Enzymatic hydrolysis on protein and β-glucan content of Sang-yod rice bran hydrolysates and their anti-inflammatory activity on RAW 264.7 cells

Background: Research focusing on the improvement of the utilization of rice bran is increasing due to its nutritional properties. Several biological activities of rice bran hydrolysates and its constituents have been reported. Sang-yod rice, a local rice variety in Southern of Thailand, is a pigmented rice. Furthermore, its bran has high nutritive value and health beneficial components. Accordingly, there is growing interest in transforming this by-product into a functional food ingredient.Objective: To investigate the effect of enzymatic hydrolysis processes on the digestion of protein and β-glucan and evaluate anti-proinflammatory properties of selected hydrolysates on RAW 264.7 macrophage cells.Method: Sang-yod rice bran hydrolysates were obtained using a single or co-enzymatic hydrolysis process and sequential hydrolysis process using amyloglucosidase and protease G6. Effects of enzyme concentration (3-5% v/w) and hydrolysis duration (30, 60, and 120 min) on soluble protein and β-glucan contents of obtained rice bran hydrolysates were evaluated. The selected rice bran hydrolysates were evaluated for their cell viability and inhibition against NO and pro-inflammatory cytokines generation on RAW 264.7 mouse macrophage cell lines. Results: Protein content (0.59-3.37 %) of the rice bran hydrolysates (RBHs) was increased by increasing of enzyme concentration (3-5% v/w) and hydrolysis time (60-120 min). However, the β-glucan content (0.88-4.63%) of RBHs decreased with the increase of those parameters. The RBHs derived by the sequential process using 5% v/w enzyme concentration and 60 min hydrolysis time gave high protein (3.23%) and high β-glucan (4.02%) contents. The hydrolysates with high amount of protein and/or β-glucan contents demonstrated no cytotoxicity against RAW 264.7 cells at concentration range of 100-2,000 μg/ml. Additionally, they demonstrated NO inhibition and pro-inflammatory inhibition ranges of 49.09-71.63% and 9.37-71.96% respectively. Generation of TNF-α, IL-6, and IL-1β cytokines was inhibited differently by the selected RBHs. Conclusion: Pre-digestion of Sang-yod rice bran with amyloglucosidase followed with co-hydrolysis of amyloglucosidase and protease G6 of the sequential hydrolysis process was the most effective process to release β-glucan and protein from of rice bran. The hydrolysate obtained from the process using enzyme concentration at 5%v/w and 60 min hydrolysis duration of each stage had the highest soluble β-glucan and protein content. Moreover, the process provided the hydrolysates with potential anti-inflammatory properties on nitric oxide inhibition and pro-inflammatory cytokines inhibition on RAW 264.7 macrophage cell line. Keywords: Sang-yod rice, Rice bran hydrolysate, β-glucan, Enzymatic hydrolysis, Anti-inflammatory activity

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Agrimonia pilosa Ledeb Root Extract: Anti-Inflammatory Activities of the Medicinal Herb in LPS-Induced Inflammation

The American Journal of Chinese Medicine ◽

10.1142/s0192415x20500949 ◽

2020 ◽

Vol 48 (08) ◽

pp. 1875-1893

Author(s):

Da-Sol Kim ◽

Kyoung-Eun Park ◽

Yeon-Ju Kwak ◽

Moon-Kyoung Bae ◽

Soo-Kyung Bae ◽

...

Keyword(s):

Porphyromonas Gingivalis ◽

Inflammatory Cytokines ◽

Mapk Signaling ◽

The Body ◽

Raw 264.7 Cells ◽

Root Extract ◽

Raw 264.7 ◽

Cox 2 ◽

Anti Inflammatory ◽

Pro Inflammatory Cytokines

Inflammation regulation is essential for maintaining healthy functions and normal homeostasis of the body. Porphyromonas gingivalis (P. gingivalis) is a gram-negative anaerobic bacterium and a major pathogen that causes oral inflammation and other systemic inflammations. This study aims to examine the anti-inflammatory effects of Agrimonia pilosa Ledeb root extracts (APL-ME) in Porphyromonas gingivalis LPS-induced RAW 264.7 cells and find anti-inflammatory effect compounds of APL-ME. The anti-inflammatory effects of APL-ME were evaluated anti-oxidant activity, cell viability, nitrite concentration, pro-inflammatory cytokines (interleukin-1[Formula: see text], interleukin-6, tumor necrosis factor (TNF)-[Formula: see text], and anti-inflammatory cytokine (interleukin-10 (IL-10)). Also, Inflammation related genes and proteins, cyclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS), expression were decreased by APL-ME and mitogen-activated protein kinase (MAPK) signaling proteins expression was regulated by APL-ME. Liquid chromatography-mass spectrometer (LC/MS)-MS analysis results indicated that several components were detected in APL-ME. Our study indicated that APL-ME suppressed nitrite concentrations, pro-inflammatory cytokines such as IL-1[Formula: see text], IL-6 and TNF-[Formula: see text] in P. gingivalis LPS induced RAW 264.7 cells. However, IL-10 expression was increased by ALP-ME. In addition, protein expressions of COX-2 and iNOS were inhibited APL-ME extracts dose-dependently. According to these results, APL-ME has anti-inflammatory effects in P. gingivalis LPS induced RAW 264.7 cells.

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Anti-Inflammatory Effects of 6,7-Dihydroxy-4-Methylcoumarin on LPS-Stimulated Macrophage Phosphorylation in MAPK Signaling Pathways

Natural Product Communications ◽

10.1177/1934578x211020970 ◽

2021 ◽

Vol 16 (5) ◽

pp. 1934578X2110209

Author(s):

Yun Sil Kang ◽

You Chul Chung ◽

Jung No Lee ◽

Bong Seok Kim ◽

Chang-Gu Hyun

Keyword(s):

Signaling Pathways ◽

Inflammatory Cytokines ◽

Inflammatory Diseases ◽

Mapk Signaling ◽

Raw 264.7 Cells ◽

Inflammatory Factors ◽

Raw 264.7 ◽

Dependent Manner ◽

Anti Inflammatory ◽

Pro Inflammatory Cytokines

Coumarin derivatives, such as esculetin, have various physiological functions, including antioxidant, anti-inflammatory, antibacterial, antiviral, and anti-cancer. 6,7-Dihydroxy-4-methylcoumarin (6,7-DH-4MC) is a derivative of esculetin, and its anti-inflammatory effect and mechanism in macrophages have not been studied. In this study, the anti-inflammatory activity of 6,7-DH-4MC was evaluated by measuring the expression of inflammatory factors (NO and PGE2) and pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α) in LPS-stimulated RAW 264.7 macrophages. The results revealed that 6,7-DH-4MC significantly reduced NO levels and PGE2 expression without inducing cytotoxicity; it was confirmed that the inhibition of NO and PGE2 expression was related to iNOS and COX-2 downregulation in response to 6,7-DH-4MC treatment. Moreover, 6,7-DH-4MC decreased the levels of pro-inflammatory cytokines, such as IL-1β and IL-6, in a dose-dependent manner. Mechanistic studies revealed reduced phosphorylation of ERK and p38-MAPK upon 6,7-DH-4MC treatment. Furthermore, the degradation of IκB-α and phosphorylation of NF-κB in cells treated with LPS were interrupted by 6,7-DH-4MC treatment. These results suggest that 6,7-DH-4MC is a potential therapeutic agent for inflammatory diseases. To the best of our knowledge, this is the first report demonstrating the anti-inflammatory effects of 6,7-DH-4MC in RAW 264.7 cells via MAPK and NF-κB signaling pathways.

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Anti-Inflammatory Effects of 6-Methylcoumarin in LPS-Stimulated RAW 264.7 Macrophages via Regulation of MAPK and NF-κB Signaling Pathways

Molecules ◽

10.3390/molecules26175351 ◽

2021 ◽

Vol 26 (17) ◽

pp. 5351

Author(s):

Jin-Kyu Kang ◽

You-Chul Chung ◽

Chang-Gu Hyun

Keyword(s):

Nitric Oxide ◽

Inflammatory Cytokines ◽

Mitogen Activated Protein Kinase ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Anti Inflammatory ◽

Pro Inflammatory Cytokines

Persistent inflammatory reactions promote mucosal damage and cause dysfunction, such as pain, swelling, seizures, and fever. Therefore, in this study, in order to explore the anti-inflammatory effect of 6-methylcoumarin (6-MC) and suggest its availability, macrophages were stimulated with lipopolysaccharide (LPS) to conduct an in vitro experiment. The effects of 6-MC on the production and levels of pro-inflammatory cytokines (interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α) and inflammatory mediators (nitric oxide (NO), prostaglandin E2 (PGE2)) in LPS-stimulated RAW 264.7 cells were examined. The results showed that 6-MC reduced the levels of NO and PGE2 without being cytotoxic. In addition, it was demonstrated that the increase in the expression of pro-inflammatory cytokines caused by LPS stimulation, was decreased in a concentration-dependent manner with 6-MC treatment. Moreover, Western blot results showed that the protein levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), which increased with LPS treatment, were decreased by 6-MC treatment. Mechanistic studies revealed that 6-MC reduced the phosphorylation of the mitogen-activated protein kinase (MAPK) family and IκBα in the MAPK and nuclear factor-kappa B (NF-κB) pathways, respectively. These results suggest that 6-MC is a potential therapeutic agent for inflammatory diseases that inhibits inflammation via the MAPK and NF-κB pathways.

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Assessment of Anti-Inflammatory and Antioxidant Effects of Citrus unshiu Peel (CUP) Flavonoids on LPS-Stimulated RAW 264.7 Cells

Plants ◽

10.3390/plants10102209 ◽

2021 ◽

Vol 10 (10) ◽

pp. 2209

Author(s):

Adhimoolam Karthikeyan ◽

Hun Hwan Kim ◽

Vetrivel Preethi ◽

Mohammad Moniruzzaman ◽

Ki Ho Lee ◽

...

Keyword(s):

Biologically Active ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Citrus Unshiu ◽

Raw 264.7 Macrophages ◽

Tnf Α ◽

Citrus Unshiu Peel ◽

Anti Inflammatory ◽

Antioxidant Effects ◽

Pro Inflammatory Cytokines

Citrus unshiu is a popular medicinal herb in several Asian countries, in particular South Korea. C. unshiu peel (CUP) has several biologically active compounds, including flavonoids. Hence, this research aimed to label the flavonoids from CUP by HPLC-MS/MS analysis and examine their anti-inflammatory and antioxidant potential on LPS-stimulated RAW 264.7 macrophages. A total of four flavonoids (Rutin, naringin, hesperidin, and poncirin) were characterized, and their contents were quantified from CUP. It showed that the naringin is rich in CUP. Further, treatment with the flavonoids at concentrations of 2.5 and 5 μg/mL had no effect on the cell viability of RAW 264.7 macrophages. On the other hand, it decreased the production and expression of inflammatory mediators and pro-inflammatory cytokines such as NO, PGE2, TNF-α, IL-1β, iNOS, and COX2 in the LPS-stimulated RAW 264.7 macrophages. In addition, flavonoids treatment inhibited the NF-κB activation by downregulating the p-p65 and p-IκBα proteins expression. Furthermore, reactive oxygen species (ROS) production considerably decreased at the same concentrations while antioxidant enzyme activity increased in the LPS-stimulated RAW 264.7 macrophages. Collectively, our results show that CUP flavonoids have the potential to decrease inflammation and oxidative damage.

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Decursinol Angelate Inhibits LPS-Induced Macrophage Polarization through Modulation of the NFκB and MAPK Signaling Pathways

Molecules ◽

10.3390/molecules23081880 ◽

2018 ◽

Vol 23 (8) ◽

pp. 1880 ◽

Cited By ~ 15

Author(s):

Salman Islam ◽

Jung Lee ◽

Adeeb Shehzad ◽

Eun-Mi Ahn ◽

You Lee ◽

...

Keyword(s):

Map Kinase ◽

Signaling Pathways ◽

Inflammatory Cytokines ◽

Macrophage Polarization ◽

Raw 264.7 Cells ◽

M2 Macrophage ◽

Raw 264.7 ◽

Decursinol Angelate ◽

Anti Inflammatory ◽

Pro Inflammatory Cytokines

Inflammation is considered the root cause of various inflammatory diseases, including cancers. Decursinol angelate (DA), a pyranocoumarin compound obtained from the roots of Angelica gigas, has been reported to exhibit potent anti-inflammatory effects. In this study, the anti-inflammatory effects of DA on the MAP kinase and NFκB signaling pathways and the expression of pro-inflammatory cytokines were investigated in phorbol 12-myristate 13-acetate (PMA)-activated human promyelocytic leukemia (HL-60) and lipopolysaccharide (LPS)-stimulated macrophage (Raw 264.7) cell lines. PMA induced the activation of the MAP kinase-NFκB pathway and the production of pro-inflammatory cytokines in differentiated monocytes. Treatment with DA inhibited the activation of MAP kinases and the translocation of NFκB, and decreased the expression and exogenous secretion of IL-1β and IL-6. Furthermore, LPS-stimulated Raw 264.7 cells were found to have increased expression of M1 macrophage-associated markers, such as NADPH oxidase (NOX) and inducible nitric oxide synthase (iNOS), and the M2 macrophage-associated marker CD11b. LPS also activated pro-inflammatory cytokines and Erk-NFκB. Treatment with DA suppressed LPS-induced macrophage polarization and the inflammatory response by blocking Raf-ERK and the translocation of NFκB in Raw 264.7 cells. Treatment with DA also inhibited the expression of pro-inflammatory cytokines, such as IL-1β and IL-6, NOX, and iNOS in Raw 264.7 cells. These results suggest that DA has the potential to inhibit macrophage polarization and inflammation by blocking the activation of pro-inflammatory signals. These anti-inflammatory effects of DA may contribute to its potential use as a therapeutic strategy against various inflammation-induced cancers.

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5-Hydroxymethylfurfural Mitigates Lipopolysaccharide-Stimulated Inflammation via Suppression of MAPK, NF-κB and mTOR Activation in RAW 264.7 Cells

Molecules ◽

10.3390/molecules24020275 ◽

2019 ◽

Vol 24 (2) ◽

pp. 275 ◽

Cited By ~ 13

Author(s):

Fanhui Kong ◽

Bae Lee ◽

Kun Wei

Keyword(s):

Nitric Oxide ◽

Inflammatory Cytokines ◽

Mammalian Target Of Rapamycin ◽

Inflammatory Activity ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Anti Inflammatory ◽

Pro Inflammatory Cytokines

5-Hydroxymethylfurfural (5-HMF) is found in many food products including honey, dried fruits, coffee and black garlic extracts. Here, we investigated the anti-inflammatory activity of 5-HMF and its underlying mechanisms in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. 5-HMF pretreatment ranging from 31.5 to 126.0 μg/mL reduced the production of nitric oxide (NO), prostaglandin E2 (PGE2) and pro-inflammatory cytokines (TNF-α, IL-1β and IL-6) in a concentration-dependent manner in LPS-stimulated cells. Moreover, 5-HMF-pretreated cells significantly down-regulated the mRNA expression of two major inflammatory mediators, nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) and suppressed the production of pro-inflammatory cytokines, as compared with the only LPS-stimulated cells. 5-HMF suppressed the phosphorylation of extracellular regulated protein kinases (ERK1/2), c-Jun N-terminal kinase (JNK), IκBα, NF-κB p65, the mammalian target of rapamycin (mTOR) and protein kinase B (Akt). Besides, 5-HMF was proved to inhibit NF-κB p65 translocation into nucleus to activate inflammatory gene transcription. These results suggest that 5-HMF could exert the anti-inflammatory activity in the LPS-induced inflammatory response by inhibiting the MAPK, NF-κB and Akt/mTOR pathways. Thus, 5-HMF could be considered as a therapeutic ingredient in functional foods.

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Effect of Fractions, and Compounds from Typha capensis in LPS-Stimulated Raw 264.7 Cells. Pro and Anti-inflammatory Cytokines

Current Journal of Applied Science and Technology ◽

10.9734/cjast/2021/v40i1431399 ◽

2021 ◽

pp. 20-27

Author(s):

Moise Ondua

Keyword(s):

Inflammatory Cytokines ◽

Interleukin 1 ◽

Molecular Targets ◽

Traditional Healers ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Necrosis Factor Alpha ◽

Tnf Α ◽

Anti Inflammatory ◽

Pro Inflammatory Cytokines

Typha capensis is widely used by traditional healers to treat male fertility, venereal problems and inflammation. There are many molecular targets implicated in the inflammatory process: pro- and anti-inflammatory cytokines such as interleukin 1-β, IL-6, IL-10, IL-12p70, tumor necrosis factor alpha (TNF-α), and IL-8, and other proteins such as COX-2, and iNOS. In order to clarify the anti-inflammatory mechanism of action of compounds isolated from T. capensis, RAW 264.7 macrophages were activated by lipopolysaccharide and pre-treated with T. capensis isolated compounds. Lipopolysaccharide-stimulated RAW macrophages after treatment with T. capensis crude acetone extract resulted in decreasing expression of pro-inflammatory cytokines (TNF-α, IL-6,) and increased expression of immunomodulatory cytokine IL-12 P 70. Isorhamnetin-3-O-β-D-glucoside and isorhamnetin 3-O rutinoside increased the expression of pro-inflammatory cytokines TNF-α, but failed to reduce the expression of IL-1β and TNF-α. Isorhamnetin-3-O-β-D-glucoside and isorhamnetin 3-O rutinoside increased the expression of immunomodulatory cytokine IL-12p70. Isorhamnetin-3-O-β-D-glucoside increased the expression of the anti-inflammatory cytokine IL-10 compared to quercetin and LPS-stimulated macrophages. The effect of isorhamnetin 3-O-rutinoside and isorhamnetin-3-O-β-D-glucoside on molecular targets of inflammation may provide support for the use of T. capensis by traditional healers against inflammation.

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Effect of Ultrasound-Assisted on Sequential Peracetic Acid and Alkaline Peroxide Pretreatment to The Enzymatic Hydrolysis of Oil Palm Empty Fruit Bunch

10.21203/rs.3.rs-284744/v1 ◽

2021 ◽

Author(s):

Dwini Normayulisa Putri ◽

Meka Saima Perdani ◽

Masafumi Yohda ◽

Tania Surya Utami ◽

Muhamad Sahlan ◽

...

Keyword(s):

Enzymatic Hydrolysis ◽

Oil Palm ◽

Peracetic Acid ◽

Enzyme Concentration ◽

Reducing Sugar ◽

Empty Fruit Bunch ◽

Hydrolysis Time ◽

Alkaline Peroxide ◽

Hydrolysis Process ◽

Hydrolysis Of

Abstract Enzymatic hydrolysis of oil palm empty fruit bunch (OPEFB) that has been pretreated by modified pretreatment has been investigated in this study. The OPEFB used was pretreated by using sequential peracetic acid – alkaline peroxide solution. As the modification method, the assistance of pretreatment by ultrasound was conducted, in order to increase the enzyme accessibility. Therefore, it enhances the production of reducing sugar on the hydrolysis process. Prior to hydrolysis process, OPEFB was initially treated by using peracetic acid solution, comprise of CH3COOH (> 99%) and H2O2 (30% w/w), assisted by ultrasound for 3 hours at 35oC. Afterwards, OPEFB was treated by using alkaline peroxide solution, comprise of NaOH (40% w/w) and H2O2 (35% w/w), assisted by ultrasound for 10 hours at 35oC. OPEFB that has been pretreated was then subjected to enzymatic hydrolysis process using cellulase enzyme, in order to convert cellulose content into reducing sugar. Enzymatic hydrolysis was carried out at 50oC in a shaker incubator with 150 rpm for 48 hours. In this study, the effect of different enzyme concentration and hydrolysis time towards the sugar concentration in modified-pretreated OPEFB was observed and analyzed. Three different concentrations of enzyme were used, including 1.25, 2.5, and 5 g/L, and reducing sugar concentrations were analyzed at 30 and 45 minutes, and 1, 2, 4, 6, 24, 30, and 48 hours. Based on results, enzyme concentration has a significant effect to the production of reducing sugar. The reducing sugar concentrations obtained at the end of the hydrolysis process were 8.48, 11.06, 19.16 g/L, at the enzyme concentrations of 1.25, 2.5, and 5 g/L, respectively. At any hydrolysis time, the highest sugar concentration has been achieved on the highest enzyme concentration of 5 g/L. Moreover, the effective hydrolysis time were achieved at 6 hours, at all concentration of enzyme, since the production of reducing sugar were insignificant after 6 hours. This study showed an increase in reducing sugar production by 8.25% in the hydrolysis process using OPEFB pretreated by modified pretreatment compared to the non-modified pretreatment.

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