• Journal of the Korean Applied Science and Technology
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• v.38 no.1
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• pp.309-317
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• 2021

This study was designed to examine the cell cytotoxic, anti-inflammatory, and antioxidant activity for raw material of Beta vulagaris tea. Antioxidative ability was evaluated by bioassays using DPPH and ABTS radical scavenging activity. Cell viability was assessed by MTT assay using RAW 264.7 cells, and investigated production levels of reactive oxide speies, and inflammatory meditors(i.e., nitric oxide, tumor necrosis factor-α, and interleukin-6) in LPS-induced RAW 264.7 cells. As a results, DPPH and ABTS raidcal scavenging activity were increased in a dose-dependent manner, and confirmed no cytotoxicity in all concentration. Also, it was significantly decreased level of ROS, NO, IL-6, and TNF-a in LPS-induced RAW 264.7 cells. Therefore these results suggest Beta vulagaris has considerable potential as a raw material of leached tea with safe anti-oxidative and anti-inflammatory effects.

• Journal of Life Science
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• v.32 no.2
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• pp.125-134
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• 2022

This study investigated whether or not the major bioactive compounds of Noni (Morinda citrifolia) are involved in anti-inflammatory activity through the JAK/STAT upper signaling pathway in RAW 264.7 cells. The experimental results show that the M. citrifolia ethyl acetate fraction (Mc-EtOAc) obtained by sequential fractionation with organic solvents from the plant's dried fruits exhibits the highest antioxidant activity. In addition, the cytoprotective effects of Mc-EtOAc against H₂O₂-induced oxidative stress in the RAW 264.7 cells suppressed cytotoxicity in a dose-dependent manner. The group pretreated with Mc-EtOAc at a concentration of 240 ㎍/ml showed higher cell viability of 84.5%, compared to 71.6% in the LPS-treated group, and LPS-induced NO production decreased to half the amount in the positive control group. Mc-EtOAc treatment also led to a significant dose-dependent reduction in iNOS expression. Although COX-2 expression was increased by 300% following LPS induction, it was significantly decreased in a dose-dependent manner by pretreatment with Mc-EtOAc at concentrations of 120 and 240 ㎍/ml. An inhibition of the mRNA expression of pro-inflammatory cytokines IL-1β and TNF-α was observed. The investigation also revealed that the phosphorylation levels of pJAK1 and pSTAT3 in LPS-induced RAW 264.7 cells were significantly reduced by Mc-EtOAc treatment.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.20 no.2 s.33
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• pp.68-76
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• 2007

Objectives : This experimental study was performed to investigate the effects of Sulfur extract on anti-inflammation and anti-Propionibacterium acnes. Methods : The cytotoxicity of Sulfur extract about viability of Raw 264.7 cell were tested using a colorimetric tetrazolium assay(MTT assay). To investigate the anti-inflammation effects of Sulfur extract on LPS-induced macrophage Raw 264.7 cell, we used ELISA kit and Western blots. We evaluated anti-oxidation effects of Sulfur extract on HaCaT cell by Enzyme recycling method. And we investigated the inhibitory effects of Sulfur extract on Propionibactrium acnes using paper disk diffusion method. Results: 1. Sulfur extract has a little cytotoxicity in Raw 264.7 cell. 2. Concentration of $100\;{\mu}g/m{\ell}$ Sulfur extract inhibited the production of NO in the Raw 264.7 cell stimulated with LPS. 3. Sulfur extract showed a oxidation inhibition effect by decreasing the DPPH radicals. 4. Sulfur extract has not the significant inhibition effect of Propionibactrium acnes. Conclusions: These results indicate that Sulfur extract has anti-inflammation and anti-oxidation effects. If further study is performed, the use of Sulfur extract will be valuable and benificial in the therapy of Propionibactrium acnes.

• Archives of Pharmacal Research
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• v.29 no.3
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• pp.218-223
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• 2006

Erythropoietin (EPO), a hematopoietic factor, is required for normal erythrocyte developments, but it has been demonstrated to have many other functions, and its receptor is localized in other tissues. In the present study, we investigated whether EPO can promote other cell proliferation and possible molecular mechanisms. EPO restored the inhibition of the RAW264.7 and PC12 cell growth by fetal bovine serum (FBS) withdrawal in a dose dependent manner, but not that of other cell types tested. The restoring effect of EPO was completed when the RAW264.7 cells were cultured in the medium containing as low as 3% of FBS, and 10 U/mL EPO could replace FBS. The restoring effect of EPO in the RAW264.7 cells was associated with the increased of c-Fos and c-Jun expression as well as AP-1 activation. These data demonstrate that EPO can stimulate RAW264. 7 cell as well as PC12 cell growth even when the cells were cultured without FBS or in the presence of small amounts of FBS in the medium, and this stimulating effect is associated with the activation of AP-1 transcription factor.

• Archives of Pharmacal Research
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• v.29 no.10
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• pp.890-897
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• 2006

In inflammatory responses, induction of cytokines and other immune regulator genes in macrophages by pathogen-associated signal such as lipopolysaccharide (LPS) plays a crucial role. In this study, the gene expression profile changes by LPS treatment in the macrophage/monocyte lineage cell line RAW264.7 was investigated. A 60-mer oligonucleotide microarray of which probes target 32381 mouse genes was used. A reverse transcription-in vitro translation labeling protocol and a chemileuminescence detection system were employed. The mRNA expression levels in RAW264.7 cells treated for 6 h with LPS and the control vehicle were compared. 747 genes were up-regulated and 523 genes were down-regulated by more than 2 folds. 320 genes showing more than 4-fold change by LPS treatment were further classified for the biological process, molecular function, and signaling pathway. The biological process categories that showed high number of increased genes include the immunity and defense, the nucleic acid metabolism, the protein metabolism and modification, and the signal transduction process. The chemokine-cytokine signaling, interleukin signaling, Toll receptor signaling, and apoptosis signaling pathways involved high number of genes differentially expressed in response to LPS. These expression profile data provide more comprehensive information on LPS-target genes in RAW264.7 cells, which will be useful in comparing gene expression changes induced by extracts and compounds from anti-inflammatory medicinal herbs.

• Preventive Nutrition and Food Science
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• v.16 no.4
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• pp.299-306
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• 2011

The present study was designed to evaluate the inhibitory effects of fermented Liriope platyphylla extract on the production of inflammation-related mediators (NO, ROS, NF-${\kappa}B$, iNOS and COX-2) and pro-inflammatory cytokines (TNF-${\alpha}$, IL-$1{\beta}$, IL-6) in lipopolysaccharide-stimulated RAW 264.7 macrophages. Freeze-dried Liriope platyphylla was fermented by Saccharomyces cerevisiae and extracted with 70% ethanol. In lipopolysaccharide-stimulated macrophage cells, the treatment with fermented Liriope platyphylla extract decreased the generation of intracellular reactive oxygen species dose-dependently and increased antioxidant enzyme activities, including superoxide dismutase, catalase and glutathione peroxidase. Fermented Liriope platyphylla extract also inhibited NO production in lipopolysaccharide-stimulated RAW 264.7 cell. The expressions of NF-${\kappa}B$, iNOS, COX-2 and pro-inflammatory cytokines were inhibited by the treatment with fermented Liriope platyphylla extract. Thus, this study shows the fermented Liriope platyphylla extract could be effective at inhibiting the inflammation process.

• Microbiology and Biotechnology Letters
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• v.44 no.3
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• pp.383-390
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• 2016

Mesenchymal stem cells (MSCs) are multipotent stem cells that can be differentiated into a variety of cell types, including adipocytes, osteoblasts, chondrocytes, β-pancreatic islet cells, and neuronal cells. MSCs have been reported to exhibit immunomodulatory effects in many diseases. Many studies have reported that MSCs have distinct roles in modulating inflammatory and immune responses by releasing bioactive molecules. Exosomes are cell-derived vesicles present in biological fluids, including the blood, urine, and cultured medium of cell cultures. In this study, we investigated the immunomodulatory effects of mouse adipose tissue-derived MSCs (mAD-MSCs), cultured medium (MSC-CM) of mAD-MSCs, and mAD-MSC-derived exosomes (MSC-Exo) on lipopolysaccharide (LPS)-induced RAW 264.7 cells. We observed that the expression levels of IL-1β, TNF-α, and IL-10 were significantly increased in LPS-stimulated RAW 264.7 cells compared to those in LPS-unstimulated RAW 264.7 cells. Additionally, these values were significantly (p < 0.05) decreased in mAD-MSCs-RAW 264.7 cell co-culture groups, MSC-CM-treated groups, and MSC-Exo-treated groups. MSCs can modulate the immune system in part by secreting cytokines and growth factors. We observed that immunomodulatory factors such as IL-1β, TNF-α, and IL-10 were secreted by mAD-MSCs under co-culturing conditions of mAD-MSCs with activated RAW 264.7 cells. In addition, mAD-MSC-derived exosomes exhibited similar immunomodulatory effects in activated RAW 264.7 cells. Therefore, our results suggest that mAD-MSCs have an immunomodulatory function through indirect contact.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.12
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• pp.1671-1676
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• 2012

In this study, the immuno-enhancing activity of seed extracts were studied on the macrophage cell lines. We examined the effect of nine seed extracts on nitric oxide (NO) production in RAW 264.7 cells and selected four highly-effective seed candidates (Fagopyrum esculentum, Taraxacum platycarpum, Impatiens balsamina, Helianthus annuus) for further immune-related studies. The effects of the four seed extracts on the production of immune-related cytokines in the RAW 264.7 macrophage cell line and proliferation of Molt-4 as a T cell line were investigated. The secretion of NO from the RAW 264.7 cells was increased up to 39 ${\mu}M$ by adding the seed extracts (25 ${\mu}g/mL$) compared to the control. Also, the secretion of tumor necrosis factor-alpha (TNF-${\alpha}$) was also increased up to 32 times by adding the seed extracts (25 ${\mu}g/mL$). Secretion of cytokines such as interleukin-1 beta (IL-$1{\beta}$), interleukin-6 (IL-6), and interleukin-10 (IL-10) was also increased and induced the proliferation of T cells compared to the control. In conclusion, these results suggest that four seed extracts provide beneficial immuno-enhancing effects for human health.

• Korean Journal of Acupuncture
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• v.26 no.3
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• pp.69-76
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• 2009

Objectives : 본 연구의 목적은 제습, 해독, 통리관절등의 효능이 있는 토복령이 RAW264.7 대식세포주에서 lipopolysaccharide(LPS)로 처치하여 생성되는 nitric oxide(NO)와 prostaglandin $E_2$($PGE_2$)에 억제작용이 있는지 관찰하고자 하는 것이다. Methods : 토복령이 RAW264.7 대식세포주에 세포독성이 유무를 관찰하기 위하여 농도별 MTT assay를 수행하여 세포생존율을 측정하였다. 또한 LPS로 염증유발된 RAW264.7 대식세포주에서 토복령의 농도별 처치가 NO 및 $PGE_2$ 생성억제에 미치는 영향을 관찰하고자 NO 및 $PGE_2$ assay를 수행하였다. Results : 토복령의 농도별 처치가 RAW264.7 대식세포주에서 세포독성을 유발하는지 MTT assay로 측정한 결과 세포독성은 관찰되지 않았으며, 토복령은 LPS처치로 인하여 증가된 NO 및 $PGE_2$ 생성을 통계학적으로 유의하게 감소시킴을 관찰하였다. Conclusions : 본 연구를 통하여 토복령은 RAW264.7 대식세포주에서 LPS로 유도된NO 및 $PGE_2$ 생성을 효과적으로 억제시킴으로써 추후 염증질환의 치료제로서 가능성을 확인하였다.

• The Journal of Korean Obstetrics and Gynecology
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• v.34 no.3
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• pp.15-28
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• 2021

Objectives: This study was designed to examine the anti-cancer activity by innate immunomodulating and anti-inflammatory effects of liriopis tuber water extract (LPE). Methods: Cell cytotoxicity was tested with 4T1 mouse mammary carcinoma cells, spleen cells, macrophage, and RAW264.7 cells. To investigate innate immunomodulating effects of LPE on macrophage, we measured tumor necrosis factor-alpha (TNF-α), interleukin-12 (IL-12), and interleukin-10 (IL-10). To investigate innate immunomodulating effects of LPE on RAW264.7 cell, we measured TNF-α, interleukin-6 (IL-6). In addition, TNF-α and nitric oxide (NO) induced by lipopolysaccharide (LPS) were measured after treating with LPE to observe innate immunomodulating effect of LPE on RAW264.7 cell. Also, mitogen-activated protein kinase (MAPK) and nuclear factor κB (NF-κB) were examined by western blot analysis. Results: In an in vitro cytotoxicity analysis, LPE affected tumor cell growth above specific concentration. As compared with the control group, the production of TNF-α, IL-12, and IL-10 were increased in macrophage. As compared with the control group, the production of TNF-α and IL-6 were increased in RAW 264.7 cell. The expression of TNF-α and NO induced by LPS after treating LPE was decreased. In addition, treatment of RAW 264.7 cell with LPE increased the phosphorylation levels of p-extracellular signal-regulated kinase (p-ERK), p-Jun N-terminal kinase (p-JNK), and p-p38. Conclusions: LPE might have impact on the anti-cancer effect by activation of innate immune system and inflammation control.