scholarly journals A Phytocomplex Consisting of Tropaeolum majus L. and Salvia officinalis L. Extracts Alleviates the Inflammatory Response of Dermal Fibroblasts to Bacterial Lipopolysaccharides

2020 ◽  
Vol 2020 ◽  
pp. 1-14 ◽  
Author(s):  
Tunde Jurca ◽  
Ioana Baldea ◽  
Gabriela Adriana Filip ◽  
Diana Olteanu ◽  
Simona Clichici ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Inflammatory Response ◽  
Dermal Fibroblast ◽  
Physicochemical Characterization ◽  
Antibacterial Activities ◽  
Dermal Fibroblasts ◽  
High Dose ◽  
Cytokine Levels ◽  
Bacterial Lipopolysaccharides

Background. The antimicrobial activity and effects of a phytocomplex consisting of Tropaeolum flos (T) and Salviae folium (S) extracts on the cytokine levels and transcription factors on dermal fibroblast BJ exposed to bacterial lipopolysaccharides were examined. Methods. In order to select the most optimal combination ratio of the two extracts for using in vitro, the physicochemical characterization of vegetal extract mixtures was performed and the antioxidant and antibacterial activities were evaluated on five different formulations of T : S, namely, 1 : 1, 1 : 2, 2 : 1, 3 : 1, and 1 : 3. The best combination of bioactive compounds with regard to antioxidant and antibacterial activities (T : S 1 : 2) was selected for in vitro evaluation of the anti-inflammatory effect. Human dermal fibroblast BJ cells were treated with two doses of the extract mixture and then exposed to bacterial lipopolysaccharides (LPS). The levels of the cytokines involved in inflammatory response, namely, interleukin- (IL-) 6, tumor necrosis factor- (TNF-) α, IL-31, and IL-33, were quantified by ELISA, and the expression of transcription factors, namely, signal transducer and activator of transcription (STAT) 3, nuclear factor kappa B (NFκB), and phosphorylated NFκB (pNFκB), were evaluated by western blot analysis. Results. The results have shown that the mixture of T : S 1 : 2 exhibited significant antibacterial effects on Staphylococcus aureus ATCC 25923. LPS exposure increased the cytokine levels in BJ cells and enhanced the NFκB expression. The pretreatment of BF cells exposed to LPS with the two doses of the extract mixture markedly inhibited the increase of IL-33 and TNF-α levels and amplified the NFκB expression and its activation, especially with the high dose. The low doses of the extract reduced NFκB expression but increased its activation. Conclusions. These experimental findings suggest that the mixture of T : S 1 : 2 can exert some protection against bacterial infections and inflammation induced by LPS in BJ cells being a good therapeutic option in related conditions associated with inflammation.

scholarly journals In VitroActivities of Daptomycin-, Vancomycin-, and Teicoplanin-Loaded Polymethylmethacrylate against Methicillin-Susceptible, Methicillin-Resistant, and Vancomycin-Intermediate Strains of Staphylococcus aureus

2011 ◽  
Vol 55 (12) ◽  
pp. 5480-5484 ◽  
Author(s):  
Yuhan Chang ◽  
Wen-Chien Chen ◽  
Pang-Hsin Hsieh ◽  
Dave W. Chen ◽  
Mel S. Lee ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
High Performance ◽  
Low Dose ◽  
Antibacterial Activities ◽  
High Dose ◽  
Bone Cements ◽  
Antibacterial Effects ◽  
Methicillin Resistant ◽  
Content Type

ABSTRACTThe objective of this study was to evaluate the antibacterial effects of polymethylmethacrylate (PMMA) bone cements loaded with daptomycin, vancomycin, and teicoplanin against methicillin-susceptibleStaphylococcus aureus(MSSA), methicillin-resistantStaphylococcus aureus(MRSA), and vancomycin-intermediateStaphylococcus aureus(VISA) strains. Standardized cement specimens made from 40 g PMMA loaded with 1 g (low-dose), 4 g (middle-dose) or 8 g (high-dose) antibiotics were tested for elution characteristics and antibacterial activities. The patterns of release of antibiotics from the cement specimens were evaluated usingin vitrobroth elution assay with high-performance liquid chromatography. The activities of broth elution fluid against differentStaphylococcus aureusstrains (MSSA, MRSA, and VISA) were then determined. The antibacterial activities of all the tested antibiotics were maintained after being mixed with PMMA. The cements loaded with higher dosages of antibiotics showed longer elution periods. Regardless of the antibiotic loading dose, the teicoplanin-loaded cements showed better elution efficacy and provided longer inhibitory periods against MSSA, MRSA, and VISA than cements loaded with the same dose of vancomycin or daptomycin. Regarding the choice of antibiotics for cement loading in the treatment ofStaphylococcus aureusinfection, teicoplanin was superior in terms of antibacterial effects.

Haloperidol and Risperidone at high concentrations activate an in vitro inflammatory response of RAW 264.7 macrophage cells by induction of apoptosis and modification of cytokine levels

2015 ◽  
Vol 233 (9) ◽  
pp. 1715-1723 ◽  
Author(s):  
Ivo Emílio da Cruz Jung ◽  
Alencar Kolinski Machado ◽  
Ivana Beatrice Mânica da Cruz ◽  
Fernanda Barbisan ◽  
Verônica Farina Azzolin ◽  
...  
Keyword(s):  
Inflammatory Response ◽  
Raw 264.7 ◽  
Macrophage Cells ◽  
Raw 264.7 Macrophage ◽  
Cytokine Levels ◽  
Raw 264.7 Macrophage Cells ◽  
Induction Of Apoptosis ◽  
High Concentrations

scholarly journals Identification of a Functional Peroxisome Proliferator-Activated Receptor Response Element in the Rat Catalase Promoter

2002 ◽  
Vol 16 (12) ◽  
pp. 2793-2801 ◽  
Author(s):  
Geoffrey D. Girnun ◽  
Frederick E. Domann ◽  
Steven A. Moore ◽  
Mike E. C. Robbins
Keyword(s):  
Reactive Oxygen Species ◽  
Transcription Factors ◽  
Inflammatory Response ◽  
Reactive Oxygen ◽  
Peroxisome Proliferator ◽  
Oxygen Species ◽  
Response Element ◽  
Peroxisome Proliferator Activated Receptor ◽  
Promoter Deletion Analysis

Abstract Peroxisomal proliferator-activated receptor (PPAR)γ has been shown to decrease the inflammatory response via transrepression of proinflammatory transcription factors. However, the identity of PPARγ responsive genes that decrease the inflammatory response has remained elusive. Because generation of the reactive oxygen species hydrogen peroxide (H2O2) plays a role in the inflammatory process and activation of proinflammatory transcription factors, we wanted to determine whether the antioxidant enzyme catalase might be a PPARγ target gene. We identified a putative PPAR response element (PPRE) containing the canonical direct repeat 1 motif, AGGTGA-A-AGTTGA, in the rat catalase promoter. In vitro translated PPARγ and retinoic X receptor-α proteins were able to bind to the catalase PPRE. Promoter deletion analysis revealed that the PPRE was functional, and a heterologous promoter construct containing a multimerized catalase PPRE demonstrated that the PPRE was necessary and sufficient for PPARγ-mediated activation. Treatment of microvascular endothelial cells with PPARγ ligands led to increases in catalase mRNA and activity. These results demonstrate that PPARγ can alter catalase expression; this occurs via a PPRE in the rat catalase promoter. Thus, in addition to transrepression of proinflammatory transcription factors, PPARγ may also be modulating catalase expression, and hence down-regulating the inflammatory response via scavenging of reactive oxygen species.

Antibacterial Wound Healing Gels from Oligochitosan Salts

2012 ◽  
Vol 506 ◽  
pp. 31-34
Author(s):  
W. Janvikul ◽  
P. Ngamviriyavong ◽  
P. Uppanun ◽  
P. Tanjak ◽  
N. Sangjun
Keyword(s):  
Staphylococcus Epidermidis ◽  
Wound Closure ◽  
Collagen Gel ◽  
Antibacterial Activities ◽  
Dermal Fibroblasts ◽  
Collagen Gels ◽  
Collagen Gel Matrix ◽  
Growth Of Bacteria

Oligochitosan salt-based antibacterial wound gels were developed and evaluated in both in vitro and in vivo models. The antibacterial activities of the oligochitosan salts and the wound gels were investigated against Staphylococcus epidermidis RP625 and Escherichia coli ATCC 11775. The minimum inhibitory concentrations (MIC) of the oligochitosan salts were found in the range of 16-256 μg/mL. The wound gels demonstrated their in vitro activities on inhibiting the growth of bacteria. The 3-D collagen gel matrix containing human dermal fibroblasts cultured with each test gel was used as an in vitro model for the examination of cell proliferation and secretion of interleukin-8 (IL-8). The gels appeared to promote the proliferation and formation of cellular process of the fibroblasts in the 3-D collagen gels and stimulate the fibroblasts to produce more IL-8. In the in vivo model, it was noted that the gels could accelerate the wound closure process. The wounds were completely closed within 14 days.

scholarly journals Contribution of IL-38 in Lung Immunity During Pseudomonas Aeruginosa-Induced Pneumonia

2021 ◽  
Author(s):  
Qiang Wei ◽  
Xi Chen ◽  
Chuanjiang Wang
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Inflammatory Response ◽  
Murine Model ◽  
Cell Apoptosis ◽  
Protective Role ◽  
Bacterial Removal ◽  
Cytokine Levels ◽  
New Type ◽  
Treg Differentiation

Abstract Objective: Interleukin-38 (IL-38), a new type of cytokine, is involved in processes such as tissue repair, inflammatory response, and immune response. However, its function in pneumonia caused by Pseudomonas aeruginosa is still unclear.Methods: In this study, we detected circulating IL-38 in adults affected by pneumonia caused by P. aeruginosa. The P. aeruginosa-induced pneumonia WT murine model was adopted to evaluate the effect of IL-38 on Treg differentiation, cell apoptosis, survival, tissue damage, inflammation, and bacterial removal.Results: IL-38 is insufficiently secreted in patients who died of P.A. pneumonia.Recombinant IL-38 improved survival, whereas anti-IL-38 antibody reduced survival in the experimental pneumonia murine model. IL-38 exposure reduced the inflammatory response, as suggested by the lung injury, and reduced cytokine levels (IL-1β, IL-6, IL-17A, TNF-α, and CXCL-1, but not IL-10). It also increased bacterial clearance and reduced cell apoptosis in the lungs. Furthermore, IL-38 was shown to reduce TBK1 expression in vitro when naïve CD4+ T lymphocytes were differentiated to Tregs and played a protective role in P.A. pneumonia.Conclusions: To summarize, the above findings provide additional insights into the mechanism of IL-38 in the treatment of P.A. pneumonia.

scholarly journals In Vitro and In Vivo Activity of AS101 against Carbapenem-Resistant Acinetobacter baumannii

2021 ◽  
Vol 14 (8) ◽  
pp. 823
Author(s):  
Tsung-Ying Yang ◽  
Sung-Pin Tseng ◽  
Heather Nokulunga Dlamini ◽  
Po-Liang Lu ◽  
Lin Lin ◽  
...  
Keyword(s):  
Acinetobacter Baumannii ◽  
Antibacterial Activities ◽  
Treated Group ◽  
Infection Model ◽  
High Dose ◽  
Mouse Infection Model ◽  
Carbapenem Resistant ◽  
Time Kill

The increasing trend of carbapenem-resistant Acinetobacter baumannii (CRAB) worldwide has become a concern, limiting therapeutic alternatives and increasing morbidity and mortality rates. The immunomodulation agent ammonium trichloro (dioxoethylene-O,O′-) tellurate (AS101) was repurposed as an antimicrobial agent against CRAB. Between 2016 and 2018, 27 CRAB clinical isolates were collected in Taiwan. The in vitro antibacterial activities of AS101 were evaluated using broth microdilution, time-kill assay, reactive oxygen species (ROS) detection and electron microscopy. In vivo effectiveness was assessed using a sepsis mouse infection model. The MIC range of AS101 for 27 CRAB isolates was from 0.5 to 32 µg/mL, which is below its 50% cytotoxicity (approximately 150 µg/mL). Bactericidal activity was confirmed using a time-kill assay. The antibacterial mechanism of AS101 was the accumulation of the ROS and the disruption of the cell membrane, which, in turn, results in cell death. The carbapenemase-producing A. baumannii mouse sepsis model showed that AS101 was a better therapeutic effect than colistin. The mice survival rate after 120 h was 33% (4/12) in the colistin-treated group and 58% (7/12) in the high-dose AS101 (3.33 mg/kg/day) group. Furthermore, high-dose AS101 significantly decreased bacterial population in the liver, kidney and spleen (all p < 0.001). These findings support the concept that AS101 is an ideal candidate for further testing in future studies.

The Effect of Cyclosporine A on Dermal Fibroblast Cell - Transcriptomic Analysis of Inflammatory Response Pathway

2020 ◽  
Vol 21 (12) ◽  
pp. 1213-1223
Author(s):  
Grażyna Janikowska ◽  
Ewa Kurzeja ◽  
Marcin Janikowski ◽  
Barbara Strzałka-Mrozik ◽  
Alina Pyka-Pająk ◽  
...  
Keyword(s):  
Inflammatory Response ◽  
Cyclosporine A ◽  
Dermal Fibroblast ◽  
Dermal Fibroblasts ◽  
Transcriptional Level ◽  
Main Task ◽  
Human Dermal Fibroblasts ◽  
Therapeutic Concentration ◽  
Cutaneous Manifestations ◽  
Sites Of Action

Background: The first immunosuppressive drug - cyclosporine A (CsA) has many unquestioned merits in maintaining organ transplants in patients, as well as, in the treatment of many inflammatory diseases, also associated with cutaneous manifestations. The main task of this drug is to suppress the inflammatory response at the sites of action, which is not well known. Objective: The objective of this study was to evaluate the influence of CsA in therapeutic concentration on the expression of genes associated with the inflammatory response pathway in normal human dermal fibroblasts (NHDF; CC-2511), and this study attempted to determine the mechanism of its action. Methods: The cytotoxicity MTT test was performed. The expression of the inflammatory response pathway genes was determined using HG-U133A_2.0 oligonucleotide microarrays. Statistical analysis was performed by GeneSpring 13.0 software using the PL-Grid platform. Results: Among the 5,300 mRNA, only 573 were changed significantly in response to CsA compared to the control fibroblasts (P≤0.05). CsA inhibited the expression of most genes associated with the inflammatory response in NHDFs. There were only 19 genes with a fold change (FC) lower than -2.0, among which EGR1, FOS, PBK, CDK1 and TOP2A had the lowest expression, as did CXCL2 which can directly impact inflammation. Furthermore, ZNF451 was strongly induced, and COL1A1, COL3A1, IL33, TNFRSFs were weakly up-regulated (FC lower than 2.0). Conclusion: The CsA in therapeutic concentration influences the genes linked to the inflammatory response (in the transcriptional level) in human dermal fibroblasts. The findings suggest that the potential mechanism of CsA action in this concentration and on these genes can be associated with a profibrotic and proapoptotic, and genotoxic effects.

Evaluation of Gene Expression and In Vitro Enzyme Study for Antiaging Effect of Crocin and Lutein

2020 ◽  
Vol 10 (5) ◽  
pp. 595-604
Author(s):  
Archana A. Naik ◽  
Chhaya H. Gadgoli ◽  
Arvind B. Naik
Keyword(s):  
Gene Expression ◽  
Cell Line ◽  
Dermal Fibroblast ◽  
Dermal Fibroblasts ◽  
Major Constituent ◽  
Chromatographic Technique ◽  
Gene Expression Study ◽  
Expression Study ◽  
Enzyme Study

Background: Tubular calyx of flowers of Nyctanthes arbour-tristis contains an apocarotenoid crocin, a major constituent present in saffron stigma. The flowers of N. arbortristis are readily available, hence can be an economic substitute for saffron. Lutein from flowers of Tagetes patula, is another carotenoid which is a popular antioxidant. Objective: Oxidative stress is a major contributor to the process of aging. Carotenoids are powerful antioxidants. Hence, the study was carried out to evaluate anticollagenase activity and antielastase activity using gene expression study in Human dermal fibroblasts. Methods: Crocin was isolated from the tubular calyx of Nyctanthes arbortristis using flash chromatographic technique and lutein was isolated using column chromatography. Anticollagenase and antielastase activity of crocin and lutein were carried out using collagenase from Clostridium histolyticum as enzyme and porcine pancreatic elastase. Cytotoxicity of crocin and lutein was determined in Human Dermal Fibroblast cell line (HDF) through MTT assay. In gene expression study, the HDF Cell line was inoculated with Crocin (450 and 250 ppm) and lutein (100 and 50 ppm) separately for 24 hrs and the m-RNA expression levels of COL Type-1 and elastin were determined using RT-PCR. The results were compared with standards. Result: Crocin and lutein both showed inhibition of collagenase and elastase enzyme which are responsible for aging process. The cytotoxic concentration CTC 50 (ppm) for Crocin and lutein was found to be 790.2 ppm and 137.14 ppm. Gene expression study on crocin rich extract of Nyctanthes arbortristis showed upregulation of both collagen and elastin gene whereas lutein rich extract having concentration100 μg/ml showed up regulation by 0.02 fold and concentration 50 μg/ml showed down regulation. Conclusion: In vitro collagenase and elastase enzyme study and Gene expression study showed that these carotenoids are potential antiageing agents which can be substituted to synthetic cosmeceuticals as well as saffron.

scholarly journals Enhancement Of Dermal Fibroblast Isolation Method

2015 ◽  
Vol 16 (1) ◽  
pp. 65-69
Author(s):  
Milan Zaric ◽  
Ivana Nikolic ◽  
Ivanka Zelen ◽  
Marina Mitrovic ◽  
Zoran Milosavljevic
Keyword(s):  
Dermal Fibroblast ◽  
Petri Dish ◽  
Culture Conditions ◽  
Dermal Fibroblasts ◽  
Enzyme Digestion ◽  
Digestion Method ◽  
Primary Isolation ◽  
Donor Cells ◽  
Cell Culture Conditions

ABSTRACTCultivated fibroblasts have been widely used in a large number of in vitro studies. Although they readily proliferate under cell culture conditions, improvements in methods for their isolation are necessary. Here, we present our modified enzyme digestion method and compare its efficiency with commonly used techniques.Three foreskin samples from young, middle-aged and old donors were used. The classical explant, standard enzyme digestion method with collagenase and our improved enzyme digestion method were compared for efficiency of fibroblast isolation and the time needed to achieve 95% confluence in a 30-mm Petri dish.The explant method was the slowest to achieve fibroblast confluence, especially with the tissues from the older donors (up to 23 days). With the standard enzyme digestion method, the skin tissue was partially digested, but the fibroblasts reached confluence much faster (the younger donor cells needed approximately 7 days to reach confluence). Our modified “mixed” enzyme digestion method was the fastest (the fibroblasts from the younger donors needed up to 5 days to reach confluence).For studies requiring the primary isolation and cultivation of dermal fibroblasts, the best method to achieve this goal is the tissue digestion method with the multiple enzyme solution.

scholarly journals Information-dependent Enrichment Analysis Reveals Time-dependent Transcriptional Regulation of the Estrogen Pathway of Toxicity

2016 ◽  
Author(s):  
Salil N. Pendse ◽  
Alexandra Maertens ◽  
Michael Rosenberg ◽  
Dipanwita Roy ◽  
Rick A. Fasani ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Transcription Factors ◽  
Expression Patterns ◽  
Enrichment Analysis ◽  
Fold Change ◽  
Gene Set Enrichment Analysis ◽  
High Dose ◽  
Functional Responses ◽  
Pathways Of Toxicity

The twenty-first century vision for toxicology involves a transition away from high-dose animal studies and into in vitro and computational models. This movement requires mapping pathways of toxicity through an understanding of how in vitro systems respond to chemical perturbation. Uncovering transcription factors responsible for gene expression patterns is essential for defining pathways of toxicity, and ultimately, for determining chemical mode of action, through which a toxicant acts. Traditionally this is achieved via chromatin immunoprecipitation studies and summarized by calculating, which transcription factors are statistically associated with the up- and down-regulated genes. These lists are commonly determined via statistical or fold-change cutoffs, a procedure that is sensitive to statistical power and may not be relevant to determining transcription factor associations. To move away from an arbitrary statistical or fold-change based cutoffs, we have developed in the context of the Mapping the Human Toxome project, a novel enrichment paradigm called Information Dependent Enrichment Analysis (IDEA) to guide identification of the transcription factor network. We used the test case of endocrine disruption of MCF-7 cells activated by 17β estradiol (E2). Using this new approach, we were able to establish a time course for transcriptional and functional responses to E2. ERα and ERβ are associated with short-term transcriptional changes in response to E2. Sustained exposure leads to the recruitment of an additional ensemble of transcription factors and alteration of cell-cycle machinery. TFAP2C and SOX2 were the transcription factors most highly correlated with dose. E2F7, E2F1 and Foxm1, which are involved in cell proliferation, were enriched only at 24h. IDEA is, therefore, a novel tool to identify candidate pathways of toxicity, clearly outperforming Gene-set Enrichment Analysis but with similar results as Weighted Gene Correlation Network Analysis, which helps to identify genes not annotated to pathways.

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