• 생명과학회지
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• 제23권4호
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• pp.471-478
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• 2013

인체 중간엽 줄기세포는 다양한 세포로의 분화 및 자가증식 할 수 있는 능력뿐만 아니라 질병치료에 대한 치료적 잠재력을 가지고 있다. 줄기세포 분화의 분자 기작에 대한 이해는 줄기세포 이식의 치료 효능을 향상시킨다. 본 연구에는 인체 중간엽 줄기세포의 골분화에서 NFAT5의 역할을 밝혔다. 특이적 siRNA의 transfection으로 인한 NFAT5의 억제는 인체 중간엽 줄기세포의 골분화를 현저히 감소시켰으며, NF-${\kappa}B$ promoter 활성화 또한 세포의 증식이나 지방 세포로의 분화에 영향 없이 감소 시켰다. NFAT5의 발현 억제는 기본적으로 유도되는 NF-${\kappa}B$의 활성화와 TNF-${\alpha}$에 의해서 유도되는 NF-${\kappa}B$의 활성화를 감소시켰으나, TNF-${\alpha}$에 의해서 유도되는 NF-${\kappa}B$의 분해에는 아무런 영향을 주지 않았다. 이번 연구를 통해 NFAT5가 NF-${\kappa}B$ 경로를 조절함으로써 인체 중간엽 줄기 세포의 골분화에 아주 중요한 역할을 하는 것을 확인 할 수 있었다.

• 대한본초학회지
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• 제33권5호
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• pp.73-79
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• 2018

Objectives : Coriandrum sativum is a medicinal herb that is used to enhance organoleptic quality and food flavor and as source of natural antioxidants. This research investigated the anti-inflammatory activity of Coriandrum sativum stem methanol extract (CSSE) using RAW 264.7 cells. Methods : Production of tumor necrosis factor-${\alpha}$(TNF-${\alpha}$), interleukin (IL)-$1{\beta}$, IL-6, and nitric oxide (NO) in the culture supernatant, protein expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and nuclear factor-kappa B (NF-${\kappa}B$) in the extract were assayed. Results : Treatment with CSSE ($100{\mu}g/m{\ell}$) resulted in inhibited levels of protein expression of lipopolysaccharide- (LPS-) induced iNOS, COX-2, and NF-${\kappa}B$ as well as production of TNF-${\alpha}$, IL-$1{\beta}$, IL-6, and NO induced by LPS. Conclusions : These results demonstrate that CSSE exhibits anti-inflammatory activities via decreasing production of pro-inflammatory mediators through suppression of the pathways of NF-${\kappa}B$ in LPS-induced RAW 264.7 cells. Thus, CSSE may have therapeutic potential for a variety of inflammation-mediated diseases.

• Biomolecules & Therapeutics
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• 제21권6호
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• pp.435-441
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• 2013

Emodin, a naturally occurring anthraquinone derivative isolated from Polygoni cuspidati radix, has several beneficial pharmacologic effects, which include anti-cancer, anti-diabetic, and anti-inflammatory activities. In this study, the authors examined the effect of emodin on the production of proinflammatory cytokines, such as, tumor necrosis factor (TNF)-${\alpha}$ and interleukin (IL)-6, in mouse bone marrow-derived mast cells (BMMCs) stimulated with phorbol 12-myristate 13-acetate (PMA) plus the calcium ionophore A23187. To investigate the mechanism responsible for the regulation of pro-inflammatory cytokine production by emodin, the authors assessed its effects on the activations of transcriptional factor nuclear factor-${\kappa}B$ (NF-${\kappa}B$) and mitogen-activated protein kinases (MAPKs). Emodin attenuated the nuclear translocation of (NF)-${\kappa}B$ p65 and its DNA-binding activity by reducing the phosphorylation and degradation of $I{\kappa}B{\alpha}$ and the phosphorylation of $I{\kappa}B$ kinase B (IKK). Furthermore, emodin dose-dependently attenuated the phosphorylations of MAPKs, such as, extracellular signal-regulated kinase 1/2 (ERK1/2), p38 MAP kinase, and the stress-activated protein kinases (SAPK)/c-Jun-N-terminal kinase (JNK). Taken together, the findings of this study suggest that the anti-inflammatory effects of emodin on PMA plus A23187-stimulated BMMCs are mediated via the inhibition of NF-${\kappa}B$ activation and of the MAPK pathway.

• 한방안이비인후피부과학회지
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• 제26권4호
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• pp.70-80
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• 2013

Objectives : Haliotidis Concha has been used to treat various human diseases such as liver dysfunction and inflammatory disorder. Although it has been shown the effects of Haliotidis Concha on the various diseases, it has almost not been studied about the anti-inflammatory effects of the Haliotidis Concha and its mechanisms. Methods : This research investigated the effects of the Haliotidis Concha ethanol extract (HCE) on the production of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) as well as tumor necrosis factor-alpha (TNF-${\alpha}$). The protein expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were assayed by immunoblot analyses, and the productions of NO, $PGE_2$ and TNF-${\alpha}$ were assessed by ELISA. Results : Haliotidis Concha decreased the production of NO and $PGE_2$, and inhibited the expression iNOS and COX-2 proteins in a concentration-dependent manner in LPS-treated Raw 264.7 cells. HCE suppressed the ability of LPS to activate the signaling pathways of nuclear factor kappa B (NF-${\kappa}B$) as indicated by HCE inhibited nuclear NF-${\kappa}B$ level and I-${\kappa}B{\alpha}$ phosphorylation. Also, HCE inhibited mitogen-activated protein kinases (MAPKs). Conclusions : HCE repressed the production of LPS-inducible NO, $PGE_2$ and TNF-${\alpha}$, which may be mediated by inhibition of NF-${\kappa}B$ translocation. This study suggest the use for the treatment of acute inflammatory disorders.

• Journal of Acupuncture Research
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• 제24권2호
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• pp.15-29
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• 2007

목적 : 후박(厚朴)(Magnolia Obovata)에서 추출한 낮은 농도의 Obovatol 약침액의 RAW264.7 세포에서 LPS로 유발된 염증，$TNF-{\alpha}$로 유발된 human Prostate carcinoma PC-3 및 LNCap 세포의 세포증식에 대한 영향과 그 기전을 살펴보고자 하였다. 방법 : RAW264.7 세포에서 LPS로 염증을 유발하고 낮은 농도의 Obovatol 약침액을 처리한 후 cell viability, NO 생성량，iNOS와 COX-2의 발현, $NF-{\kappa}B$활성，전사능력을 관찰하기 위해 MTT assay, NO determination assay, western blot analysis, EMSA, luciferase activity assay를 시행하였고，LNCap, PC-3 세포에 $TNF-{\alpha}$로 증식을 유도하고 낮은 농도의 Obovatol 약침액을 처리한 후 cell growth, apoptosis 및 apoptosis와 연관된 $NF-{\kappa}B$의 활성 변화를 관찰하기 위해 WST-1, Cell morphogy test, DAPI staining and TUNEL assay, EMSA, luciferase activity assay를 시행하였다. 결과 : 1. RAW264.7 세포에서 낮은 농도의 Obovatol 약침액 처리는 $NF-{\kappa}B$의 활성 및 전사능력을 낮추고 iNOS와 COX-2의 발현과 NO 생성을 감소시켜 LPS로 유발된 염증을 억제하였다. 2. LNCap, PC-3 세포에서 낮은 농도의 Obovatol 약침액 처리는 $NF-{\kappa}B$의 활성을 낮추어 세포자별사를 촉진함으로써 $TNF-{\alpha}$로 유발된 암세포의 성장을 억제하였다. 결론 : 이상의 결과는 낮은 농도의 Obovatol 약침액이 항염 및 인간 전립선암세포주인 PC-3, LNCap에 대한 증식억제 효과가 있음을 입증한 것이며，향후 이를 바탕으로한 생체 연구에서의 긍정적인 결과는 Obovatol 약침액이 만성염증성 질환 및 전립선암의 예방과 치료에 대한 효과적인 치료제 개발에 초석이 될 것으로 기대된다.

• 대한한방부인과학회지
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• 제21권2호
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• pp.166-183
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• 2008

Purpose: This study was performed to determine anti-imflammatory effects of Haedoksamultang. Methods: In this study, I examined the effects of Haedoksamultang on the production of nitric oxide(NO), tumor necrosis factor-${\alpha}$(TNF-${\alpha}$), and interlukin-1${\beta}$(IL-1${\beta}$) as well as the expression of inducible NO synthase(iNOS), cyclooxygenase-2(COX-2), TNF-${\alpha}$, and IL-1${\beta}$ in RAW 264.7 cells. Haedoksamultang inhibited LPS-stimulated NO production. Western blotting and RT-PCR analysis showed that Haedoksamultang suppressed LPS-induced iNOS and COX-2 protein and mRNA expression in RAW 264.7 cells. Haedoksamultang also suppressed the expression and production of LPS-stimulated TNF-${\alpha}$ and IL-1${\beta}$ in RAW 264.7 cells. Haedoksamultang inhibited NF-${\kappa}B$ activation in LPS-treated RAW 264.7 cells. Moreover, this compound blocked $I{\kappa}B-{\alpha}$ phosphorylation and nuclear translocation of the cytosolic NF-${\kappa}B$ p65 subunit, which highly correlated with the production and expression of inflammatory mediators. Results: Haedoksamultang suppresses that inflammation-associated gene expression by blocking NF-${\kappa}B$ activation. Conclusion: These results suggest that Haedoksamultang may be beneficial for treating inflammatory disease.

• 한방안이비인후피부과학회지
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• 제25권3호
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• pp.1-12
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• 2012

Objectives : Fagopyrum esculentum(FE) has been used for removal of inflammation of internal organs and treatment of sore and ulcer by heat toxin in Korean herbal medicines. In this study, To investigated the protective effect of FE on allergic response, we determined whether FE inhibits allergic response. Methods : The effect of FE was analyzed by ELISA, RT-PCR and Western blot in RBL-2H3 cells. We investigated cell viability, ${\beta}$-hexosaminidase, as a marker of degranulation, cytokne, and intracellular ROS and MAPK and NF-${\kappa}B$ signaling. Results : We found that FE suppressed ${\beta}$-hexosaminidase release, the production of IL-4 and TNF-${\alpha}$ and intracellular ROS level in RBL-2H3 by the anti-DNP IgE plus DNP-HSA stimulation. FE also significantly inhibited cytokine mRNA expressions, such as IL-$1{\beta}$, IL-2, IL-3, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ and GM-CSF in RBL-2H3. In addition, PF suppressed the phospholyation of ERK1/2, JNK1/2, p38 and $I{\kappa}B{\alpha}$ and NF-${\kappa}B$ signal transduction pathway. Conclusions : Our results indicate that FE protects against allergic response and exerts an anti-inflammatory effect through the inhibition of degranulation and production of cytokines and ROS via the suppression MAPK and NF-${\kappa}B$ of signal transduction. Abbrevations : FE, Fagopyrum esculentum; RBL-2H3, rat basophilic leukemia cell line; ROS, reactive oxygen species; MAPK, Mitogen-activated protein kinase; $NF{\kappa}B$, nuclear factor ${\kappa}B$; $TNF{\alpha}$, Tumor necrosis factor alpha; GM-CSF, Granulocyte macrophage colony-stimulating factor; ERK, extracellular-signal-regulated kinase; JNK, c-Jun NH2-terminal kinase; p38, p38 MAP kinase; $I{\kappa}B{\alpha}$, inhibitory-kappa B alpha.

• 한국버섯학회지
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• 제11권1호
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• pp.46-51
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• 2013

꽃송이버섯 추출물은 염증반응 시 유도되는 NO 생성을 저해하는 활성이 있는 것으로 나타났으며, 200 ${\mu}g/ml$의 꽃송이버섯 추출물을 처리하였을 때 NO 저해능이 최대 효과를 보였고 RAW264.7 cell에서는 대조군과 유사한 수준으로 NO 생성을 억제하였다. 이러한 NO 생성의 저해는 iNOS의 발현이 감소한 것에 의한 결과임을 단백질과 mRNA의 발현량 변화를 통하여 확인하였으며, mRNA 발현 변화는 iNOS 유전자의 전사를 담당하는 전사인자인 $NF-{\kappa}B$와 STAT-1의 활성감소에 의한 결과임을 western blot을 통하여 확인하였다. 특히, $NF-{\kappa}B$의 활성감소는 $I{\kappa}B{\alpha}$의 활성증가에 의한 $NF-{\kappa}B$의 억제능 향상에 의한 것임을 확인하였고, 활성억제된 $NF-{\kappa}B$가 핵 내부로의 이동이 저해되면서 iNOS 유전자 발현에 영향을 미친 것임을 확인하였다. 그러므로, 꽃송이버섯 추출물은 NO 저해능을 이용한 항염증소재로서 염증성질환의 완하에 도움이 될 것으로 사료된다.

• 동의생리병리학회지
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• 제23권3호
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• pp.673-677
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• 2009

In order to verify the anti-inflammatory effects of Gelidium amansii, RAW264.7 macrophages were incubated with the extract of 70% ethanol solution (Ex), and activated with the endotoxin lipopolysaccharide (LPS). Ex inhibited the expression of the pro-inflammatory enzymes, including inducible nitric oxide (NO) synthase (iNOS) and cyclooxygenase-2 (COX-2), and the production of iNOS-mediated NO and COX-2-mediated prostglandin $E_2$ ($PGE_2$) production in a dose-dependent manner. Ex also reduced the release of the pro-inflammatory cytokines, including tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-1${\beta}$ (IL-1${\beta}$) and IL-6 in LPS-activated macrophages, The observed anti-inflammatory effects of Ex was associated with inactivation of the nuclear factor ${\kappa}B$ (NF-${\kappa}B$) that mediates the induction of iNOS, COX-2, TNF-${\alpha}$, IL-1${\beta}$, and IL-6. Further studies showed that Ex inactivated NF-${\kappa}B$ through inhibition of phosphorylation of the inhibitory ${\kappa}B$ ($l{\kappa}B$), Taken together, these results suggest that Gelidium amansii exerts anti-inflammatory effects by inhibiting the expression of pro-inflammatory enzymes and the secretion of pro-inflammatory cytokines via inactivation of NF-${\kappa}B$ and/or $l{\kappa}B$.

• Archives of Pharmacal Research
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• 제27권9호
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• pp.954-960
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• 2004

Expression of the NF-$textsc{k}$B-dependent genes responsible for inflammation, such as TNF-$\alpha$, IL-1$\beta$, and nitric oxide synthase (NOS), contributes to chronic inflammation which is a major cause of neurodegenerative diseases (i.e. Alzheimer＇s disease). Although NF-$textsc{k}$B plays a biphasic role in different cells like neurons and microglia, controlling the activation of NF-$textsc{k}$B is important for its negative feedback in either activation or inactivation. In this study, we found that ursodeoxycholic acid (UDCA) inhibited I$textsc{k}$B$\alpha$ degradation to block expression of the NF-$textsc{k}$B-dependent genes in microglia when activated by $\beta$-amyloid peptide (A$\beta$). We also showed that when microglia is activated by $A\beta$42, the expression of A20 is suppressed. These findings place A20 in the category of ' protective ' genes, protecting cells from pro-inflammatory reper-toires induced in response to inflammatory stimuli in activated microglia via NF-$textsc{k}$B activation. In light of the gene and proteins for NF-$textsc{k}$B-dependent gene and inactivator for NF-$textsc{k}$B (I$textsc{k}$B$\alpha$), the observations now reported suggest that UDCA plays a role in supporting the attenuation of the production of pro-inflammatory cytokines and NO via inactivation of NF-$textsc{k}$B. Moreover, an NF-$textsc{k}$B inhibitor such as A20 can collaborate and at least enhance the anti-inflammatory effect in microglia, thus giving a potent benefit for the treatment of neurodegenerative diseases such as AD.uch as AD.