TheBDKRB2 +9/-9 Polymorphisms Influence Pro-Inflammatory Cytokine Levels in Knee Osteoarthritis by Altering TLR-2 Expression: Clinical and in Vitro Studies

Background/Aims: The bradykinin B2 receptor (BDKRB2) +9/-9 gene polymorphisms have been shown to be associated with the susceptibility and severity of osteoarthritis (OA); however, the underlying mechanisms are unclear. In this study, we investigated the correlation between the BDKRB2 +9/-9 polymorphisms and pro-inflammatory cytokine levels in OA and the molecular mechanisms involved. Methods: A total of 156 patients with primary knee OA and 121 healthy controls were enrolled. The BDKRB2 +9/-9 polymorphisms were genotyped. The tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and IL-8 levels were determined using Enzyme-linked immunosorbent assay (ELISA). The toll-like receptor (TLR)-2 and TLR-4 mRNA levels were determined by quantitative real-time PCR. The basal and bradykinin-stimulated pro-inflammatory cytokine secretion in human OA synoviocytes and the involvement of TLR-2 and mitogen-activated protein kinases (MAPKs) were investigated. Results: The presence of -9 bp genotype is associated with higher TNF-α, IL-6, and IL-8 levels and higher TLR-2 expression in OA patients. The basal and bradykinin-induced TLR-2 expressions in human OA synoviocytes were significantly reduced by specific inhibitors of p38, JNK1/2, and ERK1/2. Both the B2 receptor antagonist MEN16132 and TLR-2 silencing inhibited IL-6 and IL-8 secretion in human OA synoviocytes. Conclusion: The data suggested that the BDKRB2 +9/-9 polymorphisms influence pro-inflammatory cytokine levels in knee osteoarthritis by altering TLR-2 expression.

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Protective effects of 4, 5-O-dicaffeoylquinic acid against mouse sepsis via down-regulation of TNF-α, IL-6 and IL-8

Tropical Journal of Pharmaceutical Research ◽

10.4314/tjpr.v19i4.6 ◽

2020 ◽

Vol 19 (4) ◽

pp. 715-720

Author(s):

XiaoBo Wang ◽

JianHua Wu ◽

BuKao Ni

Keyword(s):

Survival Rate ◽

Mrna Expression ◽

Molecular Mechanisms ◽

Cecal Ligation ◽

Enzyme Linked Immunosorbent Assay ◽

Protective Effects ◽

Dicaffeoylquinic Acid ◽

Xanthium Sibiricum ◽

Tnf Α

Purpose: To investigate the protective effects of 4, 5-O-dicaffeoylquinic acid (DCQA) isolated from Xanthium sibiricum Patr. against mouse sepsis caused by cecal ligation/puncture (CLP) in vivo, as well as the molecular mechanisms of action involved.Methods: DCQA (7.5, 15, and 30 mg/kg/day) were administered to the mice with sepsis and the survival rate was obtained. Tumor necrosis factor (TNF)-α and interleukin (IL)-6 were examined by enzyme-linked immunosorbent assay (ELISA). Subsequently, the lipopolysaccharide (LPS) neutralizing ability of DCQA (2, 4, and 8 μg/mL) was measured using limulus amebocyte lysate (LAL) test in vitro. Furthermore, the effect of DCQA (10, 20, and 40 μg/mL) on mRNA expression of TNF-α, IL-6, and IL-8 in LPS (100 ng/mL)-treated RAW 264.7 cells was assessed using quantitative real time-polymerase chain reaction (RT-qPCR) assay.Results: DCQA significantly improved the survival rate of mice with sepsis caused by CLP (35, 50, and 65 %, respectively vs. 15 % for control, p < 0.05). LPS levels fell on co-incubation with DCQA in vitro. Moreover, ELISA and RT-qPCR results revealed that DCQA treatment lowered tendency in the mRNA expression of TNF-α, IL-6, and IL-8 (p < 0.01).Conclusion: DCQA exhibits protective effects against sepsis in mice mediated by downregulating TNF-α, IL-6, and IL-8. Further studies, in animals and humans are requied to determine the safety and efficacy of DCQA in both animal and clinical management of sepsis. Keywords: Xanthium sibiricum, 4,5-O-Dicaffeoylquinic acid, Sepsis, Cecal ligation and puncture, Lipopolysaccharide, TNF-α, IL-6, IL-8

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Influence of Isometric Exercise Combined With Electromyostimulation on Inflammatory Cytokine Levels, Muscle Strength, and Knee Joint Function in Elderly Women With Early Knee Osteoarthritis

Frontiers in Physiology ◽

10.3389/fphys.2021.688260 ◽

2021 ◽

Vol 12 ◽

Author(s):

Sunhee Park ◽

Sukyung Min ◽

Si-Hwa Park ◽

Jaehyun Yoo ◽

Yong-Seok Jee

Keyword(s):

Muscle Strength ◽

Knee Osteoarthritis ◽

Inflammatory Cytokine ◽

Elderly Women ◽

Isometric Exercise ◽

Knee Function ◽

Whole Body ◽

Controlled Study ◽

Knee Oa ◽

Cytokine Levels

Background and Objectives: Muscle strengthening exercise is suggested to beneficial for patients with knee osteoarthritis (OA) and electrical muscular stimulation is reported to be effective in improvement of muscle strength. This study examined whether isometric exercise combined with whole body-electromyostimulation (WB-EMS) can improve serum cytokine levels, muscle strength, and knee function in elderly women with early knee OA.Materials and Methods: This randomized controlled study included 75 participants assigned into three groups: the control group (CON), isometric exercise group (ISOM), and isometric exercise and electromyostimulation group (ISOM + EMS). The two exercise groups performed their respective programs for 8 weeks, 3 days a week, 30 min a day. The main exercises for both groups were performed continuously during the 20 min in an alternation of a 6-s contraction with a 4-s break. At pre- and post-intervention, anthropometric variables, muscle strength, Knee Injury and Osteoarthritis Outcome Score (KOOS), and blood sampling for biomarkers including interleukin-6, tumor necrosis factor-α, C-reactive protein, and resistin were performed.Results: All variables at pre-intervention showed no significant differences among the three groups. However, there were significant differences between groups for body composition, muscle strength, KOOS subscale scores, and biomarkers. ISOM + EMS group resulted in a significant reduction in body weight, fat mass, fat percentage, inflammatory cytokine levels, and increased muscle strength. An ISOM + EMS group had the best KOOS score among all groups.Conclusion: Isometric exercise combined with WB-EMS resulted in the best overall improvements in knee function and alleviating the pain and symptoms of patients with early knee OA. Further, reduced levels of inflammatory cytokines were observed. These non-pharmacologic, non-invasive interventions should be considered by healthcare specialists for elderly patients with early knee OA.

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Increased Production of Proinflammatory Cytokines following Infection with Porcine Reproductive and Respiratory Syndrome Virus and Mycoplasma hyopneumoniae

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.11.5.901-908.2004 ◽

2004 ◽

Vol 11 (5) ◽

pp. 901-908 ◽

Cited By ~ 106

Author(s):

Roongroje Thanawongnuwech ◽

Brad Thacker ◽

Patrick Halbur ◽

Eileen L. Thacker

Keyword(s):

Proinflammatory Cytokines ◽

Mycoplasma Hyopneumoniae ◽

Tracheal Ring ◽

In Vivo Model ◽

Respiratory Syndrome Virus ◽

Enzyme Linked Immunosorbent Assay ◽

Mrna Levels ◽

Tnf Α

ABSTRACT Induction of the proinflammatory cytokines interleukin-1 (IL-1) (α and β), IL-6, IL-8, IL-10, IL-12, and tumor necrosis factor alpha (TNF-α) in pulmonary alveolar macrophages (PAMs) was assessed following experimental infection with porcine reproductive and respiratory syndrome virus (PRRSV) and/or Mycoplasma hyopneumoniae by using in vivo and in vitro models. The in vivo model consisted of pigs infected with PRRSV and/or M. hyopneumoniae and necropsied at 10, 28, or 42 days postinfection. Pigs infected with both pathogens had a greater percentage of macroscopic lung lesions, increased clinical disease, and slower viral clearance than pigs infected with either pathogen alone. The pigs infected with both PRRSV and M. hyopneumoniae had significantly increased levels of mRNA for many proinflammatory cytokines in PAMs collected by bronchoalveolar lavage (BAL) at all necropsy dates compared to those in uninfected control pigs. Increased levels of IL-1β, IL-8, IL-10, and TNF-α proteins in BAL fluid, as measured by enzyme-linked immunosorbent assay, confirmed the increased cytokine induction induced by the pathogens. An in vitro model consisted of M. hyopneumoniae-inoculated tracheal ring explants cultured with PRRSV-infected PAMs. PAMs were harvested at 6 or 15 h postinfection with either or both pathogens. The in vitro study detected increased IL-10 and IL-12 mRNA levels in PAMs infected with PRRSV at all time periods. In addition, IL-10 protein levels were significantly elevated in the culture supernatants in the presence of M. hyopneumoniae-inoculated tracheal ring explants. The increased production of proinflammatory cytokines in vivo and in vitro associated with concurrent M. hyopneumoniae and PRRSV infection may play a role in the increased rates of pneumonia associated with PRRSV infection. The increased levels of IL-10 may be a possible mechanism that PRRSV and M. hyopneumoniae use to exacerbate the severity and duration of pneumonia induced by PRRSV and modulate the respiratory immune response.

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Antioxidant glutathione inhibits inflammation in synovial fibroblasts via PTEN/PI3K/AKT pathway: An in vitro study

Archives of Rheumatology ◽

10.46497/archrheumatol.2022.9109 ◽

2021 ◽

Author(s):

Wen Ting Hao ◽

Lu Huang ◽

Wei Pan ◽

Yi Le Ren

Keyword(s):

Quantitative Polymerase Chain Reaction ◽

In Vitro Study ◽

Synovial Fibroblasts ◽

Akt Pathway ◽

Enzyme Linked Immunosorbent Assay ◽

Mrna Levels ◽

Necrosis Factor Alpha ◽

Messenger Ribonucleic Acid ◽

Tnf Α

Objectives: In this study, we aimed to investigate whether glutathione (GSH) could decrease the secretion of reactive oxygen species (ROS), reduce inflammation, and modulate the phosphatase and tensin homolog deleted on chromosome 10/phosphatidylinositol 3-kinase/AKT (PTEN/PI3K/AKT) in synovial fibroblasts (SFs). Patients and methods: A total of 30 DBA/1J female mice were used in this study. The release of ROS in MH7A cells was examined using a ROS assay kit. The effects of GSH on the messenger ribonucleic acid (mRNA) expression and protein levels of inflammatory cytokines were determined via reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and enzyme-linked immunosorbent assay (ELISA) in mouse SFs and MH7A cells, respectively. The PTEN/PI3K/AKT pathway was investigated via Western blotting. The effects of buthionine-sulfoximine (BSO), as an inhibitor of GSH, on these molecules were examined. Results: The ROS were decreased after GSH treatment, and the mRNA levels of tumor necrosis factor-alpha (TNF-α), interleukin (IL)-1β, IL-6, matrix metalloproteinase (MMP)-1, MMP-3, were also significantly inhibited after GSH stimulation. However, the IL-10 levels were enhanced, and GSH increased the expression of PTEN. The GSH suppressed the activation of phosphorylated (p)-PI3K and p-AKT. The supplementation of the BSO restored the activation of PI3K/AKT pathway with a high production of ROS. The levels of TNF-α, IL-1β and IL-6 were also elevated, when the BSO was added. Conclusion: These findings suggest that GSH can act as an inflammatory suppressor by downregulating the PTEN/PI3K/AKT pathway in MH7A cells. These data indicated a novel function of GSH for improving the inflammation of RA SFs and may help to alleviate the pathological process of RA.

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Inflammatory Cytokine Levels After Endovascular Therapy in Patients With Peripheral Artery Disease

Angiology ◽

10.1177/0003319716682122 ◽

2016 ◽

Vol 68 (8) ◽

pp. 734-740 ◽

Cited By ~ 2

Author(s):

Yasushi Ueki ◽

Takashi Miura ◽

Yusuke Miyashita ◽

Souichiro Ebisawa ◽

Hirohiko Motoki ◽

...

Keyword(s):

Peripheral Artery Disease ◽

Endovascular Therapy ◽

Inflammatory Cytokine ◽

Enzyme Linked Immunosorbent Assay ◽

Peripheral Artery ◽

Stent Restenosis ◽

Tnf Α ◽

Cytokine Levels ◽

Artery Disease ◽

In Stent Restenosis

We evaluated the impact of endovascular therapy (EVT) on inflammatory cytokine levels and its relationship with in-stent restenosis in patients with peripheral artery disease. The study prospectively enrolled 35 patients with intermittent claudication who underwent EVT of the iliofemoral artery. Levels of interleukin 6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), and tumor necrosis factor α (TNF-α) were measured using enzyme-linked immunosorbent assay before and at 2 hours, 4 hours, and 3 months after EVT. All cytokine levels increased significantly after EVT (IL-6 [pg/mL]: from 1.51 [0.84-1.93] before EVT to 6.97 [4.05-20.41] at 2 hours and 13.29 [4.57-31.88] at 4 hours; MCP-1 [pg/mL]: from 326.65 [265.60-406.55] before EVT to 411.18 [341.21-566.27] at 2 hours and 519.36 [383.58-644.85] at 4 hours; TNF-α [pg/mL]: from 1.08 [0.77-1.29] before EVT to 1.25 [0.94-1.81] at 2 hours and 1.27 [0.95-1.59] at 4 hours, all P < .001). However, cytokine levels did not differ significantly between lesions with and without in-stent restenosis. Overall, our results suggest that EVT significantly increases IL-6, MCP-1, and TNF-α levels in the ischemic leg, but this effect is not associated with a higher rate of in-stent restenosis.

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Novel Findings of Anti-cancer Property of Propofol

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520617666170912120327 ◽

2018 ◽

Vol 18 (2) ◽

pp. 156-165 ◽

Cited By ~ 8

Author(s):

Jiaqiang Wang ◽

Chien-shan Cheng ◽

Yan Lu ◽

Xiaowei Ding ◽

Minmin Zhu ◽

...

Keyword(s):

General Anesthesia ◽

Cancer Progression ◽

Molecular Mechanisms ◽

Intravenous Anesthetic ◽

The Past ◽

Anti Cancer ◽

Underlying Mechanisms ◽

Recent Attention

Background: Propofol, a widely used intravenous anesthetic agent, is traditionally applied for sedation and general anesthesia. Explanation: Recent attention has been drawn to explore the effect and mechanisms of propofol against cancer progression in vitro and in vivo. Specifically, the proliferation-inhibiting and apoptosis-inducing properties of propofol in cancer have been studied. However, the underlying mechanisms remain unclear. Conclusion: This review focused on the findings within the past ten years and aimed to provide a general overview of propofol's malignance-modulating properties and the potential molecular mechanisms.

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The impact of FGF19/FGFR4 signaling inhibition in antitumor activity of multi-kinase inhibitors in hepatocellular carcinoma

Scientific Reports ◽

10.1038/s41598-021-84117-9 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Hiroaki Kanzaki ◽

Tetsuhiro Chiba ◽

Junjie Ao ◽

Keisuke Koroki ◽

Kengo Kanayama ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Molecular Mechanisms ◽

Kinase Inhibitors ◽

Progression Free Survival ◽

Erk Signaling ◽

Enzyme Linked Immunosorbent Assay ◽

Antitumor Effects ◽

The Impact

AbstractFGF19/FGFR4 autocrine signaling is one of the main targets for multi-kinase inhibitors (MKIs). However, the molecular mechanisms underlying FGF19/FGFR4 signaling in the antitumor effects to MKIs in hepatocellular carcinoma (HCC) remain unclear. In this study, the impact of FGFR4/ERK signaling inhibition on HCC following MKI treatment was analyzed in vitro and in vivo assays. Serum FGF19 in HCC patients treated using MKIs, such as sorafenib (n = 173) and lenvatinib (n = 40), was measured by enzyme-linked immunosorbent assay. Lenvatinib strongly inhibited the phosphorylation of FRS2 and ERK, the downstream signaling molecules of FGFR4, compared with sorafenib and regorafenib. Additional use of a selective FGFR4 inhibitor with sorafenib further suppressed FGFR4/ERK signaling and synergistically inhibited HCC cell growth in culture and xenograft subcutaneous tumors. Although serum FGF19high (n = 68) patients treated using sorafenib exhibited a significantly shorter progression-free survival and overall survival than FGF19low (n = 105) patients, there were no significant differences between FGF19high (n = 21) and FGF19low (n = 19) patients treated using lenvatinib. In conclusion, robust inhibition of FGF19/FGFR4 is of importance for the exertion of antitumor effects of MKIs. Serum FGF19 levels may function as a predictive marker for drug response and survival in HCC patients treated using sorafenib.

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Surfactant protein A reduces TLR4 and inflammatory cytokine mRNA levels in neonatal mouse ileum

Scientific Reports ◽

10.1038/s41598-021-82219-y ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Lidan Liu ◽

Chaim Z. Aron ◽

Cullen M. Grable ◽

Adrian Robles ◽

Xiangli Liu ◽

...

Keyword(s):

Proinflammatory Cytokine ◽

Inflammatory Cytokine ◽

Protein A ◽

Surfactant Protein ◽

Surfactant Protein A ◽

Mrna Levels ◽

Wild Type ◽

Cytokine Mrna ◽

Protein Levels ◽

Cytokine Levels

AbstractLevels of intestinal toll-like receptor 4 (TLR4) impact inflammation in the neonatal gastrointestinal tract. While surfactant protein A (SP-A) is known to regulate TLR4 in the lung, it also reduces intestinal damage, TLR4 and inflammation in an experimental model of necrotizing enterocolitis (NEC) in neonatal rats. We hypothesized that SP-A-deficient (SP-A−/−) mice have increased ileal TLR4 and inflammatory cytokine levels compared to wild type mice, impacting intestinal physiology. We found that ileal TLR4 and proinflammatory cytokine levels were significantly higher in infant SP-A−/− mice compared to wild type mice. Gavage of neonatal SP-A−/− mice with purified SP-A reduced ileal TLR4 protein levels. SP-A reduced expression of TLR4 and proinflammatory cytokines in normal human intestinal epithelial cells (FHs74int), suggesting a direct effect. However, incubation of gastrointestinal cell lines with proteasome inhibitors did not abrogate the effect of SP-A on TLR4 protein levels, suggesting that proteasomal degradation is not involved. In a mouse model of experimental NEC, SP-A−/− mice were more susceptible to intestinal stress resembling NEC, while gavage with SP-A significantly decreased ileal damage, TLR4 and proinflammatory cytokine mRNA levels. Our data suggests that SP-A has an extrapulmonary role in the intestinal health of neonatal mice by modulating TLR4 and proinflammatory cytokines mRNA expression in intestinal epithelium.

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Molecular Mechanisms of ZC3H12C/Reg-3 Biological Activity and Its Involvement in Psoriasis Pathology

International Journal of Molecular Sciences ◽

10.3390/ijms22147311 ◽

2021 ◽

Vol 22 (14) ◽

pp. 7311

Author(s):

Mateusz Wawro ◽

Jakub Kochan ◽

Weronika Sowinska ◽

Aleksandra Solecka ◽

Karolina Wawro ◽

...

Keyword(s):

Family Member ◽

Cellular Localization ◽

Molecular Mechanisms ◽

Inflammatory Diseases ◽

Association Studies ◽

Psoriasis Patient ◽

Genome Wide Association Studies ◽

Mrna Levels ◽

Transcript Levels

The members of the ZC3H12/MCPIP/Regnase family of RNases have emerged as important regulators of inflammation. In contrast to Regnase-1, -2 and -4, a thorough characterization of Regnase-3 (Reg-3) has not yet been explored. Here we demonstrate that Reg-3 differs from other family members in terms of NYN/PIN domain features, cellular localization pattern and substrate specificity. Together with Reg-1, the most comprehensively characterized family member, Reg-3 shared IL-6, IER-3 and Reg-1 mRNAs, but not IL-1β mRNA, as substrates. In addition, Reg-3 was found to be the only family member which regulates transcript levels of TNF, a cytokine implicated in chronic inflammatory diseases including psoriasis. Previous meta-analysis of genome-wide association studies revealed Reg-3 to be among new psoriasis susceptibility loci. Here we demonstrate that Reg-3 transcript levels are increased in psoriasis patient skin tissue and in an experimental model of psoriasis, supporting the immunomodulatory role of Reg-3 in psoriasis, possibly through degradation of mRNA for TNF and other factors such as Reg-1. On the other hand, Reg-1 was found to destabilize Reg-3 transcripts, suggesting reciprocal regulation between Reg-3 and Reg-1 in the skin. We found that either Reg-1 or Reg-3 were expressed in human keratinocytes in vitro. However, in contrast to robustly upregulated Reg-1 mRNA levels, Reg-3 expression was not affected in the epidermis of psoriasis patients. Taken together, these data suggest that epidermal levels of Reg-3 are negatively regulated by Reg-1 in psoriasis, and that Reg-1 and Reg-3 are both involved in psoriasis pathophysiology through controlling, at least in part different transcripts.

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In vitro responses to platelet-rich-plasma are associated with variable clinical outcomes in patients with knee osteoarthritis

Scientific Reports ◽

10.1038/s41598-021-90174-x ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Habib Zahir ◽

Bijan Dehghani ◽

Xiaoning Yuan ◽

Yurii Chinenov ◽

Christine Kim ◽

...

Keyword(s):

Knee Osteoarthritis ◽

Clinical Outcomes ◽

Inflammatory Responses ◽

Platelet Rich Plasma ◽

Autologous Blood ◽

Limited Information ◽

Knee Oa ◽

Patient Reported ◽

Treatment Comparisons

AbstractAutologous blood-derived products such as platelet-rich plasma (PRP) are widely used to treat musculoskeletal conditions, including knee osteoarthritis (OA). However, the clinical outcomes after PRP administration are often variable, and there is limited information about the specific characteristics of PRP that impact bioactivity and clinical responses. In this study, we aimed to develop an integrative workflow to evaluate responses to PRP in vitro, and to assess if the in vitro responses to PRP are associated with the PRP composition and clinical outcomes in patients with knee OA. To do this, we used a coculture system of macrophages and fibroblasts paired with transcriptomic analyses to comprehensively characterize the modulation of inflammatory responses by PRP in vitro. Relying on patient-reported outcomes and achievement of minimal clinically important differences in OA patients receiving PRP injections, we identified responders and non-responders to the treatment. Comparisons of PRP from these patient groups allowed us to identify differences in the composition and in vitro activity of PRP. We believe that our integrative workflow may enable the development of targeted approaches that rely on PRP and other orthobiologics to treat musculoskeletal pathologies.

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