Use In Inflammatory Disorders Research Articles

Longevity-associated BPIFB4 gene counteracts the inflammatory signaling

BackgroundIncreased levels of pro-inflammatory proteins in plasma can be detected in older individuals and associate with the so called chronic low-grade inflammation, which contributes to a faster progression of aged-related cardiovascular (CV) diseases, including frailty, neurodegeneration, gastro-intestinal diseases and disorders reflected by alterations in the composition of gut microbiota. However, successful genetic programme of long-living individuals alters the trajectory of the ageing process, by promoting an efficient immune response that can counterbalance deleterious effects of inflammation and the CV complications. This is the case of BPIFB4 gene in which, homozygosity for a four single-nucleotide polymorphism (SNP) haplotype, the Longevity-Associated Variant (LAV) correlates with prolonged health span and reduced risk of CV complications and inflammation. The relation between LAV-BPIFB4 and inflammation has been proven in different experimental models, here we hypothesized that also human homozygous carriers of LAV-BPIFB4 gene may experience a lower inflammatory burden as detected by plasma proteomics that could explain their favourable CV risk trajectory over time. Moreover, we explored the therapeutic effects of LAV-BPIFB4 in inflammatory disease and monolayer model of intestinal barrier.ResultsWe used high-throughput proteomic approach to explore the profiles of circulating proteins from 591 baseline participants selected from the PLIC cohort according to the BPIFB4 genotype to identify the signatures and differences of BPIFB4 genotypes useful for health and disease management. The observational analysis identified a panel of differentially expressed circulating proteins between the homozygous LAV-BPIFB4 carriers and the other alternative BPIFB4 genotypes highlighting in the latter ones a higher grade of immune-inflammatory markers. Moreover, in vitro studies performed on intestinal epithelial organs from inflammatory bowel disease (IBD) patients and monolayer model of intestinal barrier demonstrated the benefit of LAV-BPIFB4 treatment.ConclusionsHomozygosity for LAV-BPIFB4 results in the attenuation of inflammation in PLIC cohort and IBD patients providing preliminary evidences for its therapeutic use in inflammatory disorders that need to be further characterized and confirmed by independent studies.

Evaluation of Phytochemicals and anti-inflammatory effects of hexane extracts from <i>Borreria stachydea</i> [(DC) Hutch. and Dalziel] (Rubiaceae)

This studies evaluated the preliminary phytoconstituents of hexane extract of Borreeia stachydea (Rubiaceae), toxicity studies using OECD guideline, acute and chronic inflammation. This revealed the presence of alkaloids, carbohydrate, steroid, triterpenes, and tannins. Acute toxicity study showed no symptoms of toxicity even at 5000 mg/kg for the period of fourteen days. Its efficacy was significant at doses of 125 at p< 0.01 and 500 mg/kg at p< 0.05 when compared to distilled water treated groups. It is also significant at 125 mg/kg at p< 0.05 in 4 hr. Also 250 and 500 mg/kg at p< 0.001 in 5th hr compared to 1 hr treated group. The standard drug Diclopenac (10 mg /kg) provided a significance difference at (p< 0.001). Also at (4th hour), the extract at 500, 250 and 125 mg/kg inhibited inflammation induced by carrageenan by 92.16, 88.24 and 88. 24 % respectively compared to standard drug that has the inhibition by 90.20 %. Chronic inflammation that served as a long term inflammation of this studies indicated a significance difference at p< 0.05 with 500 mg/kg on day 4, 5 and 6 at p<0.01 with 250 mg/kg on day 5 and 6. Consequently, a significance was also observed at p<0.01 and p<0.001 on day 5 and 6 All compared to day 1 treated group. Acute toxicity study were observed that the extracts did not show any behavioral changes or mortality even at a dose of 5000 mg/kg this indicated its safety and efficacy. However, further studies can be carried out to reveal the exact mechanisms of action responsible to manage inflammation. This study has highlighted that the anti-inflammatory activity of this plant could be a potential new source as well as scientific proof of its ethno-pharmacological use in inflammatory disorders.

Coriandrum sativum improve neuronal function via inhibition of oxidative/nitrosative stress and TNF-α in diabetic neuropathic rats

Ethnophramacological relevanceCoriandrum sativum L. is traditionally acknowledged for its use in inflammatory disorders, altered blood lipid levels, respiratory and digestive problems. Aim of the studyThe present study investigates possible role of hydro-alcoholic extract of C. sativum (CHA) seeds in the attenuation of indices of diabetic peripheral neuropathy (DPN). Materials and methodsPhytochemical analysis was carried out by employing chromatographic, spectroscopic as well as spectrometric techniques. Diabetes was induced by a single i.p. injection of freshly prepared STZ (65 mg/kg). The indexed markers of DPN, i.e., thermal and mechanical hyperalgesia were found to be prominent on the 60th day of STZ administration. Administration of CHA (100, 200, and 400 mg/kg, p.o.) for 30 days was started on the substantiation of DPN onset. Molecular docking study was performed by targeting TNF-α. ResultsPhytochemical analysis revealed the presence of flavonoids, terpenoids, and phenolic acids. Oral administration of CHA considerably attenuated hyperglycemia and decreased pain threshold in diabetic rats as well as modulated oxidative-nitrosative stress. Docking study suggested good affinity of flavonoids when docked into the binding site of TNF-α. ConclusionIn conclusion, using STZ model, it was successfully predicted that CHA might be beneficial in diabetes-induced neuropathic pain by inhibiting oxidative/nitrosative stress and inflammatory cytokine.

Inhibitory Effects of Gynura procumbens Ethanolic Extract on Nitric Oxide Production and Inducible Nitric Oxide Synthase (iNOS) Protein Expression in Macrophages

Nitric oxide (NO) overproduction by inducible nitric oxide synthase (iNOS) may be associated with acute and chronic inflammations. Macrophages as important cells in the innate immune system are able to be stimulated and can lead to iNOS activation and excessive NO production. Gynura procumbens is a medicinal plant traditionally used in treating various ailments including inflammation but the mechanism of anti-inflammatory activity of this plant is still elusive. This study was carried out to investigate the anti-inflammatory therapeutic effects of Gynura procumbens ethanolic extract on NO production and iNOS protein expression in RAW 264.7 macrophages stimulated with lipopolysaccharide (LPS). Cell viability of RAW 264.7 macrophages treated with Gynura procumbens ethanolic extract was determined by MTT assay. NO production was determined by Griess assay following Gynura procumbens ethanolic extract treatment alone or in combination with LPS stimulation. Protein expression of iNOS was determined by western blot. RAW 264.7 macrophages viability of more than 90% was observed after 24 h treatment with Gynura procumbens ethanolic extract concentration range of 3.9 μg/mL to 500 μg/mL. Significant inhibition of NO production level has been identified in LPS-stimulated RAW 264.7 cells pre-treated with 250 μg/mL Gynura procumbens ethanolic extract (p<0.05) while all selected concentrations of Gynura procumbens ethanolic extract showed no significant alteration of NO production in the absence of LPS stimulation. Pre-treatment of 250 μg/mL Gynura procumbens ethanolic extract also demonstrated significant suppression of iNOS protein expression in LPS-stimulated RAW 264.7 cells (p<0.05). In conclusion, this study demonstrates that Gynura procumbens ethanolic extract exhibits anti-inflammatory potential through inhibition of NO production and iNOS protein expression in LPS-stimulated macrophages, suggesting that this plant could be further researched for its beneficial use in inflammatory disorders.

In vitro, fluorescence-quenching and computational studies on the interaction between lipoxygenase and 5-hydroxy-3′,4′,7-trimethoxyflavone from Lippia nodiflora L.

Lippia nodiflora L. is extensively used in traditional medicine for several medicinal purposes, including their use in inflammatory disorders. In this study, the folk use of L. nodiflora was validated using the isolated natural compound, 5-hydroxy-3′,4′,7-trimethoxyflavone (HTMF) by in vitro, fluorescence spectroscopic and molecular modeling studies with lipoxygenase (LOX), because LOX plays an essential role in inflammatory responses. In this perspective, the methanol extract and HTMF are shown to demonstrate prominent inhibitory activity against soybean lipoxygenase, with an IC50 value of 21.12 and 23.97 µg/ml, respectively. The data obtained from the spectroscopic method revealed that the quenching of intrinsic fluorescence of LOX is produced as a result of the complex formation of LOX–HTMF. The binding mode analysis of HTMF within the LOX enzyme suggested that hydrogen bond formation, hydrophobic interaction and π–π stacking could account for the binding of HTMF. Molecular dynamics results indicated the interaction of HTMF with LOX and the stability of ligand–enzyme complex was maintained throughout the simulation. The computational results are reliable with experimental facts and provided a good representation for understanding the binding mode of HTMF inside the active site of lipoxygenase enzyme.

Synthesis, spectral characterization, cytotoxicity and enzyme-inhibiting activity of new ferrocene–indole hybrids

Incorporation of a ferrocene moiety into the 2-phenylindole scaffold has been recently reported to drastically improve the cytotoxic activity of the parent compounds. In our search for new promising cytotoxic agents we designed and prepared two new ferrocene–indole hybrids, 2-(3-ferrocenylphenyl)-1H-indole and 2-(4-ferrocenylphenyl)-1H-indole, utilizing the Fischer indole synthesis as the key step. Detailed spectral analyses, including 1D and 2D NMR in various solvents, have been carried out to corroborate the structures of the synthesized compounds. In this work, we also present the first results of biological studies of these two compounds. Both compounds showed weak anticholinesterase activity but high cytotoxicity against rat peritoneal macrophages and the crustacean Artemia salina. Also, both compounds showed significant myeloperoxidase inhibiting activity, thus suggesting a potential use in inflammatory disorders. The results of these tests are very encouraging as they also suggest possible cytotoxic activities of these compounds against human cancer cell lines.

Bioassay-guided isolation, identification and molecular ligand-target insight of lipoxygenase inhibitors from leaves of Anisomeles malabarica R.Br.

Background:Anisomeles malabarica R. Br. (Lamiaceae) is extensively used in traditional medicine in major parts of India for several medicinal purposes, including their use in rheumatism.Materials and Methods:The air-dried leaves of A. malabarica were extracted with ethanol, defatted with n-hexane and then successively partitioned into chloroform and n-butanol fractions. Bioassay-guided fractionation and purification of chloroform fraction from A. malabarica lead to the isolation of lipoxygenase (LOX) inhibitors. The structures of isolated compounds were elucidated by ultraviolet, infrared, 1H nuclear magnetic resonance (NMR), 13C NMR and mass spectrometry spectroscopic techniques and assessed further by in vitro soybean lipoxygenase (sLOX) assay. In addition, the enzyme type inhibition was evaluated through molecular docking technique as a part of computational study.Results:The bioactive compounds 3, 4 dihydroxy benzoic acid (1) and 4’, 5, 7-trihydroxyflavone (2) were isolated from chloroform fraction of A. malabarica, whose bioactivity was observed to be dose-dependent compared to n-butanol fraction. Among the compounds, 3, 4 dihydroxy benzoic acid showed significant sLOX inhibitory activity with 74.04% ±2.6% followed by 4’, 5, 7-trihydroxyflavone (34.68% ±1.9%). The computational analysis of compounds showed their molecular interaction with important amino acid residues and nonheme iron atom in the catalytic site of LOX by enlightening their potential binding mode at molecular level.Conclusions:The LOX inhibitory constituents were identified from A. malabarica by means of bioassay-guided fractionation process. The results derived from in vitro and computational experiments confirm the potential of the isolated compounds and provide additional evidence for its traditional use in inflammatory disorders.

Molecular docking of Diospyrin as a LOX inhibitory compound

Diospyros lotus is traditionally used in various diseases including inflammation. In the current study an effort has been made to identify a bioactive constituent from D. lotus in order to scientifically validate its use in inflammatory disorders. Diospyrin was isolated from D. lotus and exhibited significant lipoxygenase (LOX) inhibitory activity (IC50 value: 31.89±0.14μmol). Molecular docking revealed significant molecular interactions between Diospyrin and LOX showing promising potential for further optimization as a potential anti-inflammatory lead compound.

Studies on anti-inflammatory and analgesic activities of betel nut in rodents

Areca catechu, commonly known as betel nut, is very famous for its medicinal use in multiple disorders. It is also popular as a remedy against inflammatory disorders in the Unani (Greco-Arab) system of medicine. This study was aimed at investigating the anti-inflammatory and analgesic activities of the crude extract of Areca catechu and its respective fractions. Paw edema, formalin-induced nociception and acetic acid-induced writhing assays were carried out in vivo. Free radical scavenging activity of the plant extract was performed in vitro. Preliminary experiments using a single dose (100 mg/kg) of Areca catechu and its respective fractions demonstrated an anti-inflammatory effect on carrageenan-induced edema in mice and rats, the aqueous fraction being distinctly more effective. When studied on prostaglandin E₂ (PGE₂), arachidonic acid, histamine, or serotonin (5HT)-induced edema in rats, Areca catechu and its aqueous fraction markedly repressed only the PGE₂ and arachidonic acid-induced inflammation. When studied for analgesic activity, the crude extract and its aqueous fraction produced a dose-dependent (10-100 mg/kg) inhibitory effect on formalin-induced nociception in mice and acetic acid-induced writhing in rats, similar to aspirin. In DPPH assay, Areca catechu and its aqueous fraction exhibited free radical scavenging activity with respective IC(50) values of 5.34 μg/ml (4.93-5.78, CI; 95%, n=5) and 7.28 μg/ml (6.04-7.95, n=4), like that of rutin with IC(50) value of 4.75 μg/ml (3.89-5.42, n=4). These results indicate the anti-inflammatory and analgesic effects of Areca catechu and provide a rationale for its medicinal use in inflammatory disorders.

IKKα and IKKβ Each Function to Regulate NF-κB Activation in the TNF-Induced/Canonical Pathway

BackgroundActivation of the transcription factor NF-κB by cytokines is rapid, mediated through the activation of the IKK complex with subsequent phosphorylation and degradation of the inhibitory IκB proteins. The IKK complex is comprised of two catalytic subunits, IKKα and IKKβ, and a regulatory protein known as NEMO. Using cells from mice that are genetically deficient in IKKβ or IKKα, or using a kinase inactive mutant of IKKβ, it has been proposed that IKKβ is critical for TNF-induced IκB phosphorylation/degradation through the canonical pathway while IKKα has been shown to be involved in the non-canonical pathway for NF-κB activation. These conclusions have led to a focus on development of IKKβ inhibitors for potential use in inflammatory disorders and cancer.MethodologyAnalysis of NF-κB activation in response to TNF in MEFs reveals that IKKβ is essential for efficient phosphorylation and subsequent degradation of IκBα, yet IKKα contributes to the NF-κB activation response in these cells as measured via DNA binding assays. In HeLa cells, both IKKα and IKKβ contribute to IκBα phosphorylation and NF-κB activation. A kinase inactive mutant of IKKβ, which has been used as evidence for the critical importance of IKKβ in TNF-induced signaling, blocks activation of NF-κB induced by IKKα, even in cells that are deficient in IKKβ.ConclusionsThese results demonstrate the importance of IKKα in canonical NF-κB activation, downstream of cytokine treatment of cells. The experiments suggest that IKKα will be a therapeutic target in inflammatory disorders.

Anti-inflammatory activities of Sieboldogenin from Smilax china Linn.: Experimental and computational studies

Ethnopharmacological relevance Smilax china Linn. is extensively used in traditional Chinese medicine (TCM) as well as in Pakistan for several medicinal purposes including their use in inflammatory disorders. Aim of the study The aims of the current study were to validate and assess the folk use of Smilax china Linn. on pharmacological grounds using the isolated compound at molecular, in vivo and computational levels. Materials and methods Seiboldogenin was isolated from ethyl acetate fraction of the plant crude extract. In vitro lipoxygenase and in vivo carrageenan-induced hind paw oedema models were used in experimental studies while molecular docking technique was used to conduct computational study. Results Sieboldogenin showed significant lipoxygenase inhibition (IC50: 38 μM). It also exhibited significant inhibition ( p < 0.05) of carrageenan-induced hind paw oedema at the doses of 10 and 50 mg/kg. Computational molecular docking showed its molecular interaction with important amino acid residues in the catalytic site of lipoxygenase, revealing its potential binding mode at molecular level. Conclusions Sieboldogenin seems to be a potential new anti-inflammatory compound responsible for anti-inflammatory activities of Smilax china Linn. Its in vitro and in vivo inflammatory activities are in good agreement with the folk medicinal use of Smilax china Linn. in inflammatory disorders.

Probiotics and Commensals Reverse TNF-α– and IFN-γ–Induced Dysfunction in Human Intestinal Epithelial Cells

Commensal bacteria are crucial for the development of the mucosal immune system. Probiotics are commensals with special characteristics and may protect mucosal surfaces against pathogens. Pathogens cause significant phenotypic alterations in infected epithelial cells, and probiotics reverse these deleterious responses. We hypothesized that probiotics and/or commensals may also reverse epithelial damage produced by cytokines. Human intestinal epithelial cells were exposed basolaterally to interferon (IFN)-gamma (10(3) U/mL) or tumor necrosis factor (TNF)-alpha (10 ng/mL) for up to 48 hours and assessed for ion transport, transepithelial resistance (TER), and epithelial permeability in the presence or absence of probiotics (Streptococcus thermophilus [ST] and Lactobacillus acidophilus [LA]), or the commensal, Bacteroides thetaiotaomicron (BT). Agonist-stimulated chloride secretion was inhibited by IFN-gamma, an effect prevented by ST/LA or BT. The ability of ST/LA or BT to restore Cl(-) secretion was blocked by inhibitors of p38 MAPK, ERK1, 2, and PI3K. The cystic fibrosis transmembrane conductance regulator (CFTR) and the NKCC1 cotransporter were down-regulated by IFN-gamma, and ST/LA pretreatment reversed this effect. Both TNF-alpha and IFN-gamma significantly reduced TER and increased epithelial permeability, effects prevented by ST/LA or BT. A Janus kinase (JAK) inhibitor synergistically potentiated effects of ST/LA or BT on TER and permeability, but p38, ERK1, 2, or PI3K inhibition did not. Finally, only probiotic-treated epithelial cells exposed to cytokines showed reduced activation of SOCS3 and STAT1,3. Deleterious effects of TNF-alpha and IFN-gamma on epithelial function are prevented by probiotic, and to a lesser extent, commensal pretreatment. These data extend the spectrum of effects of such bacteria on intestinal epithelial function and may justify their use in inflammatory disorders.