NF κB Research Articles

Novel Delivery Technologies: Triggering the Biopharmaceutical Potential of Boswellic Acids

Abstract: Boswellic acids are a series of pentacyclic triterpenes derived from the gum resin of Boswellia Genus, mostly from Boswellia serrata Roxb. (Burseraceae) tree commonly known as Indian Frankincense or salai guggul and traditionally used as an anti-inflammatory agent. It acts by inhibiting 5-Lipoxygenase, C3-convertase, cyclooxygenase, preferably COX-1, Human Leukocyte Elastase, NF- κB expression, Topoisomerase I and II and microsomal Prostaglandin E2 synthase-1. Boswellia species are reported to have various pharmacological potentials like anti-inflammatory, anti-cancer, antimicrobial, anti-arthritic, immunomodulatory activity, neuroprotective activity, and are also proved to be effective against ileitis, ulcerative colitis, hypolipidemic, hypertension and hepatotoxicity. Regardless of their multiple uses, pharmacokinetic studies of Boswellic acids revealed their poor oral bioavailability, high lipophilicity, and their degradation by the hepatic Phase I mechanism. With low solubility and poor bioavailability, different approaches have been applied to improve the poor pharmacokinetic profile of Boswellic acids. Designing and developing novel delivery systems for their enhanced permeability and improved bioavailability with better efficacy have been of great interest.

Empagliflozin inhibits neointimal hyperplasia through attenuating endothelial-to-mesenchymal transition via TAK-1/NF-κB pathway

ObjectiveTo investigate whether empagliflozin could prevent injury-induced vascular neointimal hyperplasia and to further explore its mechanism. MethodsMale C57BL/6J mice were divided into two groups with or without the empagliflozin treatment, and carotid ligation injury was performed to induce neointimal hyperplasia. The injured carotid arteries were collected for Western blotting (WB), histology and immunofluorescence analysis after four weeks. The inflammatory responses were analyzed by qRT-PCR to detect the inflammatory gene mRNA expression. To further explore its mechanism, HUVECs were treated with TGFβ-1 to induce EndMT followed by empagliflozin or vehicle treatment in vitro. A23187 (Calcimycin), an agonist of NF-κB signaling was used in the experiment. ResultsThe wall thickness and the neointima area was significantly reduced in the empagliflozin treatment group on day 28 after artery ligation. The Ki-67 positive cells were 28.33 ± 12.66% and 48.83 ± 10.41% in the empagliflozin-treated group and control group, respectively (P < 0.05). The mRNA expression levels of the inflammatory genes and inflammatory cells were decreased in the empagliflozin treatment group, as well as the MMP2 and MMP9. Meanwhile, empagliflozin can significantly reduce the migratory ability of inflammatory-treated HUVECs. The CD31 was increased in the TGFβ1+empagliflozin group, whereas the FSP-1, phosphorylation of TAK-1 (p-TAK-1) and phosphorylation of NF-κB (p- NF-κB) expression level were decreased, compared to the control group without empagliflozin treatment. However, the expression level of FSP-1 and p–NF–κB were reversed after co-treatment with A23187, whereas the p-TAK-1 expression level was without any significant difference. ConclusionEmpagliflozin inhibits the inflammation-induced EndMT via the TAK-1/NF-κB signaling pathway.

Norcantharidin alleviates cyclophosphamide induced immunosuppression and leukopenia in mice through NF κB pathway

Purpose: To investigate the effect of norcantharidin (NCTD) on cyclophosphamide (CY)-induced immunosuppression and leukopenia in mice.&#x0D; Methods: Cyclophosphamide (80 mg/kg/day) was intraperitoneally administered to mice from day 1 to 3 and day 9, and from day 4 to 10, NCTD (1, 2 and 4 mg/kg) was intraperitoneally administered. The counts of white blood cells (WBC), red blood cells (RBC) and platelets (PLT) in peripheral blood were determined, followed by the evaluation of mouse spleen and thymus indices. Histopathological examination was performed using hematoxylin-eosin (H &amp; E) staining. In addition, the serum levels of immunoglobulins and cytokines were determined using enzyme-linked immunosorbent assay (ELISA), while plenetic T lymphocyte subsets were assessed by flow cytometry. The expression levels of p65, p-p65, IκBα, and p-IκBα in spleen tissues were determined by immunoblotting.&#x0D; Results: Administration of NCTD (1, 2 and 4 mg/kg) increased blood cell counts, and alleviated injuries on splenocytes and thymocytes in CY-treated mice, accompanied by decline in spleen and thymus indices. NCTD ameliorated CY-induced immunosuppression by inhibiting the cytokines but increased the immunoglobulins. CD4+ and CD8+ T lymphocyte expressions were enhanced, especially at the high dose of NCTD (4 mg/kg). Besides, NCTD at high doses (2 and 4 mg/kg) significantly activated NF-κB pathway.&#x0D; Conclusion: Administration of NCTD alleviates CY-induced leukopenia and immunosuppression in mice through activation of NF-κB pathway, thus providing a lead for the potential treatment of chemotherapy-induced leukopenia and immunosuppression.

Physalin A alleviates inflammation and oxidative stress in mouse infantile pneumonia by inhibiting JAK/STAT3 and NF κB pathways

Purpose: To investigate the effect of physalin A on infantile pneumonia in mice.&#x0D; Methods: A mouse model of infantile pneumonia was established via intraperitoneal injection of lipopolysaccharide and verified using hematoxylin and eosin staining. Lipopolysaccharide-stimulated mice were then administered physalin A. The total protein content, number of cells, and levels of inflammatory-factor in bronchoalveolar lavage fluid (BALF) were determined. Inflammation and oxidative stress levels in lung tissues were determined by enzyme-linked immunosorbent assay (ELISA).&#x0D; Results: Lipopolysaccharide injection induced shrinking of pulmonary alveoli in mice, but physalin A administration ameliorated the histopathologic damage in lung tissues and significantly reduced the total protein content and number of cells in the BALF of lipopolysaccharide-stimulated mice (p &lt; 0.001). Moreover, physalin A also significantly down-regulated the levels of tumor necrosis factor-α, interleukin (IL)-6, IL-18, and IL-1β in BALF and lung tissues of lipopolysaccharide-treated mice (p &lt; 0.001). Physalin A attenuated lipopolysaccharide-induced increases in malondialdehyde and myeloperoxidase as well as decreases in superoxide dismutase and glutathione in mouse lung tissues. Additionally, physalin A reduced the levels of p-JAK1, p-STAT3, and p-p65 in lung tissues of lipopolysaccharide-treated mice.&#x0D; Conclusion: Physalin A exerts anti-inflammatory and anti-oxidant effect on lipopolysaccharide-induced lung injury in mice through the inactivation of JAK/STAT3 and NF-κB pathways. However, the effect of Physalin A on inflammation and oxidative stress in lipopolysaccharide-induced A549 cells will need to be investigated in further studies.&#x0D;

Protective effect of epalrestat on peripheral nerves in rats with diabetic peripheral neuropathy via NF κB pathway

Purpose: To study the protective effect of epalrestat on peripheral nerves in rats with diabetic peripheral neuropathy (DPN).&#x0D; Methods: A total of 36 Sprague-Dawley rats were randomly divided into normal, model, and epalrestat groups, each containing 12 rats. The morphology of the neurons was assessed using hematoxylin-eosin (H&amp;E) staining. The expressions of B-cell lymphoma-2 (Bcl-2), and Bcl-2 associated X protein (Bax) were determined via immunohistochemistry. The relative protein expressions of NF-κB and Caspase3 were determined via Western blotting, and apoptosis was determined using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay.&#x0D; Results: The morphology of the neurons was clear and intact in the normal group, disordered and destroyed in the model group, and was improved in the epalrestat group. The other two groups had a significantly higher positive expression of Bax and a significantly lower positive expression of Bcl-2 than the normal group (p &lt; 0.05), while the epalrestat group had a significantly lower positive expression of Bax, and a significantly higher positive expression of Bcl-2 than the model group (p &lt; 0.05). Furthermore, the protein expressions of NF-κB and Caspase3 were increased in other groups compared with normal group (p &lt; 0.05), while they declined in epalrestat group compared with model group (p &lt; 0.05). The apoptosis rate was significantly lower in epalrestat group than in model group (p &lt; 0.05).&#x0D; Conclusion: Epalrestat inhibits neuronal apoptosis by suppressing the NF-κB signaling pathway, thereby exerting a neuroprotective effect in DPN rats. Further studies would be required to validate the molecular mechanism.

The combination of Alisma and Atractylodes ameliorates cerebral ischaemia/reperfusion injury by negatively regulating astrocyte-derived exosomal miR-200a-3p/141-3p by targeting SIRT1

Ethnopharmacological relevanceThe combination of Alisma and Atractylodes (AA), a classical traditional Chinese herbal decoction, may protect against cerebral ischaemia/reperfusion injury (CIRI). However, the underlying mechanism has not been characterized. Intriguingly, exosomal microRNAs (miRNAs) have been recognized as vital factors in the pharmacology of Chinese herbal decoctions. Aim of the studyThe aim of the present study was to assess whether the neuroprotective effect of AA was dependent on the efficient transfer of miRNAs via exosomes in the brain. Materials and methodsBilateral common carotid artery ligation (BCAL) was used to induce transient global cerebral ischaemia/reperfusion (GCI/R) in C57BL/6 mice treated with/without AA. Neurological deficits were assessed with the modified neurological severity score (mNSS) and Morris water maze (MWM) test. Western blot (WB) analysis was used to detect the expression of sirtuin 1 (SIRT1) in the cerebral cortex. The inflammatory state was quantitatively evaluated by measuring the expression of phospho-Nuclear factor kappa B (p–NF–κB), Interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) using WB analysis and glial fibrillary acidic protein (GFAP) immunohistochemical staining. The protein expression of zonula occluden-1 (ZO-1), occludin, caudin-5 and CD31 was examined by immunohistochemical staining to determine blood‒brain barrier (BBB) permeability. Exosomes were extracted from the brain interstitial space by ultracentrifugation and identified by transmission electron microscopy (TEM), WB analysis and nanoparticle tracking analysis (NTA). The origin of exosomes was clarified by measuring the specific mRNAs within exosomes via Real Time Quantitative PCR (RT‒qPCR). Differential miRNAs in exosomes were identified using microarray screening and were validated by RT‒qPCR. Exosomes were labelled with fluorescent dye (PKH26) and incubated with bEnd.3 cells, the supernatant was collected, IL-1β/TNF-α expression was measured using enzyme-linked immunosorbent assay (ELISA), total RNA was extracted, and miR-200a-3p/141-3p expression was examined by RT‒qPCR. In addition, the levels of miR-200a-3p/141-3p in oxygen glucose deprivation/reoxygenation (OGD/R)-induced bEnd.3 cells were quantified. The direct interaction between miR-200a-3p/141-3p and the SIRT1 3′ untranslated region (3′UTR) was measured by determining SIRT1 expression in bEnd.3 cells transfected with the miR-200a-3p/141-3p mimic/inhibitor. ResultsSevere neurological deficits and memory loss caused by GCI/R in mice was markedly ameliorated by AA treatment, particularly in the AA medium-dose group. Moreover, AA-treated GCI/R-induced mice showed significant increases in SIRT1, ZO-1, occludin, caudin-5, and CD31 expression levels and decreases in p–NF–κB, IL-1β, TNF-α, and GFAP expression levels compared with those in untreated GCI/R-induced mice. Furthermore, we found that miR-200a-3p/141-3p was enriched in astrocyte-derived exosomes from GCI/R-induced mice and could be inhibited by treatment with a medium dose of AA. The exosomes mediated the transfer of miR-200a-3p/141-3p into bEnd.3 cells, promoted IL-1β and TNF-α release and downregulated the expression of SIRT1. No significant changes in the levels of miR-200a-3p/141-3p were observed in OGD/R-induced bEnd.3 cells. The miR-200a-3p/141-3p mimic/inhibitor decreased/increased SIRT1 expression in bEnd.3 cells, respectively. ConclusionOur findings demonstrated that AA attenuated inflammation-mediated CIRI by inhibiting astrocyte-derived exosomal miR-200a-3p/141-3p by targeting the SIRT1 gene, which provided further evidence and identified a novel regulatory mechanism for the neuroprotective effects of AA.

Systems pharmacology reveals the mechanism of Astragaloside IV in improving immune activity on cyclophosphamide-induced immunosuppressed mice

Ethnopharmacological relevanceMyelosuppression, also known as bone marrow suppression (BMS), is a pathological phenomenon of the decrease in the production of blood cells and further lead to immune homeostasis disorder. Astragalus mongholicus Bunge (AM, checked with The World Flora Online, http://www.worldfloraonline.org, updated on January 30, 2023) is a traditional Chinese medicine with efficacy of tonifying Qi and strengthening body immunity in thousands of years of clinical practice in China. Astragaloside IV (AS-IV) is a major active ingredient of AM, which plays an important role in regulating immune system through different ways. Aim of the studyThis study was aimed to investigate the protective effect and mechanism of AS-IV on macrophages in vitro and cyclophosphamide (CTX)-induced immunosuppressive mice in vivo, and to provide experimental basis for the prevention and treatment of AS-IV in myelosuppression. Materials and methodsBased on network pharmacology and molecular docking technology, the core targets and signaling pathways of saponins of AM against myelosuppression were screened. And then, the immunoregulatory effect of AS-IV on RAW264.7 cells was investigated by cellular immune activity and cellular secretion analysis in vitro. In this way, the effects of AS-IV on the main potential targets of HIF-1α/NF-κB signaling pathway were analyzed by qRT-PCR and Western blot methods. Furthermore, comprehensive analysis of the effects of AS-IV against CTX-induced mice were conducted on the basis of immune organs indices analysis, histopathological analysis, hematological analysis, natural killer cell activity analysis and spleen lymphocyte transformation activity analysis. In order to further verify the relationship between active ingredients and action targets, drug inhibitor experiments were finally conducted. ResultsAS-IV, as a potential anti-myelosuppressive compound, was screened by systematic pharmacological methods to act on target genes including HIF1A and RELA together with the HIF-1α/NF-κB signaling pathway. Further studies by molecular docking technology showed that AS-IV had good binding activity with HIF1A, RELA, TNF, IL6, IL1B and other core targets. Besides, cellular and animal experiments validation results showed that AS-IV could enhance the migration and phagocytosis of RAW264.7 cells, and protect the immune organs such as spleen and thymus together with bone tissues from damage. By this means, immune cell function including spleen natural killer cell and lymphocyte transformation activity were also enhanced. In addition, white blood cells, red blood cells, hemoglobin, platelets and bone marrow cells were also significantly improved in the suppressed bone marrow microenvironment (BMM). In kinetic experiments, the secretion of cytokines such as TNF-α, IL-6 and IL-1β were increased, and IL-10, TGF-β1 were decreased. The key regulatory proteins such as HIF-1α, NF-κB, PHD3 in HIF-1α/NF-κB signaling pathway were also regulated in the results of upregulated expression of HIF-1α, p–NF–κB p65 and PHD3 at the protein or mRNA level. Finally, the inhibition experiment results suggested that AS-IV could significantly improve protein response in immunity and inflammation such as HIF-1α, NF-κB and PHD3. ConclusionAS-IV could significantly relieve CTX-induced immunosuppressive and might improve the immune activity of macrophages by activating HIF-1α/NF-κB signaling pathway, and provide a reliable basis for the clinical application of AS-IV as a potentially valuable regulator of BMM.

Gomisin A inhibits hypoxia/reoxygenation induced myocardial cell injury by modulating TLR4 NF κB pathway

Purpose: To determine the impact of Gomisin A (GomA) on myocardial cell damage caused by hypoxia/reoxygenation (H/R). Methods: Various methods, including MTT, western blot and flow cytometry were used to to assess the viability of H9c2 cardiomyocytes and cell apoptosis. Expression levels of various enzymes and cytokines, including superoxide dismutase (SOD), malondialdehyde (MDA), catalase (CAT), tumor necrosis factor-alpha (TNF-α), interleukin (IL)-1β and IL-6 were measured by enzyme-linked immunosorbent assay (ELISA). Western blot was used to evaluate the expression levels of inducible nitric oxide synthase (iNOS), cyclooxygenase 2 (COX-2) and Toll-like receptor 4 (TLR4), as well as the phosphorylation of p65 and nuclear factor kappa B (NF-κB) inhibitor alpha (IκBα). Results: Low concentrations of GomA had no toxic effect on H9c2 cells. In H/R-stressed H9c2 cells, GomA increased cell viability and reduced cell apoptosis, suggesting that GomA inhibited H/R-induced cell apoptosis. Additionally, GomA alleviated H/R-induced oxidative stress and inflammation by increasing the expression of SOD and CAT, while decreasing the expression of MDA, iNOS, COX-2, TNF-α, IL-1β, and IL-6. GomA also suppressed the H/R-induced TLR4–NF-κB pathway by reducing the expression of TLR4 and the phosphorylation of p65 and IκBα. Conclusion: These results indicate that GomA is a potential candidate for the treatment of myocardial ischemia/reperfusion (MI/R) injury by increasing cell viability, reducing cell apoptosis, alleviating oxidative stress, and reducing inflammation by inhibiting TLR4–NF-κB pathway.

Exploration of bioactive compounds from Mangifera indica (Mango) as probable inhibitors of thymidylate synthase and nuclear factor kappa-B (NF-Κb) in colorectal cancer management

Abstract This research is aimed at investigating the anti-colorectal cancer activities of phytochemicals from Mangifera indica (Mango) via the inhibition of thymidylate synthase (TS) and Nuclear Factor kappa B (NF–κB) using computational chemistry tools. Ligands (141 phytochemicals previously isolated from mangoes) and reference drugs (Raltitrexed and Emetine), the drug inhibitors of TS and NF–κB, respectively) were subjected to screening via ADMET profiling, drug-likeness analysis, oral bioavailability, PASS profile, and molecular interactions. Ligands that passed the previously mentioned screening were docked in duplicate against the target receptors (TS and NF–κB) using PyRx software. The mean values were calculated to obtain suitable docking scores. The analysis showed that TS was strongly inhibited by Friedelan-3beta-Ol with its lower binding energy of −9.0 kcal/mol more than Raltitrexed with a binding energy of −8.7 kcal/mol. NF–κB was also inhibited by Friedelan-3beta-Ol and Friedelin with binding energies of −8.0 and −8.1 kcal/mol, respectively, more than Emetine with a binding energy of −6.4 kcal/mol. These two phytochemicals performed much better than the standard drugs, thus selected as the best hits compounds because of their ADMET profile, drug-likeness properties, bioactivity, oral bioavailability, PASS prediction, binding affinities, and their interactions with the amino acids in the active sites of the receptors. Therefore, further studies are necessary for the validation of these claims toward the development of new effective and safer anti-colorectal cancer drugs.

Identification of potential inhibitors of thymidylate synthase (TS) (PDB ID: 6QXH) and nuclear factor kappa-B (NF–κB) (PDB ID: 1A3Q) from Capsicum annuum (bell pepper) towards the development of new therapeutic drugs against colorectal cancer (CRC)

Abstract Colorectal cancer is the third most deadly cancer globally. Drug resistance and attendant side effects make the available standard anti-colorectal cancer drugs against target receptors inefficient. Phytochemicals from medicinal plants are safer, cheaper, effective, and heal diseases from the cellular level. This study is aimed at identifying potential inhibitors of thymidylate synthase (TS) and nuclear factor kappa-B (NF–κB) target receptors from Capsicum annuum towards the development of new therapeutic drugs against colorectal cancer via in silico approach. One hundred and fifty (150) ligands previously reported from Capsicum annuum were downloaded from the PubChem database and were subjected to chemo-informatics analyses such as ADMET, drug-likeness, oral bioavailability, bioactivity, and PASS prediction to ascertain their therapeutic and safety profile before docking. The ligands that passed the analyses were docked against TS and NF–κB in duplicate using a creditable docking tool (PyRx). Raltitrexed and emetine were used as the standard drug inhibitors for TS and NF–κB, respectively. The results obtained from this study showed that feruloyl-beta-D-glucose (8.45 kcal/mol), 5-O-caffeoylquinic acid (−8.40 kcal/mol), 5-O-caffeoylquinic acid methyl ester (−7.89 kcal/mol), feruloyl hexoside (−7.40 kcal/mol), O-glucopyranoside (−7.55 kcal/mol), and quercetin (−7.00 kcal/mol) shared the same binding pocket with TS while feruloyl-beta-D-glucose (−7.00 kcal/mol), chlorogenic acid (−6.90 kcal/mol), 5-O-caffeoylquinic acid (−6.90 kcal/mol) and feruloyl hexoside (−6.50 kcal/mol) shared the same pocket with NF–κB. These compounds were selected as best hits due to their excellent inhibitory efficiency and chemoinformatic profiles. Thus, the compounds may function as prospective lead compounds for developing a new anti-colorectal cancer drug.

Hypaphorine ameliorates lipid accumulation and inflammation in a cellular model of non alcoholic fatty liver by regulating p38/JNK and NF κB signaling pathways

Purpose: To investigate the therapeutic effect and underlying mechanism of hypaphorine in a cellular model of non-alcoholic fatty liver disease (NAFLD).Methods: Palmitic acid (PA) was used to induce a NAFLD phenotype in hepatocytes. Cell viability and apoptosis were evaluated by CCK-8 and flow cytometry assays. Inflammatory response was measured by enzyme-linked immunosorbent assay (ELISA). The effect of hypaphorine on lipid accumulation was evaluated using Oil Red O staining and triglyceride kits. Activation of p38/c–Jun N-terminal kinase (JNK) and NF-κB pathways were analyzed by immunoblot assay.Results: Hypaphorine significantly improved cell viability (p &lt; 0.01), suppressed inflammatory response (p &lt; 0.01), and reduced lipid accumulation (p &lt; 0.01) in PA-treated hepatocytes. Hypaphorine ameliorated lipid accumulation and inflammation in PA-treated hepatocytes by targeting p38/JNK and NF-κB pathways.Conclusion: Hypaphorine may serve as a therapeutic target in NAFLD. However, in vivo studies to validate this finding are required.

Transcriptome-wide m6A modification mediates cardiotoxicity in mice after chronic exposure to microplastics

The widespread presence of microplastics (MPs) has garnered attention owing to their possible adverse effects. However, the cardiotoxicity of MPs and their underlying mechanism remains unclear. N6-methyladenosine (m6A) modification contributes to post-transcriptional modulation of gene expression, but whether this modification is relevant to MP-induced cardiotoxicity is unknown. First, we detected damage to the myocardial tissue among MP-exposed and -unexposed (control) mice by staining them with hematoxylin and eosin, Masson trichrome, and Oil Red O. Then, we comprehensively measured the transcriptome-wide m6A methylome and m6A-altered genes using high-throughput sequencing assays, such as methylated RNA immunoprecipitation sequencing and RNA sequencing. Our data indicated MP-induced myocardial inflammatory injury and histological alterations in vivo, evidenced by the severity of cardiac fibrosis and increased lipid accumulation. We found 878 increased and 316 decreased methylation peaks mostly distributed in the 3′ UTR among the MP-exposed group compared with the control group. We found 779 upregulated and 340 downregulated genes in the MP-exposed group. In addition, conjoint analysis of results from the two high-throughput sequencings showed that 109 and 11 hypermethylated genes were upregulated and downregulated, respectively; 12 and 21 hypomethylated genes were upregulated and downregulated, respectively. Results of the cross-link analysis showed that several potential signals, such as ECM–receptor interactions, cell adhesion molecules, cytokine–cytokine receptor interactions, and NF–κB signaling, contributed to MP-induced cardiotoxicity. Our findings indicated that m6A modifications of genes were involved in MP-induced cardiotoxicity and reported new information regarding the chronic cardiotoxicity caused by MP exposure in mice.

Caulerpa lentillifera improves ethanol-induced liver injury and modulates the gut microbiota in rats

Caulerpa lentillifera (CL), also called sea grape, is a type of edible green alga which was reported to have antioxidative and immunomodulatory potential. This study aimed to investigate the hepatoprotective effects of CL in a rat model of chronic ethanol exposure. Wistar rats were assigned to four groups and supplied with an isocaloric control liquid diet (group C), an ethanol liquid diet (group E), a control liquid diet supplemented with 5% CL (group CC), or an ethanol liquid diet supplemented with 5% CL (group EC) for a 12-week experimental period. Ethanol feeding induced steatosis, inflammation, and changes in the gut microbiota by the end of the study, whereas CL supplementation significantly improved liver injuries and decreased circulatory endotoxin levels. Moreover, we also found that CL reversed ethanol-induced elevation of hepatic toll-like receptor 4 (TLR4), MyD88 protein expression, the phosphorylated-nuclear factor (NF)-κB-to–NF–κB ratio, and proinflammatory cytokine concentrations. Additionally, CL also increased the abundance of Akkermansia and tight junction proteins and diminished the Firmicutes-to-Bacteroidetes ratio. Dietary CL inhibited the progression of alcoholic liver disease, and some of the possible mechanisms may be strengthening the intestinal barrier function, alleviating dysbiosis, and modulating the TLR4 pathway.

Tryptophan metabolism can modulate immunologic tolerance in primitive vertebrate lamprey via IDO-kynurenine-AHR pathway

Tryptophan is mainly degraded through kynurenine pathway (KP) in vertebrates which is closely related to the nerve and depression, while the studies on immunity is still limited. This study aims to explore the functions of tryptophan in the innate immunity of primitive vertebrate lamprey. MTT (3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide) assay showed that tryptophan had no obvious effect on cell viability. Tryptophan was transported into leukocytes and degraded via the KP after tryptophan supplement. Tryptophan treatment (T1x and T2x) failed to alter the total antioxidant capacity regardless of stimulation and exposure time. Real-time quantitative PCR and western blotting results revealed that tryptophan was not only able to reduce the expression of pro-inflammatory factors Lj-TNF-α, Lj-IL1β and Lj–NF–κB, but also to upregulate the expression of anti-inflammatory factor Lj-TGF-β independent of stimulation and time. In addition, tryptophan can exert immune tolerance function by inhibiting TLR-MyD88 and promoting (Indoleamine 2, 3-Dioxygenase) IDO-kynurenine-AHR (aryl hydrocarbon receptor) pathways. This study provides a new understanding for tryptophan-kynurenine metabolism and mechanism of immune tolerance function in primitive vertebrate lamprey.

Alternanthera brasiliana L. extract alleviates carbon tetrachloride-induced liver injury and fibrotic changes in mice: Role of matrix metalloproteinases and TGF-β/Smad axis

Ethnopharmacological relevanceAlternanthera brasiliana L. is a flowering plant belonging to the family Amaranthaceae and is popularly known as “penicillin”. It is used in folk medicine to treat infections, coughs, wound healing, and inflammatory diseases.Aim of the study: We investigated the effect of Alternanthera brasiliana L. leaves hydroalcoholic extract (AB) against oxidative stress, inflammation, and fibrotic changes in an experimental model of carbon tetrachloride (CCl4)-induced liver injury and fibrosis in mice. Materials and methodsThirty-six male Balb/C mice were randomized into five groups: normal control, AB control, CCl4 control, CCl4 + AB-200 mg/kg, and CCl4 + AB-400 mg/kg. In mice, liver injury was induced by intraperitoneal injection of CCl4 (20% in corn oil, 5 ml/kg body weight) thrice a week for six consecutive weeks. AB extract at two doses (200 mg/kg and 400 mg/kg body weight) was administered orally for six consecutive weeks. Liver injury-related serum markers (ALT, AST, ALP), antioxidants (GSH, GST, SOD, and vitamin C), pro-inflammatory cytokines including tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-18, ultrasonographic and histological alterations, proteins of matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinase-1 (TIMP-1), nuclear factor-κB (p65) (NF-κB), nod-like receptor protein 3 (NLRP3), and TGF-β/Smad signaling were accessed. LC-Q-TOF-MS/MS analysis of AB was performed. ResultsAB treatment significantly decreased the CCl4-induced rise in serum ALT, AST, and ALP activities and improved the histological alterations. Compared with the CCl4-treated group, treatment with AB significantly restored the hepatic antioxidants and reduced the pro-inflammatory cytokines in the liver. The antioxidant activity of AB may be attributed to its terpenoid constituents, which was confirmed by LC-Q-TOF-MS/MS analysis. The CCl4-induced rise in expression of MMP-2 and MMP-9 and decrease in TIMP-1 were markedly restored in the AB-treated groups. Further findings revealed a significant reduction in the protein levels of phospho–NF–κB (p65), NLRP3, TGF-β, pSmad2/3, collagen I, and α-smooth muscle actin (α-SMA) in the AB treatment groups. ConclusionsThe hepatoprotective effect of AB may be attributed to the high content of terpenoid compounds and alleviates liver injury and associated fibrotic changes through modulating MMPs, NF-κB (p65), and the TGF-β/Smad axis.

Honokiol inhibits interleukin-induced angiogenesis in the NSCLC microenvironment through the NF-κB signaling pathway

Tumor angiogenesis, which may be affected by microenvironmental inflammation and promotes tumor development and metastasis, is one of the key reasons contributing to increased mortality. The goal of this study is to investigate how lignin analogs, specifically honokiol (HNK), block angiogenesis induced by the inflammatory milieu of lung cancer. The human lung cancer cell lines A549 and H460 were treated with HNK. Interleukin-1 was employed to mimic an inflammatory tumor microenvironment. Findings demonstrated that HNK drastically decreased the cell viability of A549 and H460 cells. In A549 and H460 cells, HNK also reduced the production of vascular endothelial growth factor (VEGF), the most important marker of tumor angiogenesis. Signal pathway studies revealed that HNK blocked the NF-κB signaling pathway. This effect, in turn, prevented the expression of VEGF by inhibiting the NF-κB signaling pathway. Human umbilical vein endothelial cells (HUVECs) from A549-conditioned medium cultures were subjected to HNK treatment, which decreased tubulogenesis, horizontal and vertical migration, and cell proliferation in HUVECs. Overall, HNK inhibited the NF–κB pathway. This effect resulted in the downregulation of VEGF, thus reducing the viability and angiogenesis of human lung cancer cell lines. In A549 cell xenografts, HNK decreased VEGF expression, tumor angiogenesis, and tumor development. Our research shows that HNK is a potential antiangiogenic molecule for the treatment of lung cancer.

Ophthalmic Manifestations of ROSAH (Retinal Dystrophy, Optic Nerve Edema, Splenomegaly, Anhidrosis, and Headache) Syndrome, an Inherited NF κB–Mediated Autoinflammatory Disease with Retinal Dystrophy

We aimed to characterize the ocular phenotype of patients with ROSAH (retinal dystrophy, optic nerve edema, splenomegaly, anhidrosis, and headache) syndrome and their response to therapy. Single-center observational case study. Eleven patients with a diagnosis of ROSAH syndrome and mutation in ALPK1 were included. Patients with molecularly confirmed ROSAH syndrome underwent ophthalmic evaluation, including visual acuity testing, slit-lamp and dilated examinations, color fundus and autofluorescence imaging, fluorescein angiography, OCT, and electrophysiologic testing. Visual acuity, electrophysiology, fluorescein angiography, and OCT findings. Eleven individuals (6 female and 5 male patients) from 7 families ranging in age from 7.3 to 60.2 years at the time of the initial evaluation were included in this study. Seven patients were followed up for a mean of 2.6 years (range, 0.33-5.0 years). Best-corrected visual acuity at baseline ranged from 20/16 to no light perception. Variable signs or sequelae of intraocular inflammation were observed in 9 patients, including keratic precipitates, band keratopathy, trace to 2+ anterior chamber cells, cystoid macular edema, and retinal vasculitis on fluorescein angiography. Ten patients were observed to show optic disc elevation and demonstrated peripapillary thickening on OCT. Seven patients showed retinal degeneration consistent with a cone-rod dystrophy, with atrophy tending to involve the posterior pole and extending peripherally. One patient with normal electroretinography findings and visual evoked potential was found to have decreased Arden ratio on electro-oculography. Leveraging insights from the largest single-center ROSAH cohort described to date, this study identified 3 main factors as contributing to changes in visual function of patients with ROSAH syndrome: optic nerve involvement; intraocular inflammation, including cystoid macular edema; and retinal degeneration. More work is needed to determine how to arrest the progressive vision loss associated with ROSAH syndrome. Proprietary or commercial disclosure may be found after the references.

Xuanfei Baidu Formula attenuates LPS-induced acute lung injury by inhibiting the NF-κB signaling pathway

Ethnopharmacological relevanceAcute lung injury (ALI) is a common manifestation of COVID-19. Xuanfei Baidu Formula(XFBD) is used in China to treat mild or common damp-toxin obstructive pulmonary syndrome in COVID-19 patients. However, the active ingredients of XFBD have not been extensively studied, and its mechanism of action in the treatment of ALI is not well understood. Aim of the studyThe purpose of this study was to investigate the mechanism of action of XFBD in treating ALI in rats, by evaluating its active components. Materials and methodsFirstly, the chemical composition of XFBD was identified using ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry. The potential targets of XFBD for ALI treatment were predicted using network pharmacological analysis. Finally, the molecular mechanism of XFBD was validated using a RAW264.7 cell inflammation model and a mouse ALI model. ResultsA total of 113 compounds were identified in XFBD. Network pharmacology revealed 34 hub targets between the 113 compounds and ALI. The results of Kyoto Encyclopedia of Genes and Genomes and gene ontology analyses indicated that the NF-κB signaling pathway was the main pathway for XFBD in the treatment of ALI. We found that XFBD reduced proinflammatory factor levels in LPS-induced cellular models. By examining the lung wet/dry weight ratio and pathological sections in vivo, XFBD was found that XFBD could alleviate ALI. Immunohistochemistry results showed that XFBD inhibited ALI-induced increases in p-IKK, p–NF–κB p65, and iNOS proteins. In vitro experiments demonstrated that XFBD inhibited LPS-induced activation of the NF-κB pathway. ConclusionThis study identified the potential practical components of XFBD, combined with network pharmacology and experimental validation to demonstrate that XFBD can alleviate lung injury caused by ALI by inhibiting the NF-κB signaling pathway.

Ningmitai capsules have anti-inflammatory and pain-relieving effects in the chronic prostatitis/chronic pelvic pain syndrome mouse model through systemic immunity.

Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) seriously affects the physical and mental health of approximately 90% of males. Due to its complex and unclear etiology, the treatment methods that are currently available for chronic prostatitis/chronic pelvic pain syndrome are controversial, and their efficacy is unsatisfactory. At present, most researchers believe that this kind of prostatitis is caused by autoimmune inflammation. Chinese herbs, which are the essence of traditional Chinese medicine (TCM), are emerging treatment options for inflammation and immune diseases. In this experiment, we investigated the effect of Ningmitai capsules (a kind of traditional Chinese medicine widely used to treat lower urinary tract inflammation and pain in males) on chronic prostatitis/chronic pelvic pain syndrome in a non-obese diabetes-experimental autoimmune prostatitis (NOD-EAP) mouse model. First, by using bioinformatics analysis of data from the Encyclopedia of Traditional Chinese Medicine (ETCM) database, we found that quercetin, which is one of the main components of Ningmitai capsules, could reduce the secretion of CCL2 by inhibiting the MAPK pathway. In animal experiments, it was found that after Ningmitai treatment, the inflammation in mouse prostates was alleviated, the expression of CCL2, which is related to pain, and MAPK pathway components were downregulated, and the activation of the inflammatory NF–κB and STAT3 pathways was reduced. Pelvic pain and inflammation were relieved in mice with EAP. Due to the presence of the blood–prostate barrier, the drug may not completely reach the prostate directly and take effect locally. However, we found that after Ningmitai treatment, the proportions of proinflammatory CD11b+Ly6Chigh immune cells in the spleen, bloodstream (systemic immunity), and prostate (local immunity) were reduced. The infiltration of CD11b+ immune cells into the spleen and prostate was decreased. These findings suggested that Ningmitai can treat chronic prostatitis/chronic pelvic pain syndrome by affecting systemic and local immunities through the CCL2–MAPK pathway.

Patchouli alcohol suppresses inflammation and autophagy, and reduces osteoclast formation in periodontitis via regulation of ERK and NF κB signaling pathways

Purpose: To evaluate the effect of patchouli alcohol (PA) on periodontitis and reveal its possible mechanism.&#x0D; Methods: Primary human periodontal membrane cells (PDLC) were isolated and treated with lipopolysaccharide (LPS) to establish a periodontitis model. The MTT, flow cytometry and immunofluorescence assay were used to determine the effect of PA on cell viability and apoptosis in PDLC exposed to LPS. The inflammation status and osteoclast formation were measured using enzyme-linked immunosorbent assay (ELISA) and TRAP staining. Immunoblot assays were performed to analyze the effect of LPS on ERK and NF-κB pathways.&#x0D; Results: Patchouli alcohol (PA) improved LPS-induced PDLC growth arrest (p &lt; 0.001). In addition, PA relieved LPS - induced autophagy of PDLCs (p &lt; 0.001). Further data confirmed that PA inhibited lPS-induced inflammation of PDLCs. In addition, PA reduced osteoclast formation, and suppressed inflammation and autophagy, and also reduced osteoclast formation by regulating ERK and NF-κB pathways.&#x0D; Conclusion: Patchouli alcohol inhibits inflammation and autophagy, and decreases osteoclast formation in periodontitis via the regulation of ERK and NF-κB signaling pathways. Thus, this compound has potentials for development for the treatment of periodontitis; however, in vivo studies should first be undertaken.