MAPK regulate MMP-9 gene expression induced by IL-1β and TNF-α in cytotrophoblastic cells

The urgency of the search for inexpensive and effective drugs with localized action for the treatment of rheumatoid arthritis continues unabated. In this study, for the first time we investigated the Cytos-11 antisense oligonucleotide suppression of TNF-α gene expression in a rat model of rheumatoid arthritis induced by complete Freund’s adjuvant. Cytos-11 has been shown to effectively reduce peripheral blood concentrations of TNF-α, reduce joint inflammation, and reduce pannus development. The results achieved following treatment with the antisense oligonucleotide Cytos-11 were similar to those of adalimumab (Humira®); they also compared favorably with those results, which provides evidence of the promise of drugs based on antisense technologies in the treatment of this disease.

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A Novel Method Facilitating the Simple and Low-Cost Preparation of Human Osteochondral Slice Explants for Large-Scale Native Tissue Analysis

International Journal of Molecular Sciences ◽

10.3390/ijms22126394 ◽

2021 ◽

Vol 22 (12) ◽

pp. 6394

Author(s):

Jacob Spinnen ◽

Lennard K. Shopperly ◽

Carsten Rendenbach ◽

Anja A. Kühl ◽

Ufuk Sentürk ◽

...

Keyword(s):

Gene Expression ◽

Cell Viability ◽

Large Scale ◽

Marker Gene ◽

Cell Swelling ◽

Tissue Cell ◽

Joint Research ◽

Sample Number ◽

Tnf Α ◽

Novel Method

For in vitro modeling of human joints, osteochondral explants represent an acceptable compromise between conventional cell culture and animal models. However, the scarcity of native human joint tissue poses a challenge for experiments requiring high numbers of samples and makes the method rather unsuitable for toxicity analyses and dosing studies. To scale their application, we developed a novel method that allows the preparation of up to 100 explant cultures from a single human sample with a simple setup. Explants were cultured for 21 days, stimulated with TNF-α or TGF-β3, and analyzed for cell viability, gene expression and histological changes. Tissue cell viability remained stable at >90% for three weeks. Proteoglycan levels and gene expression of COL2A1, ACAN and COMP were maintained for 14 days before decreasing. TNF-α and TGF-β3 caused dose-dependent changes in cartilage marker gene expression as early as 7 days. Histologically, cultures under TNF-α stimulation showed a 32% reduction in proteoglycans, detachment of collagen fibers and cell swelling after 7 days. In conclusion, thin osteochondral slice cultures behaved analogously to conventional punch explants despite cell stress exerted during fabrication. In pharmacological testing, both the shorter diffusion distance and the lack of need for serum in the culture suggest a positive effect on sensitivity. The ease of fabrication and the scalability of the sample number make this manufacturing method a promising platform for large-scale preclinical testing in joint research.

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Modulatory Effects of Osthole on Lipopolysaccharides-Induced Inflammation in Caco-2 Cell Monolayer and Co-Cultures with THP-1 and THP-1-Derived Macrophages

Nutrients ◽

10.3390/nu13010123 ◽

2020 ◽

Vol 13 (1) ◽

pp. 123

Author(s):

Natalia K. Kordulewska ◽

Justyna Topa ◽

Małgorzata Tańska ◽

Anna Cieślińska ◽

Ewa Fiedorowicz ◽

...

Keyword(s):

Gene Expression ◽

Inflammatory Reaction ◽

Wide Spectrum ◽

Cell Monolayer ◽

In Vivo Studies ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Tnf Α

Lipopolysaccharydes (LPS) are responsible for the intestinal inflammatory reaction, as they may disrupt tight junctions and induce cytokines (CKs) secretion. Osthole has a wide spectrum of pharmacological effects, thus its anti-inflammatory potential in the LPS-treated Caco-2 cell line as well as in Caco-2/THP-1 and Caco-2/macrophages co-cultures was investigated. In brief, Caco-2 cells and co-cultures were incubated with LPS to induce an inflammatory reaction, after which osthole (150–450 ng/mL) was applied to reduce this effect. After 24 h, the level of secreted CKs and changes in gene expression were examined. LPS significantly increased the levels of IL-1β, -6, -8, and TNF-α, while osthole reduced this effect in a concentration-dependent manner, with the most significant decrease when a 450 ng/mL dose was applied (p < 0.0001). A similar trend was observed in changes in gene expression, with the significant osthole efficiency at a concentration of 450 ng/μL for IL1R1 and COX-2 (p < 0.01) and 300 ng/μL for NF-κB (p < 0.001). Osthole increased Caco-2 monolayer permeability, thus if it would ever be considered as a potential drug for minimizing intestinal inflammatory symptoms, its safety should be confirmed in extended in vitro and in vivo studies.

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LPS-induced expression of CD14 in the TRIF pathway is epigenetically regulated by sulforaphane in porcine pulmonary alveolar macrophages

Innate Immunity ◽

10.1177/1753425916669418 ◽

2016 ◽

Vol 22 (8) ◽

pp. 682-695 ◽

Cited By ~ 12

Author(s):

Qin Yang ◽

Maren J Pröll ◽

Dessie Salilew-Wondim ◽

Rui Zhang ◽

Dawit Tesfaye ◽

...

Keyword(s):

Gene Expression ◽

Alveolar Macrophages ◽

Methylation Status ◽

Gene Body ◽

Gene Body Methylation ◽

Tnf Α ◽

Pulmonary Alveolar Macrophages ◽

Cluster Of Differentiation ◽

Cd14 Gene

Pulmonary alveolar macrophages (AMs) are important in defense against bacterial lung inflammation. Cluster of differentiation 14 (CD14) is involved in recognizing bacterial lipopolysaccharide (LPS) through MyD88-dependent and TRIF pathways of innate immunity. Sulforaphane (SFN) shows anti-inflammatory activity and suppresses DNA methylation. To identify CD14 epigenetic changes by SFN in the LPS-induced TRIF pathway, an AMs model was investigated in vitro. CD14 gene expression was induced by 5 µg/ml LPS at the time point of 12 h and suppressed by 5 µM SFN. After 12 h of LPS stimulation, gene expression was significantly up-regulated, including TRIF, TRAF6, NF-κB, TRAF3, IRF7, TNF-α, IL-1β, IL-6, and IFN-β. LPS-induced TRAM, TRIF, RIPK1, TRAF3, TNF-α, IL-1β and IFN-β were suppressed by 5 µM SFN. Similarly, DNMT3a expression was increased by LPS but significantly down-regulated by 5 µM SFN. It showed positive correlation of CD14 gene body methylation with in LPS-stimulated AMs, and this methylation status was inhibited by SFN. This study suggests that SFN suppresses CD14 activation in bacterial inflammation through epigenetic regulation of CD14 gene body methylation associated with DNMT3a. The results provide insights into SFN-mediated epigenetic down-regulation of CD14 in LPS-induced TRIF pathway inflammation and may lead to new methods for controlling LPS-induced inflammation in pigs.

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TNF-α and serum induce SKALP/elafin gene expression in human keratinocytes by a p38 MAP kinase-dependent pathway

Archives of Dermatological Research ◽

10.1007/s004030050475 ◽

2000 ◽

Vol 292 (4) ◽

pp. 180-187 ◽

Cited By ~ 68

Author(s):

R. Pfundt ◽

M. Wingens ◽

M. Bergers ◽

M. Zweers ◽

M. Frenken ◽

...

Keyword(s):

Gene Expression ◽

Map Kinase ◽

Human Keratinocytes ◽

P38 Map Kinase ◽

Tnf Α ◽

Dependent Pathway

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Chronic exposure to TNF-α increases airway mucus gene expression

Journal of Allergy and Clinical Immunology ◽

10.1016/j.jaci.2005.08.059 ◽

2005 ◽

Vol 116 (6) ◽

pp. 1256-1263 ◽

Cited By ~ 54

Author(s):

P BUSSE ◽

T ZHANG ◽

K SRIVASTAVA ◽

B LIN ◽

B SCHOFIELD ◽

...

Keyword(s):

Gene Expression ◽

Chronic Exposure ◽

Tnf Α ◽

Airway Mucus

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Effect of cobalt and chromium ions on MMP-1, TIMP-1, and TNF-α gene expression in human U937 macrophages: A role for tyrosine kinases

Biomaterials ◽

10.1016/j.biomaterials.2005.02.013 ◽

2005 ◽

Vol 26 (28) ◽

pp. 5587-5593 ◽

Cited By ~ 30

Author(s):

Li Luo ◽

Alain Petit ◽

John Antoniou ◽

David J. Zukor ◽

Olga L. Huk ◽

...

Keyword(s):

Gene Expression ◽

Tyrosine Kinases ◽

Chromium Ions ◽

Tnf Α

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Cerium-Containing N-Acetyl-6-Aminohexanoic Acid Formulation Accelerates Wound Reparation in Diabetic Animals

Biomolecules ◽

10.3390/biom11060834 ◽

2021 ◽

Vol 11 (6) ◽

pp. 834

Author(s):

Ekaterina Blinova ◽

Dmitry Pakhomov ◽

Denis Shimanovsky ◽

Marina Kilmyashkina ◽

Yan Mazov ◽

...

Keyword(s):

Gene Expression ◽

Wound Healing ◽

Topical Therapy ◽

Skin Defect ◽

Ki 67 ◽

Fgfr3 Gene ◽

White Rats ◽

Tnf Α ◽

Acid Formulation ◽

Healing Property

Background: The main goal of our study was to explore the wound-healing property of a novel cerium-containing N-acethyl-6-aminohexanoate acid compound and determine key molecular targets of the compound mode of action in diabetic animals. Methods: Cerium N-acetyl-6-aminohexanoate (laboratory name LHT-8-17) as a 10 mg/mL aquatic spray was used as wound experimental topical therapy. LHT-8-17 toxicity was assessed in human skin epidermal cell culture using (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. A linear wound was reproduced in 18 outbred white rats with streptozotocin-induced (60 mg/kg i.p.) diabetes; planar cutaneous defect was modelled in 60 C57Bl6 mice with streptozotocin-induced (200 mg/kg i.p.) diabetes and 90 diabetic db/db mice. Firmness of the forming scar was assessed mechanically. Skin defect covering was histologically evaluated on days 5, 10, 15, and 20. Tissue TNF-α, IL-1β and IL-10 levels were determined by quantitative ELISA. Oxidative stress activity was detected by Fe-induced chemiluminescence. Ki-67 expression and CD34 cell positivity were assessed using immunohistochemistry. FGFR3 gene expression was detected by real-time PCR. LHT-8-17 anti-microbial potency was assessed in wound tissues contaminated by MRSA. Results: LHT-8-17 4 mg twice daily accelerated linear and planar wound healing in animals with type 1 and type 2 diabetes. The formulated topical application depressed tissue TNF-α, IL-1β, and oxidative reaction activity along with sustaining both the IL-10 concentration and antioxidant capacity. LHT-8-17 induced Ki-67 positivity of fibroblasts and pro-keratinocytes, upregulated FGFR3 gene expression, and increased tissue vascularization. The formulation possessed anti-microbial properties. Conclusions: The obtained results allow us to consider the formulation as a promising pharmacological agent for diabetic wound topical treatment.

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