• Journal of Ginseng Research
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• 제46권4호
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• pp.550-560
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• 2022

Background: The effect of ginsenoside Rh2 (G-Rh2) on mast cell-mediated anaphylaxis remains unclear. Herein, we investigated the effects of G-Rh2 on OVA-induced asthmatic mice and on mast cell-mediated anaphylaxis. Methods: Asthma model was established for evaluating airway changes and ear allergy. RPMCs and RBL-2H3 were used for in vitro experiments. Calcium uptake, histamine release and degranulation were detected. ELISA and Western blot measured cytokine and protein levels, respectively. Results: G-Rh2 inhibited OVA-induced airway remodeling, the production of TNF-α, IL-4, IL-8, IL-1β and the degranulation of mast cells of asthmatic mice. G-Rh2 inhibited the activation of Syk and Lyn in lung tissue of OVA-induced asthmatic mice. G-Rh2 inhibited serum IgE production in OVA induced asthmatic mice. Furthermore, G-Rh2 reduced the ear allergy in IgE-sensitized mice. G-Rh2 decreased the ear thickness. In vitro experiments G-Rh2 significantly reduced calcium uptake and inhibited histamine release and degranulation in RPMCs. In addition, G-Rh2 reduced the production of IL-1β, TNF-α, IL-8, and IL-4 in IgE-sensitized RBL-2H3 cells. Interestingly, G-Rh2 was involved in the FcεRI pathway activation of mast cells and the transduction of the Lyn/Syk signaling pathway. G-Rh2 inhibited PI3K activity in a dose-dependent manner. By blocking the antigen-induced phosphorylation of Lyn, Syk, LAT, PLCγ2, PI3K ERK1/2 and Raf-1 expression, G-Rh2 inhibited the NF-κB, AKT-Nrf2, and p38MAPK-Nrf2 pathways. However, G-Rh2 up-regulated Keap-1 expression. Meanwhile, G-Rh2 reduced the levels of p-AKT, p38MAPK and Nrf2 in RBL-2H3 sensitized IgE cells and inhibited NF-κB signaling pathway activation by activating the AKT-Nrf2 and p38MAPK-Nrf2 pathways. Conclusion: G-Rh2 inhibits mast cell-induced allergic inflammation, which might be mediated by the AKT-Nrf2/NF-kB and p38MAPK-Nrf2/NF-κB signaling pathways.

• 대한본초학회지
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• 제37권6호
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• pp.19-28
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• 2022

Objectives : To develop natural ingredients that help prevent or treat anti-inflammatory-related diseases and use themas basic data, we investigated anti-inflammatory activity of Cynanchi Atrati Radix Et Rhizoma water extracts(CWE) in lipopolysaccharide(LPS)-induced murine macrophage cell line, RAW 264.7 cells. Methods : The cell viabilities were evaluated with RAW 264.7 cells. The production of nitric oxide(NO), prostaglandin E2(PGE2), pro-inflammatory cytokines such tumor necrotic factor(TNF)-α and interleukin(IL)-6 were assessed in LPS-induced RAW 264.7 cell treated with CWE. Furthermore, the protein expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2(COX-2), and mitogen-activated protein kinase(MAPK) were assessed by western blotting. Results : In RAW 264.7 cell, the cell viability by CWE treatment was more than 98.4% at a concentration of 100-400 ㎍/mL. At a concentration of 800 ug/ml of CWE, the cell viability was as low as 86%. At doses of 100, 200 and 400 ㎍/mL, CWE inhibited the production of NO, PGE2, TNF-𝛼 and IL-6 in a dose-dependent manner and also decreased the expression of iNOS and COX-2 from LPS-induced RAW 264.7 cells. In addition, CWE significantly inhibited the MAPK pathway including decreased the phosphorylation of the p38, c-Jun N-terminal kinase(JNK) and extracellular signal-regulated kinase(ERK1/2). Conclusions : Our study provides evidence that CWE inhibits the production of main pro-inflammatory molecules in LPS-induced RAW 264.7 cells via expression of p38, JNK, and ERK1/2 MAPK signaling pathways. Therefore, CWE is expected to be widely used as a natural ingredient for anti-inflammatory functional foods or pharmaceuticals in the future.

• Tuberculosis and Respiratory Diseases
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• 제58권6호
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• pp.590-599
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• 2005

연구배경 : 만성폐쇄성폐질환에서 나타나는 기도점액의 과다분비는 이 질환의 중요한 병리학적 소견이며 호흡곤란 등 환자의 증상을 악화시키는 요인 중의 하나이다. 기도 점액을 구성하는 여러 성분 중 Muc 유전자에 의해 만들어지는 당 단백이 흡연에 의해 생성이 증가하는데 이에 관여하는 세포 내 신호전달 과정에 대하여 확실히 밝혀진 바가 없다. 저자는 Muc 유전자 중 인체의 기도에 가장 많이 분비되는 Muc5ac 점액 생성을 담당하는 Muc5ac 유전자의 발현이 흡연에 의하여 증가하는데 관여하는 세포 내 신호전달 과정을 알아보고자 하였다. 재료 및 방법 : 사람 폐선암 세포주인 A549 세포를 배양하여 Muc5ac 유전자의 promotor를 luciferase reporter plasmid를 사용하여 세포 내에 transfection시키고 5% 담배연기 추출물로 자극하여 배양하였다. 또 세포 내 신호전달에 관여하는 표피성장인자 수용체 kinase의 억제제인 AG1478, mitogen-activated protein kinase kinase(MAPKK) 억제제인 PD98059, p38 mitogen-activated protein kinase 억제제인 SB203580으로 각각 전 처치 후 역시 5% 담배연기 추출물로 자극 배양하였다. 배양된 세포에서 단백질을 추출하여 luciferase 분석을 통하여 Muc5ac promoter 활성도를 측정하고 Western blot을 이용하여 표피성장인자 수용체와 mitogen-activated protein kinase (MAPK)인 extracellular signalrelated kinase (ERK)1/2, p38 MAPK, c-Jun N-terminal kinase (JNK)의 발현을 확인하였다. 또 세포에서 RNA를 추출한 후 Muc5ac primer를 이용하여 역전사효소 중합연쇄반응을 수행하여 Muc5ac mRNA 발현을 관찰하였다. 결 과 : 1. Muc5ac promoter를 삽입한 A549 세포를 5% 담배연기 추출물로 자극하였을 때 의의 있게 luciferase 활성도가 증가하였고(P<0.001) 자극하는 시간이 3시간이었을 때 luciferase 활성도가 최고치를 보였다(P<0.01). 또 담배연기 추출물 자극은 표피성장인자 수용체를 인산화시켰으며 인산화는 AG1478과 PD98059에 의하여 억제되었다. 2. AG1478 혹은 PD98059로 전 처치 후 5% 담배연기 추출물로 자극한 경우 5% 담배연기 추출물 단독으로 자극한 것에 비하여 유의하게 lucifearse 활성도가 억제되었고 (P<0.01) 세 가지 종류의 MAPK 중 ERK1/2와 p38 MAPK의 인산화는 관찰되었으나 JNK의 인산화는 관찰되지 않았다. 역전사효소 중합연쇄반응을 이용하여 관찰한 Muc5ac mRNA 발현은 담배연기 추출물에 의해 증가되었고 PD98059와 AG1478에 의하여 역시 억제되었다. 3. 담배연기 추출물에 의하여 인산화 된 ERK1/2는 PD98059에 의하여 인산화가 감소하였고 p38 MAPK의 인산화는 PD98059와 SB203580에 의하여 감소하였으며 이 두 가지 억제제는 모두 luciferase 활성도를 유의하게 억제시켰다(P<0.0001). 결 론 : 담배연기 추출물은 Muc5ac 유전자의 발현을 증가시켜 기도 내 점액 분비를 증가시키며 이는 표피성장인자 수용체를 매개로 ERK1/2와 p38 MAPK를 경유하여 세포 내 신호전달이 이루어진다고 생각된다. 따라서 점액 유전자 활성화를 매개하는 신호전달 과정을 차단하는 약제나 방법이 개발된다면 과도한 점액분비를 치료할 수 있을 것으로 생각한다.

• The Korean Journal of Physiology and Pharmacology
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• 제9권5호
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• pp.299-304
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• 2005

Cardiac hypertrophy contributes an increased risk to major cerebrovascular events. However, the molecular mechanisms underlying cerebrovascular dysfunction during cardiac hypertrophy have not yet been characterized. In the present study, we examined the molecular mechanism of isoproterenol (ISO)-evoked activation of Ras/Raf/MAPK pathways as well as PKA activity in cerebral artery of rabbits, and we also studied whether the activations of these signaling pathways were altered in cerebral artery, during ISO-induced cardiac hypertrophy compared to heart itself. The results show that the mRNA level of c-fos (not c-jun and c-myc) in heart and these genes in cerebral artery were considerably increased during cardiac hypertrophy. These results that the PKA activity and activations of Ras/Raf/ERK cascade as well as c-fos expression in rabbit heart during cardiac hypertrophy were consistent with previous reports. Interestingly, however, we also showed a novel finding that the decreased PKA activity might have differential effects on Ras and Raf expression in cerebral artery during cardiac hypertrophy. In conclusion, there are differences in molecular mechanisms between heart and cerebral artery during cardiac hypertrophy when stimulated with β2 adrenoreceptor (AR), suggesting a possible mechanism underlying cerebrovascular dysfunction during cardiac hypertrophy.

• 생명과학회지
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• 제32권11호
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• pp.899-907
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• 2022

Quercetin은 과일과 채소에 풍부한 플라보노이드 중의 하나로써, 항산화, 항염증, 항암, 항바이러스 활성 등 다양한 약리학적 활성을 가지고 있는 것으로 알려져 있다. 본 연구에서는 in vitro 모델에서 quercetin의 항염증 활성과 작용기전을 연구하였다. Quercetin은 LPS로 자극된 RAW264.7에서 세포 생존율에 영향 없이 NO 생산을 농도 의존적으로 저해하였고, iNOS와 COX-2 단백질의 발현을 억제하였다. 게다가, quercetin은 LPS로 유도된 p38, JNK, ERK의 인산화를 농도 의존적으로 저해하였고, NF-κB p65 단백질과 억제자인 IκBα 단백질의 인산화를 저해하였다. 이러한 결과는 quercetin의 항염증 활성이 MAPK 경로와 NF-κB를 조절함으로써 이루어진다는 것을 시사한다. Quercetin에 의해 4종류의 친 염증성 cytokine (CSF2, IL-1β, IL-6, TNF-α)의 발현 변화를 정량적 real-time PCR 방법으로 확인한 결과, 모든 cytokine 유전자의 발현이 감소됨을 확인하였다. 종합적으로, 본 연구결과는 플라보노이드 quercetin이 RAW264.7 세포에서 LPS로 유도된 염증반응을 MAPK 경로와 NF-κB경로를 통해 억제하고 친염증성 cytokine 유전자의 발현을 억제함으 로써 조절한다는 것을 제시한다.

• BMB Reports
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• 제46권7호
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• pp.352-357
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• 2013

Atherosclerosis, which manifests as acute coronary syndrome, stroke, and peripheral arterial diseases, is a chronic inflammatory disease of the arterial wall. Prunella vulgaris, a perennial herb with a worldwide distribution, has been used as a traditional medicine in inflammatory disease. Here, we investigated the effects of P. vulgaris ethanol extract on TNF-${\alpha}$-induced inflammatory responses in human aortic smooth muscle cells (HASMCs). We found that P. vulgaris ethanol extract inhibited adhesion of monocyte/macrophage-like THP-1 cells to activated HASMCs. It also decreased expression of intercellular adhesion molecule-1, vascular cell adhesion molecule-1, E-selectin and ROS, No production in TNF-${\alpha}$-induced HASMCs and reduced NF-${\kappa}B$ activation. Furthermore, P. vulgaris extract suppressed TNF-${\alpha}$-induced phosphorylation of p38 mitogen-activated protein kinase (MAPK) and extracellular signal-regulated kinase (ERK). These results demonstrate that P. vulgaris possesses anti-inflammatory properties and can regulate TNF-${\alpha}$-induced expression of adhesion molecules by inhibiting the p38 MAPK/ERK signaling pathway.

• Journal of Microbiology and Biotechnology
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• 제31권5호
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• pp.726-732
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• 2021

In this study, we evaluated the immune-enhancing activity of kimchi-derived Lactobacillus plantarum 200655 on immune suppression by cyclophosphamide (CP) in ICR mice. Animals were fed distilled water or 1×10⁹ colony-forming unit/kg B.W. 200655 or Lactobacillus rhamnosus GG as a positive control for 14 days. An in vivo model of immunosuppression was induced using CP 150 and 100 mg/kg B.W. at 7 and 10 days, respectively. Body weight, spleen index, spleen weight, and gene expression were measured to estimate the immune-enhancing effects. The dead 200655 (D-200655) group showed an increased spleen weight compared to the sham control (SC) group. Similarly, the spleen index was significantly higher than that in the CP-treated group. The live 200655 (L-200655) group showed an increased mRNA expression of tumor necrosis factor-alpha (TNF-α) and interleukin (IL)-6 in splenocytes. Also, the iNOS and COX-2 mRNA expression was upregulated in the L-200655 group compared to the CP-only (SC) group. The phosphorylation of ERK and MAPK was also upmodulated in the L-200655 group. These results indicate that L. plantarum 200655 ameliorated CP-induced immune suppression, suggesting that L. plantarum 200655 may have the potential to enhance the immune system.

• 한국융합학회논문지
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• 제13권1호
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• pp.101-107
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• 2022

Tertomotide는 항암제로 개발된 펩타이드 백신이다. 그러나 동물실험과 임상시험에서 염증성 증상이 완화되는 현상이 발견된바 있다. 이에 tertomotide가 항염물질로 작용하는지 확인하기 위하여 직접적인 항염활성과 그 작용기전을 조사하였다. 이를 위해 LPS 또는 PMA에 의해 활성화된 monocyte에 tertomotide를 처리한 후 염증성 cytokine 생산과 관련된 신호전달과정을 관찰하였다. Monocyte에서 tertomotide는 TNF-α, IL-1β, IL-8 등 염증성 싸이토카인의 생산을 감소시켰고 NF-κB 신호를 감쇄시켰으며 또한 TNF-α에 의한 ERK1/2와 P38 MAPK의 활성화를 저해하였다. 이 결과는 tertomotide 처치에 따른 염증성 질환 완화가 NF-κB/STAT3의 신호의 감쇄와 항염활성에 의한 것이라고 설명할 수 있고 이를 활용하여 신규 항염 약물의 도출이 가능할 것으로 판단된다. 이는 면역학적 활성을 목표로 계산화학적으로 설계된 물질의 생물학적 성질을 활용하여 새로운 약물을 도출하는 융합연구의 예시가 될 것이다.

• 한국자원식물학회지
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• 제31권6호
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• pp.634-640
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• 2018

Aralia cordata (A. cordata), which belongs to Araliaceae, is a perennial herb widely distributed in East Asia. We evaluated the anti-inflammatory effect of stems (AC-S), roots (AC-R) and leaves (AC-L) extracted with 100% methanol of A. cordata and elucidated the potential signaling pathway in LPS-stimulated RAW264.7 cells. The AC-L showed a strong anti-inflammatory activity through inhibition of NO production. AC-L dose-dependently inhibited NO production by suppressing iNOS, COX-2 and $IL-{\beta}$ expression in LPS-stimulated RAW264.7 cells. AC-L inhibited the degradation and phosphorylation of $I{\kappa}B-{\alpha}$, which donated to the inhibition of p65 nuclear accumulation and $NF-{\kappa}B$ activation. Furthermore, AC-L suppressed the phosphorylation of ERK1/2 and p38. These results suggested that AC-L may utilize anti-inflammatory activity by blocking $NF-{\kappa}B$ and MAPK signaling pathway and indicated that the AC-L can be used as a natural anti-inflammatory drugs.

• Archives of Pharmacal Research
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• 제23권1호
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• pp.1-16
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• 2000

Many natural products elicit diverse pharmacological effects. Using two classes of potential chemopreventive compounds, the phenolic compounds and the isothiocyanates, we review the potential utility of two signaling events, the mitogen-activated protein kinases (MAPKs) and the ICE/Ced-3 proteases (caspases) stimulated by these agents in mammalian cell lines. Studies with phenolic antioxidants (BHA, tBHQ), and natural products (flavonoids; EGCG, ECG, and isothiocyanates; PEITC, sulforaphane), provided important insights into the signaling pathways induced by these compounds. At low concentrations, these chemicals may activate the MAPK (ERK2, JNK1, p38) leading to gene expression of survival genes (c-Fos, c-Jun) and defensive genes (Phase II detoxifying enzymes; GST, QR) resulting in survival and protective mechanisms (homeostasis response). Increasing the concentrations of these compounds will additionally activate the caspase pathway, leading to apoptosis (potential cytotoxicity). Further increment to suprapharmacological concentrations will lead to nonspecific necrotic cell death. The wider and narrow concentration ranges between the activation of MAPK/gene induction and caspases/cell death exhibited by phenolic compounds and isothiocyanates, respectively, in mammalian cells, may reflect their respective therapeutic windows in vivo. Consequently, the studies of signaling pathways elicited by natural products will advance our understanding of their efficacy and safety, of which many man become important therapeuitc drugs of the future.