scholarly journals Anti-Inflammatory Potential of Hexane Extract of Mud Lobster (Thalassina anomala) in Lipopolysaccharide-Stimulated RAW 264.7 Macrophages

2021 ◽  
Vol 32 (1) ◽  
pp. 143-160
Author(s):  
Nur Nadiah Zakaria ◽  
◽  
Masnindah Malahubban ◽  
Sharida Fakurazi ◽  
Sie Chuong Wong ◽  
...  

Mud lobsters are crustaceans from the genus Thalassina which are lesser known and seldom seen but are nevertheless an important organism to the mangrove ecosystem. In Malaysia and Thailand, mud lobsters are eaten by locals as treatment for asthma. It is traditionally believed that they are effective in reducing the number of asthma attacks and severity of asthma symptoms. However, the therapeutic potential of mud lobster extract remains unclear and has not been fully elucidated or reported in any scientific study. The objectives of this study are to investigate the anti-inflammatory potential of mud lobster, Thalassina anomala extracts (hexane, chloroform and methanol) in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages, and to identify the potential bioactive compounds involved. An MTT assay was performed to determine the cytotoxicity of the T. anomala extracts on RAW 264.7 macrophages. Nitrite quantification assay and enzyme-linked immunosorbent assay (ELISA) were conducted to investigate the ability of the T. anomala extracts to suppress the secretion and expression of nitric oxide (NO), Prostaglandin E2 (PGE2) and proinflammatory cytokines (TNF-α, IL-6 and IL-1β) in LPS-stimulated macrophages. GC-MS analysis was done to identify putative metabolites. The hexane extract of T. anomala showed anti-inflammatory activity by significantly inhibiting the LPS-induced production of NO, PGE2, interleukin- (IL-) 6, IL-1β and tumour necrosis factor-alpha (TNF-α) in a concentration-dependent manner. Hexane extract treatment with 100 μg/mL has decreased the NO secretion into 37 μM. Meanwhile, hexane extract at concentration of 100 μg/mL able to significantly suppressed PGE2,TNF-α, IL-6 and IL-1β production into 2015 pg/mL, 2406 pg/mL, 460 pg/mL and 9.6 pg/mL, respectively. GC-MS analysis of the hexane extract revealed the presence of 19 putative compounds. The identified compounds were reported to have anti-inflammatory, antioxidant and antibacterial activities. These results suggest that the hexane extract of T. anomala potentially has anti-inflammatory properties and concentration dependently suppressed NO, PGE2 and proinflammatory cytokines’ production in LPS-stimulated macrophages. The findings provide a rational basis of the traditional use of mud lobster for inflammation-associated ailments.

2015 ◽  
Vol 2015 ◽  
pp. 1-11 ◽  
Author(s):  
Woan Sean Tan ◽  
Palanisamy Arulselvan ◽  
Govindarajan Karthivashan ◽  
Sharida Fakurazi

Aim of Study.Moringa oleiferaLam. (M. oleifera) possess highest concentration of antioxidant bioactive compounds and is anticipated to be used as an alternative medicine for inflammation. In the present study, we investigated the anti-inflammatory activity of 80% hydroethanolic extract ofM. oleiferaflower on proinflammatory mediators and cytokines produced in lipopolysaccharide- (LPS-) induced RAW 264.7 macrophages.Materials and Methods. Cell cytotoxicity was conducted by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Nitric oxide (NO) production was quantified through Griess reaction while proinflammatory cytokines and other key inflammatory markers were assessed through enzyme-linked immunosorbent assay (ELISA) and immunoblotting.Results. Hydroethanolic extract ofM. oleiferaflower significantly suppressed the secretion and expression of NO, prostaglandin E2(PGE2), interleukin- (IL-) 6, IL-1β, tumor necrosis factor-alpha (TNF-α), nuclear factor-kappa B (NF-κB), inducible NO synthase (iNOS), and cyclooxygenase-2 (COX-2). However, it significantly increased the production of IL-10 and IκB-α(inhibitor ofκB) in a concentration dependent manner (100 μg/mL and 200 μg/mL).Conclusion. These results suggest that 80% hydroethanolic extract ofM. oleiferaflower has anti-inflammatory action related to its inhibition of NO, PGE2, proinflammatory cytokines, and inflammatory mediator’s production in LPS-stimulated macrophages through preventing degradation of IκB-αin NF-κB signaling pathway.


Antioxidants ◽  
2018 ◽  
Vol 7 (12) ◽  
pp. 184 ◽  
Author(s):  
Fatiha Brahmi ◽  
Thomas Nury ◽  
Meryam Debbabi ◽  
Samia Hadj-Ahmed ◽  
Amira Zarrouk ◽  
...  

The present study consisted in evaluating the antioxidant, anti-inflammatory and cytoprotective properties of ethanolic extracts from three mint species (Mentha spicata L. (MS), Mentha pulegium L. (MP) and Mentha rotundifolia (L.) Huds (MR)) with biochemical methods on murine RAW 264.7 macrophages (a transformed macrophage cell line isolated from ascites of BALB/c mice infected by the Abelson leukemia virus). The total phenolic, flavonoid and carotenoid contents were determined with spectrophotometric methods. The antioxidant activities were quantified with the Kit Radicaux Libres (KRLTM), the ferric reducing antioxidant power (FRAP) and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays. The MS extract showed the highest total phenolic content, and the highest antioxidant capacity, while the MR extract showed the lowest total phenolic content and the lowest antioxidant capacity. The cytoprotective and anti-inflammatory activities of the extracts were quantified on murine RAW 264.7 macrophages treated with 7-ketocholesterol (7KC; 20 µg/mL: 50 µM) associated or not for 24 h and 48 h with ethanolic mint extracts used at different concentrations (25, 50, 100, 200 and 400 µg/mL). Under treatment with 7KC, an important inhibition of cell growth was revealed with the crystal violet test. This side effect was strongly attenuated in a dose dependent manner with the different ethanolic mint extracts, mainly at 48 h. The most important cytoprotective effect was observed with the MS extract. In addition, the effects of ethanolic mint extracts on cytokine secretion (Interleukin (IL)-6, IL-10, Monocyte Chemoattractant Protein (MCP)-1, Interferon (IFN)-ϒ, Tumor necrosis factor (TNF)-α) were determined at 24 h on lipopolysaccharide (LPS, 0.2 µg/mL)-, 7KC (20 µg/mL)- and (7KC + LPS)-treated RAW 264.7 cells. Complex effects of mint extracts were observed on cytokine secretion. However, comparatively to LPS-treated cells, all the extracts strongly reduce IL-6 secretion and two of them (MP and MR) also decrease MCP-1 and TNF-α secretion. However, no anti-inflammatory effects were observed on 7KC- and (7KC + LPS)-treated cells. Altogether, these data bring new evidences on the potential benefits (especially antioxidant and cytoprotective properties) of Algerian mint on human health.


2020 ◽  
Author(s):  
Anis Ben Hsouna ◽  
Rania Ben Saad ◽  
Wissal Dhifi ◽  
Marwa Khaled ◽  
Wissem Mnif ◽  
...  

Abstract Background: Lipid transfer proteins (LTP) are members of the family of pathogenesis-related proteins (PR-14) that play a key role in plant defense mechanisms.Methods: In this study, a novel gene TdLTP4 encoding an antifungal protein from wheat (cv. Om Rabiaa) was subcloned, overexpressed in Escherichia coli BL-21 (DE3) and enriched using ammonium sulfate fractionation. The TdLTP4 fusion protein was then tested against a panel of pathogens, food-borne and spoilage bacteria and fungi in order to evaluate the antimicrobial properties. Our protein was applied to 0.5 µg/mL LPS-induced RAW 264.7 macrophages in vitro at different concentrations (5, 10, 20, 50 and 100 µg/ml). Levels of nitric oxide (NO), pro-inflammatory cytokines interleukin (IL)-1β (IL-1 β), interleukin (IL)-6 (IL-6), tumor necrosis factor (TNF-α) and anti-inflammatory cytokine IL-10 in the supernatant fraction were measured using enzyme-linked immunosorbent assay (ELISA). Expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) were detected via Western blot. Results: The inhibition zones and minimal inhibitory concentration (MIC) values of bacterial strains were in the range of 14-26 mm and 62.5-250 µg/mL, respectively. Moreover, a remarkable activity against several fungal strains was revealed. TdLTP4 (5–100 µg/mL) decreased the production of NO (IC50= 4.32 µg/mL), IL-6 (IC50= 11.52 µg/mL), IL-1β (IC50= 7.87 µg/mL) and TNF-α (IC50 =8.66 µg/mL) by lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Our protein could modulate the macrophages inflammatory mode by causing reduction in iNOS and COX2.Conclusion: According to these findings, LTP fusion protein could be used as natural anti-inflammatory and antimicrobial agent in food preservation and human health.


2021 ◽  
Vol 19 (3) ◽  
pp. 355-363
Author(s):  
Jung-Wook Kang ◽  
In-Chul Lee

Purpose: This study aimed to investigate the effects of the Cassia obtusifolia L. seed extract (CSE) on particulate matter (PM)-induced skin.Methods: The effects of CSE on cell viability were evaluated using a skin cell line. To determine the anti-inflammatory effects and matrix metallopeptidase-1 (MMP-1)-inhibitory effects of CSE on PM-induced skin, NO and MMP-1 expressions were measured using an enzyme-linked immunosorbent assay (ELISA) kit. Also, the effects of CSE was investigated the induction of IL-8 and TNF-α treated PM on reconstructed human full thickness skin models.Results: It was observed that CSE decreased NO production in PM-induced RAW 264.7 cells without cytotoxicity. In addition, CSE decreased the expression of MMP-1 in PM-induced cells in a dose-dependent manner. CSE decreased IL-8 and TNF-α production in a PM-reconstructed human skin model.Conclusion: These results indicate that CSE could be used as a cosmetic material to induce anti-inflammation and inhibition of MMP-1 in PM-induced skin.


Marine Drugs ◽  
2020 ◽  
Vol 18 (7) ◽  
pp. 340
Author(s):  
Yingjuan Liu ◽  
Xiaolin Wu ◽  
Weihua Jin ◽  
Yunliang Guo

The water-soluble polysaccharide EP2, from Enteromorpha prolifera, belongs to the group of polysaccharides known as glucuronoxylorhamnan, which mainly contains glucuronic acid (GlcA), xylose (Xyl), and rhamnose (Rha). The aim of this study was to detect the immunomodulatory effects of EP2 on RAW 264.7 macrophages and cyclophosphamide (CYP)-induced immunosuppression mouse models. The cells were treated with EP2 for different time periods (0, 0.5, 1, 3, and 6 h). The results showed that EP2 promoted nitric oxide production and up-regulated the expression of pro-inflammatory cytokines, such as IL-1β, IL-6, and TNF-α, in a time-dependent manner. Furthermore, we found that EP2-activated iNOS, COX2, and NLRP3 inflammasomes, and the TLR4/MAPK/NF-κB signaling pathway played an important role. Moreover, EP2 significantly increased the body weight, spleen index, thymus index, inflammatory cell counts, and the levels of IL-1β, IL-6, and TNF-α in CYP-induced immunosuppression mouse models. These results indicate that EP2 might be a potential immunomodulatory drug and provide the scientific basis for the comprehensive utilization and evaluation of E. prolifera in future applications.


2010 ◽  
Vol 298 (4) ◽  
pp. C929-C941 ◽  
Author(s):  
Nilesh M. Dagia ◽  
Gautam Agarwal ◽  
Divya V. Kamath ◽  
Anshu Chetrapal-Kunwar ◽  
Ravindra D. Gupte ◽  
...  

A promising therapeutic approach to diminish pathological inflammation is to inhibit the increased production and/or biological activity of proinflammatory cytokines (e.g., TNF-α, IL-6). The production of proinflammatory cytokines is controlled at the gene level by the activity of transcription factors, such as NF-κB. Phosphatidylinositol 3-kinase (PI3K), a lipid kinase, is known to induce the activation of NF-κB. Given this, we hypothesized that inhibitors of PI3K activation would demonstrate anti-inflammatory potential. Accordingly, we studied the effects of a preferential p110α/γ PI3K inhibitor (compound 8C; PIK-75) in inflammation-based assays. Mechanism-based assays utilizing human cells revealed that PIK-75-mediated inhibition of PI3K activation is associated with dramatic suppression of downstream signaling events, including AKT phosphorylation, IKK activation, and NF-κB transcription. Cell-based assays revealed that PIK-75 potently and dose dependently inhibits in vitro and in vivo production of TNF-α and IL-6, diminishes the induced expression of human endothelial cell adhesion molecules (E-selectin, ICAM-1, and VCAM-1), and blocks human monocyte-endothelial cell adhesion. Most importantly, PIK-75, when administered orally in a therapeutic regimen, significantly suppresses the macroscopic and histological abnormalities associated with dextran sulfate sodium-induced murine colitis. The efficacy of PIK-75 in attenuating experimental inflammation is mediated, at least in part, due to the downregulation of pertinent inflammatory mediators in the colon. Collectively, these results provide first evidence that PIK-75 possesses anti-inflammatory potential. Given that PIK-75 is known to exhibit anti-cancer activity, the findings from this study thus reinforce the cross-therapeutic functionality of potential drugs.


2013 ◽  
Vol 2013 ◽  
pp. 1-6 ◽  
Author(s):  
Eun-Jin Yang ◽  
Young Min Ham ◽  
Kyong-Wol Yang ◽  
Nam Ho Lee ◽  
Chang-Gu Hyun

During our ongoing screening program designed to determine the anti-inflammatory potential of natural compounds, we isolated sargachromenol fromSargassum micracanthum. In the present study, we investigated the anti-inflammatory effects of sargachromenol on lipopolysaccharide (LPS)-induced inflammation in murine RAW 264.7 macrophage cells and the underlying mechanisms. Sargachromenol significantly inhibited the LPS-induced production of nitric oxide (NO) and prostaglandin E2(PGE2) in a dose-dependent manner. It also significantly inhibited the protein expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) in a dose-dependent manner in LPS-stimulated macrophage cells. Further analyses showed that sargachromenol decreased the cytoplasmic loss of inhibitorκBα(IκBα) protein. These results suggest that sargachromenol may exert its anti-inflammatory effects on LPS-stimulated macrophage cells by inhibiting the activation of the NF-κB signaling pathway. In conclusion, to our knowledge, this is the first study to show that sargachromenol isolated fromS. micracanthumhas an effective anti-inflammatory activity. Therefore, sargachromenol might be useful for cosmetic, food, or medical applications requiring anti-inflammatory properties.


2020 ◽  
Vol 37 (2) ◽  
pp. 88-93
Author(s):  
Na Young Jo

Background: The purpose of this study was to investigate whether Sibseonsan (SSS) is an effective anti-inflammatory, anti-wrinkling, and whitening agent.Methods: To determine whether SSS had an anti-inflammatory effect, a murine macrophage cell line was used (RAW 264.7) and production of DPPH, NO, TNF-α, and PGE2 were measured. To ascertain potential anti-wrinkle effects of SSS in these cells, collagenase and elastase production were measured. To verify whether SSS had a whitening effect, tyrosinase activity and DOPA staining were performed using a melanoma cell line (B16/F10).Results: There was no significant reduction in survival of SSS-treated RAW 264.7 cells, up to 400 μg/mL. Free radical scavenging (23.96 ± 1.85%) was observed in RAW 264.7 cells treated with SSS at a concentration of 400 μg/mL. The SSS treatment group (400 μg/mL) significantly inhibited NO production compared with the LPS stimulated treatment group. The SSS treatment of macrophage cells appeared to reduce production of TNF-α in a concentration dependent manner. There was a significant reduction in the concentration of PGE<sub>2</sub> by about 25% in the SSS treatment (400 μg/mL) group (<i>p</i> = 0.05). Compared with the control, the production of collagenase and elastase in B16/F10 cells treated with SSS (400 μg/mL) was greater by 26.37% and 45.71%, respectively. The SSS treatment (400 μg/mL) group showed a significant reduction by about 17% in tyrosinase production in B16/F10 cells. The SSS treatment group showed little change in DOPA staining.<br>Conclusion: SSS extract may be useful for the treatment and prevention of inflammatory diseases and may have anti-wrinkle and whitening effects. These results may support the use of SSS in clinical practice.


2021 ◽  
Vol 2021 ◽  
pp. 1-10
Author(s):  
Zhenyu Fan ◽  
Liangliang Cai ◽  
Yage Wang ◽  
Qiuyan Zhu ◽  
Shengnan Wang ◽  
...  

Isatidis Radix, the dried root of Isatidis indigotica Fort, is a traditional heat-clearing and detoxicating herb, which has the antiviral and anti-inflammatory activity and immune regulation. It has been widely used to treat cold, fever, sore throat, mumps, and tonsillitis in clinics. A previous study demonstrated that the acidic fraction of Isatidis Radix (RIAF) had strong anti-inflammatory activity, but the mechanism of action was not well elucidated. Lipopolysaccharide- (LPS-) induced RAW264.7 cells were employed to observe the anti-inflammatory activity of RIAF. The level of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), nitric oxide (NO), prostaglandin E2 (PGE2), and interleukin-6 (IL-6) was determined by enzyme-linked immunosorbent assay kits. Western blot was performed to quantify the expression of extracellular signal-regulated kinase (ERK) 1/2, c-jun NH2-termianl kinase (JNK), p38, inducible NO synthetase (iNOS), cyclooxygenase (COX)-2, andnuclear factor-κB (NF-κB). Immunofluorescence assay and electrophoretic mobility shift assay (EMSA) were used to quantify the translocation and the binding-DNA activity of NF-κB. RIAF could inhibit the secretion of inflammatory cytokines (PGE2, IL-6, IL-1β, and NO, other than TNF-α) in a dose-dependent manner. Further investigation showed that the expression of iNOS and COX-2 induced by LPS were downregulated by treatment with RIAF. Meanwhile, data from the signal pathway exhibited that RIAF significantly suppressed the phosphorylation of ERK1/2, JNK, and p38 and reduced the translocation of NF-κB from the cytoplasm to nucleus, as well as the binding-DNA activity. The anti-inflammatory mechanism of action of RIAF was to reduce inflammation-associated gene expression (iNOS, COX-2, IL-1β, IL-6) by regulating the phosphorylation of the mitogen-activated protein kinases (MAPK) pathway and interventing the activation of the NF-κB pathway, which partly illustrated the basis of treatment of Isatidis Radix on cold, fever, sore throat, mumps, and tonsillitis in clinics.


Author(s):  
Sangeeta Mohanty ◽  
Abhisek Pal ◽  
V Badireenath Konkimalla ◽  
Sudam Chandra Si

Objective: The objective of this study was to establish the anti-inflammatory activity of sulforaphane (SFN) in different acute and subchronic models of inflammation. Methods: The anti-inflammatory activity of SFN was evaluated by the secretion of proinflammatory cytokines in rat peripheral blood mononuclear cells (interleukin-6 [IL-6] and tumor necrosis factor alpha [TNF-α]) which are important mediators of inflammation as determined by enzyme-linked immunosorbent assay. Furthermore, paw volume was determined in various acute models of inflammation, and percentage inhibition of granuloma tissue was assessed by cotton pellet-induced granuloma model. From serum, serum glutamic pyruvic transaminase, serum glutamic oxaloacetic transaminase, and alkaline phosphatase levels were determined which is followed by assay for estimation of antioxidants such as superoxide dismutase (SOD), catalase, and glutathione (GSH). Results: SFN showed significant anti-inflammatory activity against paw edema induced by carrageenin/histamine/egg-albumin. A remarkable control in inflammation was observed most notably at the highest test dose of 5 mg/kg in the subchronic granuloma model. In addition, the release of inflammatory cytokines such as IL-6 and TNF-α which is responsible for inflammatory activity gets attenuated by SFN (∗p<0.05; ∗∗p< 0.01). Moreover, toxic control rats showed significant decreased levels of GSH, catalase, and SOD and increased the level of serum hepatic enzymes which gets reversed by SFN in dose-dependent manner. Conclusions: The present findings demonstrated that SFN can recover inflammation by inhibiting TNF-α and IL-6 in inflammation process.


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