on influenza virus. At the level of animal experiments, two animal models were used to clarify the role of RuSe through HE staining, immunohistochemical staining, cytokine determination, selenium metabolism determination and selenium protein expression level determination.
Results: The results of this study confirm that RuSe enhances the expression levels of selenium proteins GPx1 and TrxR1 by regulating selenium metabolism, thereby inhibiting viral replication and assembly and regulating virus-mediated mitochondria-related apoptosis. On the other hand, animal experiments show that RuSe can reduce lung tissue inflammation and inhibit lung tissue cell apoptosis in mice, and improve the survival state of mice. In addition, RuSe significantly improves the low immune response of Se-deficient mice by regulating selenium metabolism, and effectively alleviated lung fibrosis and lung tissue apoptosis in Se-deficient mice.
Conclusions: This study suggests that RuSe provides a promising new approach for the clinical treatment of influenza virus.
... RAS, a member of the GTPase family, is a small monomeric GTP-binding protein composed of 190 amino acid residues. [30][31][32][33] It has GTPase activity and is located on the cytoplasmic side of the plasma membrane. [34][35][36] The RAS protein binds to the N-terminal domain of RAF and activates the RAS/RAF/ERK pathway. ...
The clinical application of anthracyclines such as doxorubicin (DOX) is limited due to their cardiotoxicity. N6-methyladenosine (m6A) plays an essential role in numerous biological processes. However, the roles of m6A and m6A demethylase ALKBH5 in DOX-induced cardiotoxicity (DIC) remain unclear. In this research, DIC models were constructed using Alkbh5-knockout (KO), Alkbh5-knockin (KI), and Alkbh5-myocardial-specific knockout (ALKBH5flox/flox, αMyHC−Cre) mice. Cardiac function and DOX-mediated signal transduction were investigated. As a result, both Alkbh5 whole-body KO and myocardial-specific KO mice had increased mortality, decreased cardiac function, and aggravated DIC injury with severe myocardial mitochondrial damage. Conversely, ALKBH5 overexpression alleviated DOX-mediated mitochondrial injury, increased survival, and improved myocardial function. Mechanistically, ALKBH5 regulated the expression of Rasal3 in an m6A-dependent manner through posttranscriptional mRNA regulation and reduced Rasal3 mRNA stability, thus activating RAS3, inhibiting apoptosis through the RAS/RAF/ERK signaling pathway, and alleviating DIC injury. These findings indicate the potential therapeutic effect of ALKBH5 on DIC.
... A combinational treatment of resveratrol and cisplatin can increase the level of phosphorylated eIF2α, which leads to increased intracellular Ca 2+ levels, thereby triggering endoplasmic reticulum stress and apoptosis of gastric cancer cells [72]. Alongside these treatments, the accumulation of intracellular reactive oxygen species (ROS) has also been shown to be associated with the induction of apoptosis [73]. Agrimoniin is a type of polyphenolic compound derived from Agrimonia pilosa ledeb, a perennial herb that has been widely used in traditional Chinese medicine [74]. ...
Polyphenols and their derivates, a kind of natural product distributed in herb plants, vegetables, and fruits, are the most abundant antioxidants in the human diet and have been found to display cancer-preventative effects in several epidemiological studies. The scientific community has also validated the anti-cancer bioactivities and low toxicities of polyphenolic compounds, including flavones, tannins, phenolic acids, and anthocyanins, through in vitro and in vivo studies. However, the low stability, weak targeting ability, poor solubility, and low bioavailability of pure polyphenolic agents have significantly impaired their treatment efficacy. Nowadays, nano-based technology has been applied to surmount these restrictions and maximize the treatment efficacy of polyphenols. In this review, we summarize the advantages and related mechanisms of polyphenols in cancer treatment. Moreover, aiming at the poor solubility and low bioavailability of pure polyphenols in vivo, the advantages of nano-based delivery systems and recent research developments are highlighted. Herein, particular emphasis is mainly placed on the most widely used nanomaterials in the delivery of natural products, including liposomes, micelles, and nanogels. Finally, we present an overview and the challenges of future implementations of nano-based delivery systems of polyphenolic compounds in the cancer therapeutic field.
... Nowadays, emerging evidence have suggested that apoptosis, associated with ECs, VSMCs, and MDMs [38][39][40], is involved in the progression of AS [41,42], effecting the leukocyte infiltration, fibrous cap thinness, and lipid core expansion [43][44][45]. In addition, increased apoptotic cells have already been observed in atherosclerotic arteries compared with those in normal arteries [46], indicating there might be a tide association between apoptosis and AS. ...
Nowadays, as a type of orderly and active death determined by genes, programmed cell death (PCD), including apoptosis, pyroptosis, ferroptosis, and necroptosis, has attracted much attention owing to its participation in numerous chronic cardiovascular diseases, especially atherosclerosis (AS), a canonical chronic inflammatory disease featured by lipid metabolism disturbance. Abundant researches have reported that PCD under distinct internal conditions fulfills different roles of atherosclerotic pathological processes, including lipid core expansion, leukocyte adhesion, and infiltration. Noteworthy, emerging evidence recently has also suggested that oxidative stress (OS), an imbalance of antioxidants and oxygen free radicals, has the potential to mediate PCD occurrence via multiple ways, including oxidization and deubiquitination. Interestingly, more recently, several studies have proposed that the mediating mechanisms could effect on the atherosclerotic initiation and progression significantly from variable aspects, so it is of great clinical importance to clarify how OS-mediated PCD and AS interact. Herein, with the aim of summarizing potential and sufficient atherosclerotic therapy targets, we seek to provide extensive analysis of the specific regulatory mechanisms of PCD mediated by OS and their multifaceted effects on the entire pathological atherosclerotic progression.
... A recent study suggested that RIOK2 is a new target of the MAPK-activated kinase RSK [39]. As components of the MAPK signaling pathway, ERK and JNK are associated with the regulation of cell survival and death [40,41]. In this study, knockdown of RIOK2 also significantly increased the phosphorylation of ERK and JNK, representing the activation of MAPKs in DON-induced IPEC-J2. ...
Deoxynivalenol (DON) is a type of mycotoxin that threatens human and livestock health. Right open reading frame kinase 2 (RIOK2) is a kinase that has a pivotal function in ribosome maturation and cell cycle progression. This study aims to clarify the role of the RIOK2 gene in DON-induced cytotoxicity regulation in porcine intestinal epithelial cells (IPEC-J2). Cell viability assay and flow cytometry showed that the knockdown of RIOK2 inhibited proliferation and induced apoptosis, cell cycle arrest, and oxidative stress in DON-induced IPEC-J2. Then, transcriptome profiling identified candidate genes and pathways that closely interacted with both DON cytotoxicity regulation and RIOK2 expression. Furthermore, RIOK2 interference promoted the activation of the MAPK signaling pathway by increasing the phosphorylation of ERK and JNK. Additionally, we performed the dual-luciferase reporter and ChIP assays to elucidate that the expression of RIOK2 was influenced by the binding of transcription factor Sp1 with the promoter region. Briefly, the reduced expression of the RIOK2 gene exacerbates the cytotoxic effects induced by DON in IPEC-J2. Our findings provide insights into the control strategies for DON contamination by identifying functional genes and effective molecular markers.
