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CTRP3 Decreases Inflammatory Factors Produced by LPS-triggered Foam Cells. THP-1 cells and mouse peritoneal macrophages (1 × 10 6 cells/mL/well) were preincubated with different concentrations of CTRP3 (0-1.0 μg/ mL) for 30 min. After LPS and ox-LDL stimulation, the supernatant protein levels of TNFα, IL-6, MCP-1, IL-1β and MMP-9 were evaluated by ELISA. At least three independent experiments were performed. (mean ± SD, * : compared to the control group, P < 0.05, # : compared to the LPS and ox-LDL treatment group, P < 0.05).
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Macrophage inflammation and foam cell formation are critical events during the initiation and development of atherosclerosis (AS). C1q/tumor necrosis factor-related protein-3 (CTRP3) is a novel adipokine with anti-inflammatory and cardioprotection properties; however, little is known regarding the influence of CTRP3 on AS. As macrophages play a key...
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Context 1
... inflammatory reaction induced by TLR-4 ligands (such as LPS) is an important event in AS, and studies have identified CTRP3 as a negative regulator of inflammatory responses in monocytes and 3T3-L1 adipocytes stimulated by LPS [13,14]. To explore whether CTRP3 has an anti-inflammatory function in LPS- triggered foam cells, THP-1 cells and mouse peritoneal macrophages were cultured as mentioned above, and the effects of CTRP3 on LPS-induced release of TNF-α, IL-6, MCP-1, IL-1β and MMP-9 were investigated by ELISA. As shown in Figure 2, LPS and ox-LDL stimulation prominently elevated supernatant TNF-α, IL-6, MCP-1, IL-1β and MMP-9 levels in both THP-1 macrophages (A) Impacts of increasing CTRP3 concentration on the viability of THP-1 differentiated macrophages and mouse peritoneal macrophages. Cells were suspended into 1 × 10 4 cells /100μL/well; THP-1 cells were induced to differentiate into macrophages by culture with 100 ng/mL PMA for 48 h. Mouse peritoneal macrophages were extracted and cultured as mentioned above, after which increasing doses of CTRP3 (0-10 μg/ mL) were added; 24 h later, the CCK-8 assay was used to detect viability (mean ± SD). (B-C) Impacts of CTRP3 on LPS-triggered macrophage lipid deposition and foam cell formation in both kinds of macrophages (1 × 10 6 cells/mL/well). The cholesterol content was detected using a cholesterol quantification kit to evaluate lipid deposition in LPS-triggered macrophages (B, mean ± SD, * : Compared to the control group, P < 0.05; # : compared to the ox-LDL and LPS treatment group, P < 0.05). Oil red O staining was performed to analyze the effect of CTRP3 on foam cell formation (C, THP-1 cells: 200×, mouse peritoneal macrophages: 400×). At least three independent experiments were performed. and mouse peritoneal macrophages (P < 0.001), and the pro-inflammatory impact of LPS and ox-LDL was dose- dependently antagonized by CTRP3 at 0.125 -1 μg/mL (P < 0.05). Moreover, a concentration of 1 μg/mL CTRP3 was effective for all five inflammatory cytokines in both cell lines. As 1 μg/mL CTRP3 significantly regulated lipid metabolism in foam cells, we chose this concentration for further experiments. According to the CCK-8 assay, CTRP3 has no cytotoxic effect on macrophages; therefore, we excluded a cytotoxic effect of CTRP3 as responsible for the observed reduction in cytokine ...
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Citations
... Megf6 is located in a collagen-containing extracellular matrix and regulates zebrafish's cartilage and bone formation [28]. C1qtnf3 relieves the inflammation induced by LPS through the PPARγ/TLR4 pathway [29] and improves the proliferation of chondrogenic precursors and chondrocytes during chondrogenic differentiation [30]. The hypomethylation of Isoc2b is associated with age in mice [31], but studies on its function are still lacking. ...
Background: Osteoarthritis (OA) is the most common age-related joint disease, and the NLRP3-induced pyroptosis has been demonstrated in its progression. The upstream molecules or specific mechanisms controlling NLRP3 and pyroptosis in OA remain unclear.
Methods: Transcriptome sequencing was performed in the OA mice model, and the expression levels of differentially expressed genes were assessed by qRT-PCR. The cell model was constructed by IL-1β-induced ATDC5 cells. The cell proliferation was examined using CCK-8 assay, and apoptosis was tested using flow cytometry. Western blot was used in protein inspection, and ELISA was used in inflammatory response evaluation.
Results: Compared with the control group, there were 229 up-regulated and 32 down-regulated genes in model group. We detected that FOXQ1 was down-regulated in the OA mice model, improved proliferation, and restrained apoptosis of chondrocytes. Over-expression of FOXQ1 could inhibit pyroptosis-related proteins and inflammatory cytokines, containing NLRP3, Caspase-1, GSDMD, IL-6, IL-18, and TNF-α, and in contrast, FOXQ1 silencing exerted the opposite trend.
Conclusions: FOXQ1 may inhibit OA progression via down-regulating NLRP3-induced pyroptosis in the present study.
... C1QTNF3 plays an important regulatory role in cartilage development, inflammatory response, cardiovascular disease, adipocyte differentiation, and metabolism (21-24). C1QTNF3 inhibits LPS-induced inflammation via PPAR-g and TLR4-mediated pathways (25). ASFV infection also induces the expression of inflammatory cytokines through the TLRs/MyD88 pathway (26). ...
African swine fever (ASF) is the most dangerous pig disease, and causes enormous economic losses in the global pig industry. However, the mechanisms of ASF virus (ASFV) infection remains largely unclear. Hence, this study investigated the host response mechanisms to ASFV infection. We analyzed the differentially expressed proteins (DEPs) between serum samples from ASFV-infected and uninfected pigs using quantitative proteomics. Setting the p-value < 0.05 and |log2 (fold change)| > 1.5, we identified 173 DEPs, comprising 57 upregulated and 116 downregulated proteins, which belonged to various biological processes and pathways based on the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses. The enriched pathways include immune responses, metabolism, and inflammation signaling pathways. Western blot analysis validated the DEPs identified using quantitative proteomics. Furthermore, our proteomics data showed that C1QTNF3 regulated the inflammatory signaling pathway. C1QTNF3 knockdown led to the upregulation of pro-inflammatory factors IL-1β, IL-8, and IL-6, thus inhibiting ASFV replication. These results indicated that C1QTNF3 was critical for ASFV infection. In conclusion, this study revealed the molecular mechanisms underlying the host-ASFV interaction, which may contribute to the development of novel antiviral strategies against ASFV infection in the future.
... Since its discovery in 2001 (20), a number of experimental in vitro studies have been conducted and C1QTNF3 has been suggested to e.g. stimulate proliferation (21), inhibit LPS-induced inflammatory responses in fibroblasts, adipocytes and macrophages (22)(23)(24)(25) and increase the secretion of adiponectin from adipocytes (21). Moreover, C1QTNF3 has been shown to exert beneficial effects on metabolism and inflammation in vivo; administration or transgenic overexpression of C1QTNF3 attenuated dietinduced hepatic steatosis and lowered glucose levels in Ob/Ob mice (26,27) and C1QTNF3 knockout mice are more susceptible to collagen-induced arthritis in mice (28). ...
... Thus, C1QTNF3 may both enhance and reduce pro-inflammatory responses dependent on the immunological setting. Nevertheless, our results may be particularly difficult to reconcile with a study by Lin and colleagues where wheat germ-produced C1QTNF3 dosedependently inhibited TNF-a, IL-6, MCP1, MMP9 and IL-1b release in LPS-stimulated THP-1 macrophages and mouse peritoneal macrophages (24). In our hands, we saw only small effects of C1QTNF3 treatment on LPS/IFNg-stimulated bone marrow derived macrophages, while a M1-like phenotype was induced in IL-4-stimulated bone marrow derived macrophages. ...
The adipose tissue undergoes substantial tissue remodeling during weight gain-induced expansion as well as in response to the mechanical and immunological stresses from a growing tumor. We identified the C1q/TNF-related protein family member C1qtnf3 as one of the most upregulated genes that encode secreted proteins in tumor-associated inguinal adipose tissue - especially in high fat diet-induced obese mice that displayed 3-fold larger tumors than their lean controls. Interestingly, inguinal adipose tissue C1qtnf3 was co-regulated with several macrophage markers and chemokines and was primarily expressed in fibroblasts while only low levels were detected in adipocytes and macrophages. Administration of C1QTNF3 neutralizing antibodies inhibited macrophage accumulation in tumor-associated inguinal adipose tissue while tumor growth was unaffected. In line with this finding, C1QTNF3 exerted chemotactic actions on both M1- and M2-polarized macrophages in vitro. Moreover, C1QTNF3 treatment of M2-type macrophages stimulated the ERK and Akt pathway associated with increased M1-like polarization as judged by increased expression of M1-macrophage markers, increased production of nitric oxide, reduced oxygen consumption and increased glycolysis. Based on these results, we propose that macrophages are recruited to adipose tissue sites with increased C1QTNF3 production. However, the impact of the immunomodulatory effects of C1QTNF3 in adipose tissue remodeling warrants future investigations.
... CTRP3 is implicated in the development of myocardiac dysfunction, inflammatory bowel diseases, severe acute pancreatitis and chronic kidney diseases (22)(23)(24)(25). CTRP3 also functions as an antagonist for LPS, and modulates antiinflammatory functions of monocytes, macrophages, adipocytes and fibroblasts (26)(27)(28)(29)(30)(31). We identified AdipoR2, but not AdipoR1 nor AdipoR3, as a functional CTRP3 receptor and showed that CTRP3 regulates chondrocyte proliferation via this receptor (32). ...
C1q/TNF-related proteins (CTRP) including CTRP3 are a group of secreted proteins which have a complement C1q-like domain in common, and play versatile roles in lipid metabolism, inflammation, tumor metastasis and bone metabolism. Previously, we showed that the expression of C1qtnf3, encoding CTRP3, is highly augmented in joints of autoimmune arthritis models and CTRP3-deficiency exacerbates collagen-induced arthritis in mice. However, the mechanisms how CTRP3-deficiency exacerbates arthritis still remain to be elucidated. In this study, we showed that CTRP3 was highly expressed in Th17 cell, a key player for the development of autoimmune diseases, and Th17 cell differentiation was augmented in C1qtnf3–/– mice. Th17 cell differentiation, but not Th1 cell differentiation, was suppressed by CTRP3 and this suppression was abolished by the treatment with a receptor antagonist against AdipoR2, but not AdipoR1, associated with suppression of Rorc and Stat3 expression. Furthermore, AdipoR1 and AdipoR2 agonist, AdipoRon suppressed Th17 cell differentiation via AdipoR2, but not AdipoR1. The development of myelin oligodendrocyte glycoprotein (MOG)-induced experimental autoimmune encephalomyelitis was enhanced in C1qtnf3 –/– mice associated with increase of Th17 cell population. CTRP3 inhibited MOG-induced IL-17 production from T cells by affecting both T cells and dendritic cells. These results show that CTRP3 is an endogenous regulator of Th17 differentiation, suggesting that the CTRP3-AdipoR2 axis is a good target for the treatment of Th17 cell-mediated diseases.
... IGF1R-deficient macrophages inhibit the expression of ABCA1 and ABCG1 and reduce lipid efflux and plaque vulnerability [58]. Interestingly, the activation of MC1-R can not only prevent macrophages from accumulating lipid but also promote the reverse transport of cholesterol by up-regulating the levels of ABCA1 and ABCG1 and reducing the expression of CD36 on the cell surface [59,60]. Li et al. found that overexpression of C1q/TNF-related protein 9 (CTRP9) can reduce the levels of pro-inflammatory factors such as TNF-α and MCP-1 in macrophages [61]. ...
Cardiovascular diseases (CVDs) caused by arteriosclerosis are the leading cause of death and disability worldwide. In the late stages of atherosclerosis, the atherosclerotic plaque gradually expands in the blood vessels, resulting in vascular stenosis. When the unstable plaque ruptures and falls off, it blocks the vessel causing vascular thrombosis, leading to strokes, myocardial infarctions, and a series of other serious diseases that endanger people's lives. Therefore, regulating plaque stability is the main means used to address the high mortality associated with CVDs. The progression of the atherosclerotic plaque is a complex integration of vascular cell apoptosis, lipid metabolism disorders, inflammatory cell infiltration, vascular smooth muscle cell migration, and neovascular infiltration. More recently, emerging evidence has demonstrated that non-coding RNAs (ncRNAs) play a significant role in regulating the pathophysiological process of atherosclerotic plaque formation by affecting the biological functions of the vasculature and its associated cells. The purpose of this paper is to comprehensively review the regulatory mechanisms involved in the susceptibility of atherosclerotic plaque rupture, discuss the limitations of current approaches to treat plaque instability, and highlight the potential clinical value of ncRNAs as novel diagnostic biomarkers and potential therapeutic strategies to improve plaque stability and reduce the risk of major cardiovascular events.
... In the present study, we have evaluated the impacts of two adipokines (CTRP15 and adiponectin) on cytokines' expressions and secretions from macrophages. Previous studies demonstrated the CTRP3 and CTRP9 inhibited inflammatory cytokines secretion from human LPS-induced monocytes [19,20]. However, the results of those reports on the impact of adiponectin on inflammation are inconsistent. ...
Atherosclerosis is a progressive inflammatory disease characterized by the accumulation of lipids in the arterial wall. Inflammation plays a key role in the pathogenesis of atherosclerosis and some previous studies have shown the role of adipokines during the inflammatory process of atherosclerosis. Therefore, the present study aimed to evaluate the impacts of adiponectin and CTRP15 on inflammatory cytokines secretions from THP1 and primary macrophages.
Methods:
THP1 monocytes were differentiated to macrophages and primary monocytes were then isolated from patients with coronary artery disease and controls who were differentiated to macrophages. Macrophages were treated with LPS, LPS+adiponectin, and LPS+CTRP15.
Results:
Adiponectin and CTRP15 have reduced IL-6 and TNF-α secretions from LPS-induced THP1 macrophages, and the CTRP15 indicated a more potent anti-inflammatory property compared to adiponectin. In addition, adiponectin reduced cytokines' expressions and secretions in primary macrophages of both patient and control groups. However, CTRP15 has only reduced cytokines' expressions and secretions in controls and it was not able to ameliorate inflammation in macrophages of CAD patients.
Conclusion:
The results of the present study indicate anti-inflammatory impact of adiponectin and CTRP15, while this property was stronger for CTRP15. In addition, it seems likely that anti-inflammatory CTRP15's impact on macrophages in the CAD patients was weaker than macrophages from the controls.
... LPS is known to cause cellular injury that leads to an inflammatory response characterized by production of proinflammatory mediators and accompanied by suppression of macrophage cholesterol efflux genes, including ABCG1 (Shimabukuro Okuda et al., 2020;Park et al., 2012). LPS stimulation induces the classically activated M1 macrophage phenotype, considered to be pro-atherogenic (Lin et al., 2017;Tabas and Bornfeldt, 2016). The finding that OT can modulate cholesterol transport on a cellular transporter level, namely through upregulation of the cholesterol transporter ABCG1 on the macrophage cell membrane, suggests a protective role for OT in inflammatory states that promote lipid accumulation such as atherosclerosis. ...
Introduction and aims
Oxytocin (OT) is a neuropeptide hormone secreted by the posterior pituitary gland. Deficits in OT action have been observed in patients with behavioral and mood disorders, some of which correlate with an increased risk of cardiovascular disease (CVD). Recent research has revealed a wider systemic role that OT plays in inflammatory modulation and development of atherosclerotic plaques. This study investigated the role that OT plays in cholesterol transport and foam cell formation in LPS-stimulated THP-1 human macrophages.
Methods
THP-1 differentiated macrophages were treated with media, LPS (100 ng/ml), LPS + OT (10 pM), or LPS + OT (100 pM). Changes in gene expression and protein levels of cholesterol transporters were analyzed by real time quantitative PCR (RT-qPCR) and Western blot, while ox-LDL uptake and cholesterol efflux capacity were evaluated with fluorometric assays.
Results
RT-qPCR analysis revealed a significant increase in ABCG1 gene expression upon OT + LPS treatment, compared to LPS alone (p = 0.0081), with Western blotting supporting the increase in expression of the ABCG1 protein. Analysis of ox-LDL uptake showed a significantly lower fluorescent value in LPS + OT (100pM) -treated cells when compared to LPS alone (p < 0.0001). While not statistically significant (p = 0.06), cholesterol efflux capacity increased with LPS + OT treatment.
Conclusion
We demonstrate here that OT can attenuate LPS-mediated lipid accumulation in THP-1 macrophages. These findings support the hypothesis that OT could be used to reduce pro-inflammatory and potentially atherogenic changes observed in patients with heightened CVD risk. This study suggests further exploration of OT effects on monocyte and macrophage cholesterol handling in vivo.
... The production and expression of proinflammatory molecules and chemokines have been shown to be increased in obesity (32). The anti-inflammatory role of CTRP3, through suppressing the toll-like receptor 4 (TLR4) signaling pathway (33) and inhibiting TNF-α and IL-6 production in immune cells (34), has been well documented. In agreement with ample evidence for up-regulation of proinflammatory cytokines in the context of obesity in humans (35), we found greater IL-1β, IL-6, MCP-1, and TNF-α expression in adipose tissues from obese patients than from controls. ...
Background:
Obesity, a medical condition with impaired adipokine secretion and function, has a detrimental effect on insulin and glucose metabolism. CTRP3 and CTRP9 are adipokines with possible roles in energy homeostasis regulation. We sought to compare CTRP3, CTRP9, and inflammatory gene expression in subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT) from obese women who underwent bariatric surgery and non-obese women as controls.
Methods:
For this study, the investigators recruited 20 morbidly obese women (BMI> 35) who qualified for bariatric surgery and 20 normal-weight women (BMI< 25) who underwent elective surgeries. Real-time PCR was performed to investigate mRNA expression of CTRP3, CTRP9, and the inflammatory genes IL1-β, IL-6, MCP-1, and TNF-α in SAT and VAT from both obese patients and controls.
Results:
We observed that CTRP3 mRNA levels were significantly greater in VAT from obese patients than from controls (P< 0.0003). Also, patient group had higher levels of CTRP9 that control group (P< 0.0026). Inflammatory cytokines were markedly increased in SAT of obese patients compared to controls (P< 0.05). In addition, our results revealed a positive correlation of CTRP9 with HOMA-IR and waist circumference in VAT and CTRP3 with IL-1β, MCP-1, and TNF-α in SAT.
Conclusion:
Both CTRP3 and CTRP9 expression were significantly higher in VAT from obese patients than from controls, and CTRP3 expression positively correlated with inflammatory parameters. Our findings indicate that CTRP3 and CTRP9 might be important in regulating glucose metabolism and obesity-related conditions such as inflammation.
... [8][9][10] It has also been reported that CTRP3 treatment can reduce the expression of pro-inflammatory cytokines such as TNFα and IL-6 in various cell lines, and plasma levels of CTRP3 decrease in chronic inflammatory diseases. 11,12 Also, a study by Hofmann et al. demonstrated the expression of CTRP3 in mesenteric adipose tissue of the IBD patients; and moreover, they showed that CTRP3 could be antagonizing LPS pathway and exert anti-inflammatory and antifibrotic effects on human primary colonic lamina propria fibroblasts (CLPFs) derivated from the CD patients. 13 However, circulating levels of CTRP3 have not been evaluated in the patients with IBD, and regarding the role of this adipokine in inflammation, the present study evaluated the serum levels of CTRP3 in the patients with UC and CD compared with the control and its relationship with metabolic profile and inflammatory cytokines. ...
Ulcerative colitis (UC) and Crohn's disease (CD) are two major forms of inflammatory bowel disease (IBD), which is an inflammatory disease. Studies have shown that adipose tissue and inflammation play important roles in the pathogenesis of IBD. C1q/TNF‐related protein‐3 (CTRP3) is a newly discovered adipokine playing a substantial role during inflammatory process, and for the first time in the present study, serum levels of this adipokine were measured in the UC and CD patients. This case–control study included 70 control, 50 UC, and 50 CD patients who were diagnosed by standard criteria. Serum levels of adiponectin, IL‐6, TNF‐α, TGF‐β, and CTRP3 were evaluated using ELISA kits. Serum levels of IL‐6, TNF‐α, and TGF‐β elevated in the UC and CD patients compared with the controls while adiponectin and CTRP3 diminished in the patient's groups compared with the control. Furthermore, decrease in CTRP3 serum levels was associated with the risk of UC and CD diseases. Moreover, CTRP3 indicated negative correlation with BMI, FBS, insulin, homeostasis model assessment of insulin resistance, IL‐6, TNF‐α, and TGF‐β and also a positive correlation with adiponectin in both the UC and CD patients. For the first time, the present study demonstrated lower levels of CTRP3 in the UC and CD patients. Decreased serum levels of CTRP3 and its inverse relationship with inflammatory cytokines and TGF‐β levels suggested a possible role for CTRP3 in the pathogenesis of UC and CD diseases.
... In future studies, it will be important to assess JOURNAL OF ORTHOPAEDIC RESEARCH ® MAY 2020 This study was unable to determine whether the reduction in osteoclast number in the KO group is a hematopoietic phenotype in its own right, or is caused by failure of the overall callus to progress into the remodeling stage; if bone fails to form in the central part of the callus, then osteoclast recruitment would be unnecessary. CTRP3 regulates macrophage phenotype commitment and transformation, 24 and future investigation of osteoclastogenesis in the context of CTRP3 deficiency would aid in the interpretation of this study. It is also possible that modification of the initial inflammatory milieu in CTRP3 KO mice could contribute to delay of the entire healing trajectory, though the lack of a phenotype in the rhCTRP3 delivery experiment suggests immunomodulation may not be the primary cause of the fracture healing phenotype. ...
C1q/TNF-related protein 3 (CTRP3) is a cytokine known to regulate a variety of metabolic processes. Though previously undescribed in the context of bone regeneration, high throughput gene expression experiments in mice identified CTRP3 as one of the most highly upregulated genes in fracture callus tissue. Hypothesizing a positive regulatory role for CTRP3 in bone regeneration, we phenotyped skeletal development and fracture healing in CTRP3 knockout (KO) and CTRP3 overexpressing transgenic (TG) mice relative to wild-type (WT) control animals. CTRP3 KO mice experienced delayed endochondral fracture healing, resulting in abnormal mineral distribution, the presence of periosteal marrow compartments, and a nonunion-like state. Decreased osteoclast number was also observed in CTRP3 KO mice, whereas CTRP3 TG mice underwent accelerated callus remodeling. Gene expression profiling revealed a broad impact on osteoblast/osteoclast lineage commitment and metabolism, including arrested progression toward mature skeletal lineages in the KO group. A single systemic injection of CTRP3 protein at time of fracture was insufficient to phenocopy the chronic TG healing response in WT mice. By associating CTRP3 levels with fracture healing progression, these data identify a novel protein family with potential therapeutic and diagnostic value. This article is protected by copyright. All rights reserved.
