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ALK5 inhibitor suppressed TGFβ-induced VEGF mRNA expression and VEGF protein production. VEGF mRNA by RT-PCR (a) and VEGF protein concentration by ELISA (b). Synovial fibroblasts were stimulated with α-MEM (control), hrTGFβ (TGFβ), or hrTGFβ + SB505124 (TGFβ + SB505124) for 6 hours prior to RT-PCR or ELISA. Values represent mean ± SE (n = 8). ∗p<0.05 compared to control.
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Background:
Previous studies suggest the presence of an association of vascular endothelial growth factor (VEGF) with osteoarthritis (OA) severity and pain in patients with knee OA. VEGF expression in human synovial fibroblasts (SFs) is induced by transforming growth factor-beta (TGFβ). However, the signaling pathway governing TGFβ-mediated regula...
Citations
... In mouse dendritic cells, TGF-β1 was shown to enhance the expression of vascular endothelial growth factor receptor Flt-1 (VEGFR1) and protein expression through the typical Smad pathway (Nam et al., 2010). Similarly, the canonical and non-canonical TGF-β1 pathway regulates VEGF mRNA and protein expression in synovial fibroblasts derived from osteoarthritis (Takano and Uchida, 2019). ...
Benign prostatic hyperplasia (BPH) is a common male disorder. Febuxostat is a non-purine, selective inhibitor of xanthine oxidase (XO), which has a strong antioxidant capacity and pleiotropic pharmacological properties. This study’s objective was to explore the potential ameliorative effects of febuxostat against testosterone-induced BPH in rats. Febuxostat (10 mg/kg/day, per os [p.o.]) prevented increased prostate index levels, serum levels of prostate-specific antigen (PSA), and testosterone levels compared to animals treated with testosterone alone,
when administered for 28 days. Histological examination indicated that febuxostat dramatically ameliorated pathological changes in the prostate architecture compared to the testosterone group. Similarly, febuxostat markedly improved testosterone-induced oxidative stress by inhibiting the increase in lipid peroxide and nitrite content, and by reducing the level of depletion of reduced glutathione (GSH) and superoxide dismutase (SOD) activity, which significantly reduced the prostate content of pro-inflammatory cytokines, including tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6). Furthermore, febuxostat significantly reduced the prostatic
content, both in terms of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) messenger ribonucleic acid (mRNA) levels, and of protein levels. Moreover, compared to the testosterone group, febuxostat’s
beneficial effects prevented the increase in growth factors, comprising vascular endothelial cell growth factor A (VEGF-A) and transforming growth factor beta (TGF-β) protein levels. Its ameliorating effects were equal to those of finasteride, which is the most widely used remedy for BPH. In conclusion, this study provides novel evidence that febuxostat experimentally attenuates testosterone-induced BPH in rats, at least in part by inhibiting iNOS/COX-2 and VEGF/TGF-β pathways.
... Further, TAK1 has many targets outside of p38 MAPKs which could additionally play a role in aggresome formation such as c-Jun N-terminal kinases (JNKs) and Nuclear Factor kappa-light-chain-enhancer of activated B cells (NF-kB) 40 . Lastly, although TAK-715 and 5Z-7-oxozeaenol are recognized as inhibitors of p38α and p38β MAPKs or TAK1 respectively 21,27,37,41 , both of these drugs are known to have off-target effects. Therefore, it is possible that the effects we observe from these drugs on aggresome formation is through their effect on other proteins. ...
The aggresome is a protein turnover system in which proteins are trafficked along microtubules to the centrosome for degradation. Despite extensive focus on aggresomes in immortalized cell lines, it remains unclear if the aggresome is conserved in all primary cells and all cell-states. Here we examined the aggresome in primary adult mouse dermal fibroblasts shifted into four distinct cell-states. We found that in response to proteasome inhibition, quiescent and immortalized fibroblasts formed aggresomes, whereas proliferating and senescent fibroblasts did not. Using this model, we generated a resource to provide a characterization of the proteostasis networks in which the aggresome is used and transcriptomic features associated with the presence or absence of aggresome formation. Using this resource, we validate a previously reported role for p38 MAPK signaling in aggresome formation and identify TAK1 as a novel driver of aggresome formation upstream of p38 MAPKs. Together, our data demonstrate that the aggresome is a non-universal protein degradation system which can be used cell-state specifically and provide a resource for studying aggresome formation and function.
... In addition, upregulation of pro-inflammatory (IL-7, IL-12, IFNgamma) cytokines also correlated significantly with the level of pain in knee OA patients, and the synovial inflammatory mediators (IL-6, IL-8, IFN-γ, SCGF-β, CXCL1) showed significant associations to OA severity 128 . TGF-β was suggested to regulate VEGF expression in human synovial fibroblasts through both the canonical and noncanonical pathways 130 . Thus, inhibition of TGF-β and VEGF signaling (e.g., VEGFR1 and VEGFR2) may reduce OA pain and progression. ...
The field of research on pain originating from various bone diseases is expanding rapidly, with new mechanisms and targets asserting both peripheral and central sites of action. The scope of research is broadening from bone biology to neuroscience, neuroendocrinology, and immunology. In particular, the roles of primary sensory neurons and non-neuronal cells in the peripheral tissues as important targets for bone pain treatment are under extensive investigation in both pre-clinical and clinical settings. An understanding of the peripheral mechanisms underlying pain conditions associated with various bone diseases will aid in the appropriate application and development of optimal strategies for not only managing bone pain symptoms but also improving bone repairing and remodeling, which potentially cures the underlying etiology for long-term functional recovery. In this review, we focus on advances in important preclinical studies of significant bone pain conditions in the past 5 years that indicated new peripheral neuronal and non-neuronal mechanisms, novel targets for potential clinical interventions, and future directions of research.
... COL10A1 is considered as the standard marker and RUNX2 as the master transcription factor of chondrocyte hypertrophy [2,34,[93][94][95][96][97]. IHH is a key regulator of endochondral ossification and is mainly produced and secreted by pre-hypertrophic chondrocytes [98][99][100], while VEGFA is mainly found in the late hypertrophic phase, correlating with osteoblast formation [95,[101][102][103]. Lastly, TGF-β increased COL1A1 expression, which is associated with dedifferentiation towards fibrotic chondrocytes. ...
During osteoarthritis (OA), hypertrophy-like chondrocytes contribute to the disease process. TGF-β’s signaling pathways can contribute to a hypertrophy(-like) phenotype in chondrocytes, especially at high doses of TGF-β. In this study, we examine which transcription factors (TFs) are activated and involved in TGF-β-dependent induction of a hypertrophy-like phenotype in human OA chondrocytes. We found that TGF-β, at levels found in synovial fluid in OA patients, induces hypertrophic differentiation, as characterized by increased expression of RUNX2, COL10A1, COL1A1, VEGFA and IHH. Using luciferase-based TF activity assays, we observed that the expression of these hypertrophy genes positively correlated to SMAD3:4, STAT3 and AP1 activity. Blocking these TFs using specific inhibitors for ALK-5-induced SMAD signaling (5 µM SB-505124), JAK-STAT signaling (1 µM Tofacitinib) and JNK signaling (10 µM SP-600125) led to the striking observation that only SB-505124 repressed the expression of hypertrophy factors in TGF-β-stimulated chondrocytes. Therefore, we conclude that ALK5 kinase activity is essential for TGF-β-induced expression of crucial hypertrophy factors in chondrocytes.
Aim. We aimed to evaluate levels of transforming growth factor β1 (TGF-β1) in the serum, lymph nodes, and primary tumour in patients with colorectal cancer. Materials and Methods. Here we enrolled 44 patients with colorectal cancer and 25 patients with benign tumours of the colon admitted to Chita Regional Cancer Centre in 2019-2020. The control group included 25 patients with colon injury. The concentration of TGF-β1 in the serum, lymph nodes, and tumour homogenate was measured by flow cytometry (CytoFlex LX analyzer and LEGENDplex HU multiplex analysis kit). Results. Serum level of TGF-β1 in patients with colorectal cancer was 1.58-fold lower than in those with benign colon tumours and 1.38-fold lower than in the control group. In contrast, TGF-β1 level in tumor tissue was 5.91 (3.86; 7.81) fold higher than in the injured colonic tissue from the control group, although there were no statistically significant differences between the cancerous tissue and benign neoplasms. Conclusion. TGF-β1 is increased in tumour tissue but reduced in the serum of patients with colorectal cancer.
Background
There is growing evidence that cytokines and adipokines are associated with osteoarthritis (OA) severity, progression, and severity of associated pain. However, the cytokine response to total knee arthroplasty (TKA) and its association with persistent postoperative pain is not well understood. This study aims to describe the perioperative systemic (plasma) and local (synovial fluid) cytokine profiles of patients who do and do not develop persistent pain after TKA.
Methods
Patients undergoing primary unilateral TKA for end-stage OA were prospectively enrolled. Demographic and clinical data were gathered preoperatively and postoperatively. Synovial fluid was collected pre arthrotomy and plasma was collected at multiple time points before and after surgery. Persistent postoperative pain (PPP) was defined as Numerical Rating Score≥4 at 6 months. Cytokine levels were measured using the V-Plex Human Cytokine 30-Plex Panel (Mesoscale—Rockville, Maryland, USA). Cytokine levels were compared between PPP and minimal pain groups. Given that the study outcomes are exploratory, no adjustment was performed for multiple testing.
Results
Incidence of persistent pain at 6 months post TKA was 15/162 (9.3%). Postoperative plasma levels of four cytokines were significantly different in patients who developed persistent postoperative pain: interleukin (IL)-10, IL-1β, vascular endothelial growth factor, and IL12/IL23p40. Significantly lower IL-10 levels in the prearthrotomy synovial fluid were associated with development of postoperative persistent pain.
Conclusions
This prospective cohort study described a distinct acute perioperative inflammatory response profile in patients who developed persistent post-TKA pain, characterized by significant differences in four cytokines over the first 2 postoperative days. These results support the growing evidence that the patient-specific biologic response to surgery may influence longer-term clinical outcomes after TKA.
Trial registration number
Clinicaltrials.gov NCT02626533 .
The aggresome is a protein turnover system in which proteins are trafficked along microtubules to the centrosome for degradation. Despite extensive focus on aggresomes in immortalized cell lines, it remains unclear if the aggresome is conserved in all primary cells and all cell-states. Here we examined the aggresome in primary adult mouse dermal fibroblasts in four distinct cell-states. We found that in response to proteasome inhibition, quiescent and immortalized fibroblasts formed aggresomes whereas proliferating and senescent fibroblasts did not. Transcriptomic analysis of the fibroblast cell-state-specific response to proteasome inhibition revealed that stress-activated MAPK signaling was associated with aggresome formation. Supporting a functional role for stress-activated MAPK signaling in aggresome formation, inhibition of TAK1 and p38α/β MAPKs suppressed aggresome formation. Together, our data suggest that the aggresome is a non-universal protein degradation system that forms through stress-activated MAPK signaling which can be used cell-state specifically.
Activation of fibroblasts is a key event during normal tissue repair after injury and the dysregulated repair processes that result in organ fibrosis. To most researchers, fibroblasts are rather unremarkable spindle‐shaped cells embedded in the fibrous collagen matrix of connective tissues and/or deemed useful to perform mechanistic studies with adherent cells in culture. For more than a century, fibroblasts escaped thorough classification due to the lack of specific markers and were treated as the leftovers after all other cells have been identified from a tissue sample. With novel cell lineage tracing and single cell transcriptomics tools, bona fide fibroblasts emerge as only one heterogeneous sub‐population of a much larger group of partly overlapping cell types, including mesenchymal stromal cells, fibro‐adipogenic progenitor cells, pericytes, and/or perivascular cells. All these cells are activated to contribute to tissue repair after injury and/or chronic inflammation. “Activation” can entail various functions, such as enhanced proliferation, migration, instruction of inflammatory cells, secretion of extracellular matrix proteins and organizing enzymes, and acquisition of a contractile myofibroblast phenotype. We provide our view on the fibroblastic cell types and activation states playing a role during physiological and pathological repair and their crosstalk with inflammatory macrophages. Inflammation and fibrosis of the articular synovium during rheumatoid arthritis and osteoarthritis are used as specific examples to discuss inflammatory fibroblast phenotypes. Ultimately, delineating the precursors and functional roles of activated fibroblastic cells will contribute to better and more specific intervention strategies to treat fibroproliferative and fibrocontractive disorders.
Dermal papilla cells (DPCs), an important component of hair follicles, its proliferation and apoptosis directly regulate and maintain the growth of hair follicles. All-trans-retinoic acid (ATRA) plays a critical role in hair growth. In this study, the effects of ATRA on cultured mink hair follicle growth were studied by administration of different concentrations of ATRA for 12 days in vitro. In addition, the proliferation and apoptosis of DPCs were measured after treating with ATRA. The mRNA and protein levels of hair follicle growth associated factor transforming growth factor-β2 (TGF-β2) and the phosphorylation levels of Smad2/3 were determined. Moreover, TGF-β type I and type II receptor inhibitor LY2109761 and specific inhibitor of Smad3 (SIS3) were administered to investigate the underlying molecular mechanism. The results showed that ATRA inhibited hair follicle growth, promoted TGF-β2 expression and activated phosphorylation of Smad2/3. In addition, ATRA inhibited cell proliferation by arresting the cell cycle at G1 phase and induced apoptosis of DPCs by enhancing the ratio of Bax/Bcl-2 and promoted the cleavage of caspase-3. Furthermore, LY2109761 or SIS3 partially reversed the decreased cell viability, increased apoptosis that were induced by ATRA. In conclusion, ATRA could inhibit hair follicle growth via inhibiting proliferation and inducing apoptosis of DPCs partially through the TGF-β2/Smad2/3 pathway.
