• Biomolecules & Therapeutics
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• 제31권2호
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• pp.200-209
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• 2023

Patients with non-small-cell lung cancer (NSCLC) with epidermal growth factor receptor (EGFR) amplification or sensitive mutations initially respond to the tyrosine kinase inhibitor gefitinib, however, the treatment becomes less effective over time by resistance mechanism including mesenchymal-epithelial transition (MET) overexpression. A therapeutic strategy targeting MET and EGFR may be a means to overcoming resistance to gefitinib. In the present study, we found that picropodophyllotoxin (PPT), derived from the roots of Podophyllum hexandrum, inhibited both EGFR and MET in NSCLC cells. The antitumor efficacy of PPT in gefitinib-resistant NSCLC cells (HCC827GR), was confirmed by suppression of cell proliferation and anchorage-independent colony growth. In the targeting of EGFR and MET, PPT bound with EGFR and MET, ex vivo, and blocked both kinases activity. The binding sites between PPT and EGFR or MET in the computational docking model were predicted at Gly772/Met769 and Arg1086/Tyr1230 of each ATP-binding pocket, respectively. PPT treatment of HCC827GR cells increased the number of annexin V-positive and subG1 cells. PPT also caused G2/M cell-cycle arrest together with related protein regulation. The inhibition of EGFR and MET by PPT treatment led to decreases in the phosphorylation of the downstream-proteins, AKT and ERK. In addition, PPT induced reactive oxygen species (ROS) production and GRP78, CHOP, DR5, and DR4 expression, mitochondrial dysfunction, and regulated involving signal-proteins. Taken together, PPT alleviated gefitinib-resistant NSCLC cell growth and induced apoptosis by reducing EGFR and MET activity. Therefore, our results suggest that PPT can be a promising therapeutic agent for gefitinib-resistant NSCLC.

• Biomolecules & Therapeutics
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• 제22권2호
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• pp.129-135
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• 2014

Previously, we reported that lysophosphatidylethanolamine (LPE), a lyso-type metabolite of phosphatidylethanolamine, can increase intracellular $Ca^{2+}$ ($[Ca^{2+}]_i$) via type 1 lysophosphatidic acid (LPA) receptor ($LPA_1$) and CD97, an adhesion G-protein-coupled receptor (GPCR), in MDA-MB-231 breast cancer cells. Furthermore, LPE signaling was suggested as like $LPA_1/CD97-G_{i/o}$ proteins-phospholipase $C-IP_3-Ca^{2+}$ increase in these cells. In the present study, we further investigated actions of LPE not only in the $[Ca^{2+}]_i$ increasing effect but also in cell proliferation and migration in MDA-MB-231 breast cancer cells. We utilized chemically different LPEs and a specific inhibitor of $LPA_1$, AM-095 in comparison with responses in SK-OV3 ovarian cancer cells. It was found that LPE-induced $Ca^{2+}$ response in MDA-MB-231 cells was evoked in a different manner to that in SK-OV3 cells in terms of structural requirements. AM-095 inhibited LPE-induced $Ca^{2+}$ response and cell proliferation in MDA-MB-231 cells, but not in SK-OV3 cells, supporting $LPA_1$ involvement only in MDA-MB-231 cells. LPA had significant effects on cell proliferation and migration in MDA-MB-231 cells, whereas LPE had less or no significant effect. However, LPE modulations of MAPKs (ERK1/2, JNK and p38 MAPK) was not different to those by LPA in the cells. These data support the involvement of LPA1 in LPE-induced $Ca^{2+}$ response and cell proliferation in breast MDA-MB-231 cells but unknown GPCRs (not $LPA_1$) in LPE-induced responses in SK-OV3 cells. Furthermore, although LPE and LPA utilized $LPA_1$, LPA utilized more signaling cascades than LPE, resulting in stronger responses by LPA in proliferation and migration than LPE in MDA-MB-231 cells.

• 대한화장품학회지
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• 제30권2호
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• pp.247-251
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• 2004

자외선은 피부에 염증반응이나 광노화와 같은 다양한 반응을 야기시킨다고 알려져 있다. 특히 자외선에 의해 손상을 받은 피부는 콜라겐의 양이 감소되어 있는데, 이는 자외선에 의해 피부 내에서 콜라겐을 분해하는 효소(MMP, matrix metalloproteinases)의 양이 증가하기 때문이라고 알려져 왔다. 또한 자외선에 의해 피부에서 염증반응이 유발되는데, 이러한 반응은 프로스타글란딘이라는 물질에 의해 매개되며, 이 프로스타글란딘에 의해서도 MMP가 증가한다고 알려져 왔다. 본 연구에서는 6개월의 임상실험을 통해 광노화된 피부에서 주름형성억제효능이 뛰어난 FDP(fructose 1.6-diphosphate)의 작용기전을 인체각질형성 세포를 이용하여 연구하였다. 인체각질형성세포에 자외선을 조사할 경우 프로스타글란딘, COX-2(cyclooxygenase-2), MMPs의 활성이 증가하는 것을 확인하였며, 이는 FDP의 처리에 의해 감소되었다. 이러한 효과는 자외선에 의해 인체각질형성세포에서 발생하는 신호전달과정을 억제함으로써 일어나는 효과임이 증명되었다. 따라서, FDP는 자외선에 의해 일어나는 세포 내 신호전달과정을 억제하며, 이로 인해 야기되는 프로스타글란딘, COX-2, MMPs의 증가를 억제함으로써 피부의 광노화를 억제할 수 있는 원료로 여겨진다.

• Archives of Pharmacal Research
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• 제29권10호
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• pp.911-920
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• 2006

Phenolic antioxidant butylated hydroxyanisole (BHA) is a commonly used food preservative with broad biological activities, including protection against chemical-induced carcinogenesis, acute toxicity of chemicals, modulation of macromolecule synthesis and immune response, induction of phase II detoxifying enzymes, as well as its undesirable potential tumor-promoting activities. Understanding the molecular basis underlying these diverse biological actions of BHA is thus of great importance. Here we studied the pharmacokinetics, activation of signaling kinases and induction of phase II/III drug metabolizing enzymes/transporter gene expression by BHA in the mice. The peak plasma concentration of BHA achieved in our current study after oral administration of 200 mg/kg BHA was around $10\;{\mu}M$. This in vivo concentration might offer some insights for the many in vitro cell culture studies on signal transduction and induction of phase II genes using similar concentrations. The oral bioavailability (F) of BHA was about 43% in the mice. In the mouse liver, BHA induced the expression of phase II genes including NQO-1, HO-1, ${\gamma}-GCS$, GST-pi and UGT 1A6, as well as some of the phase III transporter genes, such as MRP1 and Slco1b2. In addition, BHA activated distinct mitogen-activated protein kinases (MAPKs), c-Jun N-terminal kinase (JNK), extracellular signal-regulated protein kinase (ERK), as well as p38, suggesting that the MAPK pathways may play an important role in early signaling events leading to the regulation of gene expression including phase II drug metabolizing and some phase III drug transporter genes. This is the first study to demonstrate the in vivo pharmacokinetics of BHA, the in vivo activation of MAPK signaling proteins, as well as the in vivo induction of Phase II/III drug metabolizing enzymes/transporters in the mouse livers.

• 한국미생물·생명공학회지
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• 제48권1호
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• pp.79-89
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• 2020

본 연구는 단풍잎돼지풀 70% 에탄올 추출물의 항염증 활성을 검증하기 위해 수행되었다. 단풍잎돼지풀 70% 에탄올 추출물의 전자공여능 측정과 ABTS⁺ 라디칼 소거능을 측정한 결과, 1,000 ㎍/ml 농도에서 각각 84.1%와 92.5%의 효과를 나타냈고, 수렴활성 측정을 한 결과 1,000 ㎍/ml 농도에서 94.7%의 효과를 보였다. 단풍잎돼지풀 70% 에탄올 추출물의 항염증 효과를 측정하기 위해 lipopolysaccharide (LPS)로 유도된 RAW 264.7세포를 사용하여 효과를 측정하였다. 세포에서 단풍잎돼지풀 추출물의 세포독성을 측정하기 위해 MTT assay를 수행하였다. 그 결과, 500 ㎍/ml 농도에서 90% 이상의 생존율을 보였다. Nitric oxide 생산을 억제하는 효과를 측정한 결과, 단풍잎돼지풀 추출물에서 농도가 증가할수록 NO 생성이 감소되는 효과를 확인하였다. 단풍잎돼지풀 추출물의 단백질 발현효과를 western blot을 통해 25, 50, 100 ㎍/ml 농도에서 측정하였고, 양성 대조군으로 β-actin을 사용하였다. 그 결과, inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2 단백질 발현효과는 100 ㎍/ml 농도에서 8.6%, 25.1%의 감소됨을 확인하였다. ERK1/2, p38, JNK와 Iκ-Bα의 단백질발현 효과는 인산화를 통해 확인하였고, 농도의존적으로 감소하였음을 확인하였다. mRNA 발현 억제 효과를 RT-PCR을 통해 25, 50, 100 ㎍/ml의 농도에서 측정하였고, 양성 대조군으로 GAPDH를 사용하였다. 그 결과, LPS로 유도된 대식세포에서 iNOS, COX-2, interleukin (IL)-1β, IL-6, TNF-α의 mRNA 발현 억제 효과는 농도가 증가할수록 발현이 감소됨을 확인하였다. 결론적으로 단풍잎돼지풀 추출물은 염증을 억제할 수 있는 가능성이 있는 항염증 소재의 가능성을 증명하였다.

• 한국식품영양과학회지
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• 제46권6호
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• pp.671-680
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• 2017

본 연구 결과 검은콩 물 에탄올 추출물과 Lactobacillus rhamnosus GG(LGG)와 Bifidobacterium animals subsp. lactis BB-12(BB-12)를 이용하여 발효시킨 발효검은콩 물 에탄올 추출물의 발효 전과 후의 성분변화를 분석하고, 검은콩과 검은콩 발효 추출물이 모유두 세포 성장에 미치는 효과를 알아보기 위해 primary 인간 모유두 세포(HFDPC)에 검은콩 추출물(BWE, BEE)과 발효검은콩 추출물(BWE-F, BEE-F)을 다양한 농도로 처리하여 세포독성을 확인하였으며, 이를 바탕으로 적정 처치 농도를 결정하여 모발 성장촉진(VEGF와 KGF/FGF7)과 억제($TGF-{\beta}1$과 AR)에 관여하는 유전자 발현을 mRNA 수준에서 확인하였다. 나아가 검은콩 추출물이 HFDPC의 생존 촉진에 미치는 영향을 Akt와 Erk의 인산화 활성을 통해 비교 분석하였다. LGG와 BB-12를 이용하여 발효시킨 검은콩은 발효가 진행됨에 따라 pH, 총폴리페놀, 총당 환원당의 함량이 감소하였으며, 검은콩 4종 추출물(BWE, BEE, BWE-F, BEE-F) 중 BWE, BEE, BWE-F가 VEGF의 mRNA 발현을 증가시키고, 모든 처리군에서 KGF/FGF7의 mRNA 발현을 유의적으로 증가시킴을 확인할 수 있었다. 나아가 BWE, BEE, BWE-F가 Erk의 활성을 증가시켰음을 확인할 수 있었다. 따라서 검은콩 물 추출물과 검은콩 에탄올 추출물, 그리고 발효검은콩 물 추출물이 인간 모유두 세포의 세포성장에 모발 성장 촉진인자의 활성과 Erk의 활성화 등을 통한 기전으로 긍정적인 영향을 미침으로써 모발 성장 및 모발 건강을 위한 기능성원료로서의 가능성을 확인할 수 있었으며, 유산균 2종(LGG, BB-12)을 이용한 발효검은콩이 검은콩과 비교하여 모발 성장 촉진 관련 단백질 발현에서는 유의적인 우월성을 가지지는 않음을 확인하였다. 추후 다른 유산균 균 총을 이용한 발효검은콩의 연구와 더불어 보다 정밀한 발효를 통한 검은콩 추출물의 성분과 조성의 변화를 바탕으로 한 모발 성장 및 모주기 관련 연구가 이루어져야 할 것이라 생각된다.

• The Korean Journal of Physiology and Pharmacology
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• 제17권3호
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• pp.203-208
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• 2013

As the abnormal proliferation of vascular smooth muscle cells (VSMCs) plays a critical role in the development of atherosclerosis and vascular restenosis, a candidate drug with antiproliferative properties is needed. We investigated the antiproliferative action and underlying mechanism of a newly synthesized naphthoquinone derivative, 5,8-dimethoxy-2-nonylamino-naphthalene-1,4-dione (2-nonylamino-DMNQ), using VSMCs treated with platelet-derived growth factor (PDGF). 2-Nonylamino-DMNQ inhibited proliferation and cell number of VSMCs induced by PDGF, but not epidermal growth factor (EGF), in a concentration-dependent manner without any cytotoxicity. This derivative suppressed PDGF-induced $[^3H]$-thymidine incorporation, cell cycle progression from $G_0/G_1$ to S phase, and the phosphorylation of phosphor-retinoblastoma protein (pRb) as well as the expression of cyclin E/D, cyclin-dependent kinase (CDK) 2/4, and proliferating cell nuclear antigen (PCNA). Importantly, 2-nonylamino-DMNQ inhibited the phosphorylation of PDGF receptor${\beta}$(PDGF-$R{\beta}$) enhanced by PDGF at $Tyr^{579}$, $Tyr^{716}$, $Tyr^{751}$, and $Tyr^{1021}$ residues. Subsequently, 2-nonylamino-DMNQ inhibited PDGF-induced phosphorylation of STAT3, ERK1/2, Akt, and $PLC{\gamma}1$. Therefore, our results indicate that 2-nonylamino-DMNQ inhibits PDGF-induced VSMC proliferation by blocking PDGF-$R{\beta}$ autophosphorylation, and subsequently PDGF-$R{\beta}$-mediated downstream signaling pathways.

• Journal of Microbiology and Biotechnology
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• 제30권11호
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• pp.1614-1625
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• 2020

A number of species of the genus Trichilia (Meliaceae) exhibit anti-inflammatory effects. However, the effect of Trichilia martiana C. DC. (TM) on lipopolysaccharide (LPS)-induced inflammation has not, to the best of our knowledge, yet been determined. Therefore, in the present study, the antiinflammatory effect of TM on LPS-stimulated RAW264.7 macrophages was evaluated. The ethanol extract of TM (TMEE) significantly inhibited LPS-induced nitric oxide (NO), prostaglandin 2 (PGE₂), inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). TMEE also reduced the levels of inflammatory cytokines, including tumor necrosis factor-alpha (TNF-α), interleukin (IL)-1β and IL-6. The upregulation of mitogen-activated protein kinases (MAPKs) and NF-κB activation was revealed to be downregulated following TMEE pretreatment. Furthermore, TMEE was indicated to lead to the nucleus translocation of nuclear factor erythroid-derived 2-related factor 2 (Nrf2) and the expression of heme oxygenase-1 (HO-1). In H292 airway epithelial cells, the pretreatment of TMEE significantly downregulated the production of LPS-stimulated IL-1β, and TMEE was indicated to increase the expression of HO-1. In animal models exhibiting LPS-induced acute lung injury (ALI), treatment with TMEE reduced the levels of macrophages influx and TNF-α production in the bronchoalveolar lavage fluid (BALF) of ALI mice. Additionally, TMEE significantly downregulated the activation of ERK, JNK and IκB, and upregulated the expression of HO-1 in the lungs of ALI mice. In conclusion, the results of the current study demonstrated that TMEE could exert a regulatory role in the prevention or treatment of the endotoxin-mediated inflammatory response.

• 생명과학회지
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• 제21권7호
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• pp.985-991
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• 2011

본 연구는 고지방식이로 인한 비만으로 불균형된 지질구성과 산화적 손상이 신경세포형성과 초기발현단백질에 미치는 생물학적 영향을 알아보고, 규칙적인 운동의 효과를 알아보기 위하여 실시하였다. 실험동물은 4주령 SD rat 수컷 30마리를 1주간의 적응기간을 둔 뒤, 15주간 고지방식이를 통해 비만으로 유도하였으며, high fat diet sedentary (HDS, n=15)와 high fat diet and training (HDT, n=15)으로 분류하여 연구하였다. 운동강도는 1~4주는 저강도, 5~8주는 중강도로 주5회 실시하였다. 8주 트레이닝 후 혈청지질, 8-OHdG, MDA, neurotrophic factor, 그리고 IEG를 분석하였다. 그 결과 TC와 TG에서 HDS와 HDT 사이 유의한 차이가 나타났다(p<0.05). 8-OHdG에서 HDT는 트레드밀 트레이닝 후에 HDS보다 낮게 나타났다(p<0.05). 해마에서 c-jun, BDNF 그리고 간에서 MDA의 단백질 발현은 HDT가 트레드밀 트레이닝 후 HDS보다 높게 나타났다(p<0.05). 결론적으로 8주간 트레드밀 훈련은 고지방식이 비만 유도 쥐의 혈청지질 성분의 불균형을 개선시키고, 조직과 혈청의 산화적 손상과 DNA 손상을 완화시켜 주어, 비만으로 인한 합병증 예방에 도움을 줄 수 있을 것으로 사료된다. 또한 NT의 발현을 증가시킴으로써 손상된 뇌기능과 신경세포의 생성 기전 활동에 긍정적 영향을 나타냄으로써 공간적 학습기능의 향상을 가져온 것으로 판단된다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 춘계학술발표회
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• pp.91-91
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• 2018

Poncirus Fructus (PF) is obtained by drying the trifoliate orange fruit belonging to the Rutaceae family. In our country of medicine, PF has been used as a treatment of indigestion, allergy and inflammation. But Mechanism and medical data for PF is insignificant. Recently, the effect of the study PF of biological activity was reported, such as anti- thrombosis, anti-bacteria, anti-virus, anti- allergic. We investigated that the effect of PF on anti-inflammatory in murine macrophage-like cell line Raw264.7 cells. Our results show that the expression level of Nitric Oxide (NO) and Matrix-metallopeptidase-9 (MMP-9) significantly decreased. Moreover, to determine the expression level of pro-inflammatory cytokines such as Tumor Necrosis Factor ($TNF-{\alpha}$) and Interleukin-6 (IL-6) and the phosphorylation pattern of signaling molecules of mitogen-activated protein kinase (MAPK) family, we performed ELISA and westren blot in Raw264.7 cells. In addition, nuclear factor-kappa B ($NF-{\kappa}B$) pathway was confirmed. PF extract inhibited the production of $TNF-{\alpha}$ and IL-6. The extract suppressed the phosphorylation of ERK1/2, JNK, and p38 MAPK, and the nuclear translocation of $NF-{\kappa}B$ p65 in activated cells. Our results suggest that PF can be used as a potential therapeutic agent or functional food to relieve inflammation.