• Title/Summary/Keyword: PD-1 receptor

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Differential Effect of Bovine Serum Albumin on Ginsenoside Metabolite-Induced Inhibition of ${\alpha}3{\beta}4$ Nicotinic Acetylcholine Receptor Expressed in Xenopus Oocytes

  • Lee, Jun-Ho;Jeong, Sang-Min;Lee, Byung-Hwan;Kim, Dong-Hyun;Kim, Jong-Hoon;Kim, Jai-Il;Lee, Sang-Mok;Nah, Seung-Yeol
    • Archives of Pharmacal Research
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    • v.26 no.10
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    • pp.868-873
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    • 2003
  • Ginsenosides, major active ingredients of Panax ginseng, that exhibit various pharmacological and physiological actions are transformed into compound K (CK) or M4 by intestinal microorganisms. CK is a metabolite derived from protopanaxadiol (PD) ginsenosides, whereas M4 is a metabolite derived from protopanaxatriol (PT) ginsenosides. Recent reports shows that ginsenosides might playa role as pro-drugs for these metabolites. In present study, we investigated the effect of bovine serum albumin (BSA), which is one of major binding proteins on various neurotransmitters, hormones, and other pharmacological agents, on ginsenoside $Rg_{2-}$, CK-, or M4-induced regulation of $\alpha3\beta4$ nicotinic acetylcholine (ACh) receptor channel activity expressed in Xenopus oocytes. In the absence of BSA, treatment of ACh elicited inward peak current ($I_{Ach}$) in oocytes expressing $\alpha3\beta4$ nicotinic ACh receptor. Co-treatment of ginsenoside $Rg_2$, CK, or M4 with ACh inhibited IAch in oocytes expressing $\alpha3\beta4$ nicotinic ACh receptor with reversible and dose-dependent manner. In the presence of 1% BSA, treatment of ACh still elicited $I_{Ach}$ in oocytes expressing $\alpha3\beta4$ nicotinic ACh receptor and co-treatment of ginsenoside $Rg_2$ or M4 but not CK with ACh inhibited $I_{Ach}$ in oocytes expressing $\alpha3\beta4$ nicotinic ACh receptor with reversible and dose-dependent manner. These results show that BSA interferes the action of CK rather than M4 on the inhibitory effect of $I_{Ach}$ in oocytes expressing $\alpha3\beta4$ nicotinic ACh receptor and further suggest that BSA exhibits a differential interaction on ginsenoside metabolites.

Role of ERK (Extracellular Signal Regulated Kinas) and PPARγ (Peroxisome Proliferator-Activated Receptor Gamma) on TGF-β1 Induced Human Endometrial Stromal Cell Decidualization (TGF-β1에 의하여 유도된 인간자궁내막의 탈락막화(Decidualization)에 있어서 ERK (Extracellular Signal Regulated Kinas)와 PPARγ (Peroxisome Proliferator-Activated Receptor Gamma)의 역할)

  • Chang, Hye Jin;Lee, Jae Hoon;Kim, Mi Ran;Hwang, Kyung Joo;Park, Dong Wook;Min, Churl K.
    • Clinical and Experimental Reproductive Medicine
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    • v.33 no.2
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    • pp.105-113
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    • 2006
  • Objective: To investigate the role of ERK and $PPAR{\gamma}$ on the $TGF-{\beta}1$ induced human endometrial stromal cell decidualization in vitro. Method: Endometrial stromal cells are cultured under the following condition: DMEM/F12 (10% FBS, 1 nM E2 and 100 nM P4). $TGF-{\beta}1$ (5 ng/ml), Rosiglitazone (50 nM), and PD98059 ($20{\mu}M$) were added according to experimental purposes. Trypan-Blue and hematocytometer were utilized to count cell number. Enzyme-linked immunosorbent assay (ELISA) and western blotting were utilized to detect proteins. Result: $TGF-{\beta}1$ inhibited proliferation of cultured human endometrial stromal cells and induced expression of PGE2 and prolactin. This effect was mediated by Smad and ERK activation. Administration of rosiglitazone, $PPAR{\gamma}$ agonist, prevented $TGF-{\beta}1$ effect on cell proliferation. Furthermore, Rosiglitazone inhibited $TGF-{\beta}1$ induced activation of ERK, consequently reduced PGE2 and prolactin production. Conclusion: $TGF-{\beta}1$ induced decidualization of endometrial stromal cell through Smad and ERK phosphorylation. $PPAR{\gamma}$ acts as a negative regulator of human ndometrial cell decidualization in vitro.

The role of p38 MAP kinase on RANKL regulation in mouse periodontal ligament fibroblasts (마우스 치주인대 섬유모세포에서 RANKL 조절에 대한 p38 MAP kinase의 역할)

  • Kim, Jae-Cheol;Cui, De-Zhe;Kim, Young-Joon
    • Journal of Periodontal and Implant Science
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    • v.37 no.sup2
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    • pp.311-323
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    • 2007
  • Receptor activation of nuclear factor ${\kappa}$ B ligand (RANKL)은 파골세포의 분화와 기능에 중요한 역할을 하는 단백질로 이들 물질의 조절에는 p38 MAP kinase가 관여한다. 그러나 치주인대 섬유모세포에서 RANKL 발현 시 p38 MAP kinase의 역할은 잘 알려져 있지 않다. 이에 이번 연구는 마우스 치주인대 섬유모세포의 $IL-1{\beta}-induced$ RANKL 발현과정에서 p38의 역할을 규명하고자 하여 다음과 같은 결과를 얻었다. 마우스 치주인대 섬유모세포에 $IL-1{\beta}$ (1ng/ml)의 자극은 수용성 RANKL의 합성을 증가시켰다. 수용성 RANKL의 합성은 p38 MAP kinase 억제제인 SB203580에 의해 농도 의존적으로 억제되었으나 다른 MAP kinase 억제제인 SP600125, JNK 억제제와 PD98059, ERK 억제제에 의해서는 수용성 RANKL의 합성이 조절되지 않았다. NF-kB 억제제에 의해서도 수용성 RANKL의 합성이 억제되지 않았다. RANKL 유전자의 발현은 $IL-1{\beta}$로 자극 시에는 대조군에 비해 약 5배의 발현 증가를 보였으나 SB203580으로 전처치 시 $IL-1{\beta}$ (1ng/ml)로 자극시보다 약 1.5배의 감소를 보였다. 그러나 SP600125, PD98059, 및 NF-kB 억제제로 전처치한 경우에는 $IL-1{\beta}$로 자극한 경우와 비슷한 수준을 보였다. $IL-1{\beta}$로 자극 시 RANKL 유전자의 반감기가 90분 이었으나 SB203580로 전처치 후 $IL-1{\beta}$로 자극 시 RANKL 유전자의 반감기는 60분으로 감소하였다. Cycloheximide 전처리 시 SB203580에 의한 RANKL 유전자 발현 억제가 관찰되지 않았다. 단백질 분석결과 p38 MAP kinase의 인산화 활성은 30분까지 증가하였으나 그 이후 감소하여 2시간째에는 그 발현이 미약하였다. SB203580로 전처치 후 $IL-1{\beta}$로 자극 시 p38 MAP kinase의 인산화 활성이 감소하였다. 이상의 결과는 p38 MAP kinase가 RANKL 유전자 조절에 중요한 역할을 담당하고 있음을 시사한다.

Muc5ac Gene Expression Induced by Cigarette Smoke is Mediated Via a Pathway Involving ERK1/2 and p38 MAPK (담배 연기에 의한 Muc5ac 유전자 발현에 관여하는 세포 내 신호 전달 경로로서의 ERK1/2와 p38 MAPK)

  • Kim, Yong Hyun;Yoon, Hyoung Kyu;Kim, Chi Hong;Ahn, Joong Hyun;Kwon, Soon Seog;Kim, Young Kyoon;Kim, Kwan Hyoung;Moon, Hwa Sik;Park, Sung Hak;Song, Jeong Sup;Cho, Kyung Sook
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.6
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    • pp.590-599
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    • 2005
  • Object : Cigarette smoking is a major cause of mucus hypersecretion, which is a pathophysiological feature of many inflammatory airway diseases. Mucins, which are an important part of the airway mucus, are synthesized from the Muc gene in airway epithelial cells. However, the signaling pathways for cigarette smoke-induced mucin synthesis are unknown. The aim of this study was to determine the signal pathway for smoking induced Muc5ac gene expression. Methods : A549 cells were cultured and transiently transfected with the Muc5ac promoter fragment. These cells were stimulated with 5% cigarette smoke extract (CSE) alone or with CSE after a pretreatment with various signal transduction pathway inhibitors (AG1478, PD98059 and SB203580). The Muc5ac promoter activity was examined using the luciferase reporter system, and the level of phosphorylated EGFR, ERK1/2, p38 MAPK and JNK were all examined using Western blot analysis. Muc5ac mRNA expression was also examined using reverse transcriptase polymerase chain reactions (RT-PCR). Results : 1. The peak level of luciferase activity of the Muc5ac promoter was observed at 5% concentration and after 3 hours of incubation with the CSE. The level of EGFR phosphorylation and the luciferase activity of the transfected cells caused by the CSE were significantly suppressed by AG1478 or PD98059 (P<0.01). 2. CSE phosphorylated ERK1/2 or p38 MAPK but not JNK. The Muc5ac mRNA expression level was increased by the CSE but that was suppressed by PD98059 or AG1478. 3. The CSE-induced phosphorylation of ERK1/2 was blocked by PD98059 and that of p38 MAPK was blocked by either PD98059 or SB203580. Either PD98059 or SB203580 suppressed the luciferase activity of the transfected cells (P<0.0001). Conclusion : The Muc5ac mRNA expression level was increased by the CSE. The increased CSE-induced transcriptional activity was mediated via EGF receptor activation, which led to ERK1/2 and p38 MAPK phosphorylation.

Aquaporin 8 Involvement in Human Cervical Cancer SiHa Migration via the EGFR-Erk1/2 Pathway

  • Shi, Yong-Hua;Tuokan, Talaf;Lin, Chen;Chang, Heng
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.15
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    • pp.6391-6395
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    • 2014
  • Overexpression of aquaporins (AQPs) has been reported in several human cancers. Epidermal growth factor receptor (EGFR)-extracellular signal-regulated kinases 1/2 (Erk1/2) are associated with tumorigenesis and cancer progression and may upregulate AQP expression. In this study, we demonstrated that EGF (epidermal growth factor) induces SiHa cells migration and AQP8 expression. Wound healing results showed that cell migration was increased by 2.79-1.50-fold at 24h and 48h after EGF treatment. AQP8 expression was significantly increased (3.33-fold) at 48h after EGF treatment in SiHa cells. An EGFR kinase inhibitor, PD153035, blocked EGF-induced AQP8 expression and cell migration and AQP8 expression was decreased from 1.59-fold (EGF-treated) to 0.43-fold (PD153035-treated) in SiHa. Furthermore, the MEK (MAPK (mitogen-activated protein kinase)/Erk (extracellular signal regulated kinase)/Erk inhibitor U0126 also inhibited EGF-induced AQP8 expression and cell migration. AQP8 expression was decreased from 1.21-fold (EGF-treated) to 0.43-fold (U0126-treated). Immunofluorescence microscopy further confirmed the results. Collectively, our findings show that EGF induces AQP8 expression and cell migration in human cervical cancer SiHa cells via the EGFR/Erk1/2 signal transduction pathway.

1,3-bisdicyanovinylindane 색소를 이용한 선택적 $Hg^{2+}$ 감지 특성

  • Kim, Su-Ho;Kim, Young-Sung;Kim, Sung-Hoon;Son, Young-A
    • Proceedings of the Korean Society of Dyers and Finishers Conference
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    • 2009.11a
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    • pp.19-20
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    • 2009
  • 최근 화학, 물리, 생명과학, 전기, 전자등의 다양한 분야에 활발하게 연구가 이루어지고 있는 초분자 화학은 선택적 분자인지를 위한 효율적인 골격구조와 나아가 다양한 계에 응용할 수 있다. 초분자 화학의 분자인지 과정의 특징은 일반적으로 수용체 (receptor 혹은 host)가 목표가 되는 기질 (substrate, analyte, 혹은 guest)에 대하여 선택적으로 식별하고 반응하는 것이다. 비공유 결합성 상호작용에 의하여 이루어지는 초분자 화학의 분자 인지 과정의 특징은 일반적으로 수용체 (receptor 혹은 host)가 목표가 되는 기질 (substrate, analyte, 혹은 guest)에 대하여 선택적으로 식별하고 반응하는 것이다. 이는 공유결합을 이용하는 분자화학과는 차별화 된 것이다. 수용체는 간단한 구조의 화합물 및 금속 이온들과 같은 기질과 가역적으로 상호 작용할 수 있는 착물을 형성한다. 최근들어 급격한 산업화가 진행되어 환경문제가 심각하게 대두 되어져 왔고, 그 중에서 특히 수은이나 카드뮴에 의한 질병, 납에 의한 중독 등 중금속에 의한 오염이 크게 나타남에도 불구하고, 현재 그러한 중금속을 검출함에 있어 많은 비용과 시간이 드는 문제점이 있다. 또한 우리에게 이로운 금속은 효율적 분석을 통해 환경계와 의료계에 많은 도움을 줄 것으로 사례되므로 화학센서 기술의 개발은 절실히 요구되어지고 있다. 이에 새로운 1,3-bisdicyanovinylindane 을 통해 $Hg^{2+}$ 금속의 감지 여부 알아보고, 그 특성을 파악하고자 한다. 1,3-indandion (2.18g, 14.9mmol), malononitrile (2.95g, 44.7mmol), ethanol 50ml를 20분간 상온에서 용해시킨다. 후에 sodium trihydrate acetate(3.05g)을 첨가한 후 5시간 동안 환류반응 시킨다. 이 과정에서 얻어진 용액을 필터과정을 통하여 에서 합성 반응 중에 생성된 불순물(1,3-dicyanovinylindane-1-one, monocondensation)을 제거한다. 필터과정을 통해 걸러진 미 반응 물질을 제�G 용반응욕액을 증류수(100ml)를 이용하여 희석시키고 난 후 염산을 이용, 산성화 시켜 고체 생성물을 얻어낸다. 이렇게 생성된 고체 생성물은 다시 필터 및 건조를 통하여 회색의 고체 화합물을 얻어낸다. 1,3-bisdicyanovinylindane과 금속이온에 대한 감응도를 확인하기 위하여 metanol/water(1:2)을 용매로 하여 금속이온의 농도를 변화시켜 발색특성을 살펴보았다. 본 색소화합물과 Hg2+에 대한 UV 흡광도 변화 적정결과와 그 화합물의 상태 살펴본 결과 금속이온이 0.2ml씩 더 참가되면서 색의 변화를 뚜렷하게 나타내었다. 반면 그 밖에 이온($Fe^{3+}$, $Ag^{2+}$, $Pd^{2+}$, $Zn^2$, $Fe^{2+}$, $Cu^{2+}$, $Pb^{2+}$)은 UV 흡광도 변화가 적거나 변화 자체가 없었다. 하지만 과량의 $Fe^{3+}$, $Ag^{2+}$, $Pd^{2+}$는 색상 변화를 나타내었으며,이와 같은 흡광도 변화는 금속에 따라 약간의 차이가 있지만, 420nm를 등흡수점으로 하여, 580nm의 파장 영역에 있는 흡수 밴드의 세기는 감소하는 반면 400nm 파장 영역에 있는 흡수 밴드의 세기가 증가하였다. 1,3-bisdicyanovinylindane 화합물은 다양한 생물계 및 환경계에서 요구되는 micro mol에서 milli mol 농도 영역의 $Hg^{2+}$ 이온의 선택적이고 민감한 검출과 정량적인 분석에 유용하게 사용될 수 있을 것이다.

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Effects of Anti-B7.1/B7.2 Antibodies on LPS-Stimulated Macrophages

  • Won, Tae-Joon;Huh, Yoon-Joo;Lim, Young-Tae;Song, Dong-Sup;Hwang, Kwang-Woo
    • Biomolecules & Therapeutics
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    • v.18 no.4
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    • pp.463-468
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    • 2010
  • T-cell activation depends on signals received by the T-cell receptor and CD28 co-stimulatory receptor. Since B7.1 and B7.2 molecules expressed on the surface of antigen presenting cells provide co-stimulatory signals through CD28 to T-cells, an inhibitor of CD28-B7.1/B7.2 binding has been proposed as a therapeutic agent for suppression of excessive T-cell activity. Although anti-B7.1/B7.2 antibodies are known to block B7.1 and B7.2 molecules, their effects on intracellular events in antigen presenting cells remain unclear. In this study, anti-B7.1/B7.2 antibodies decreased secretion of nitric oxide and pro-inflammatory cytokines such as TNF-$\alpha$, IL-$1{\beta}$, and IL-12 in LPS-activated RAW264.7 macrophage-like cells and peritoneal macrophages. Moreover, anti-B7.1/B7.2 antibodies inhibited $I{\kappa}B{\alpha}$ phosphorylation and down-regulated expression of co-stimulatory molecules including B7.1, B7.2, and PD-L1 in LPS-stimulated peritoneal macrophages. These findings suggest that CTLA4-Ig and anti-B7.1/B7.2 antibodies may be candidates to treat chronic inflammatory diseases and autoimmune responses caused by excessive activation of both T-cells and macrophages.

The Comparison of Commercial Serum-Free Media for Hanwoo Satellite Cell Proliferation and the Role of Fibroblast Growth Factor 2

  • In-sun Yu;Jungseok Choi;Mina K. Kim;Min Jung Kim
    • Food Science of Animal Resources
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    • v.43 no.6
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    • pp.1017-1030
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    • 2023
  • Fetal bovine serum (FBS), which contains various nutrients, comprises 20% of the growth medium for cell-cultivated meat. However, ethical, cost, and scientific issues, necesitates identification of alternatives. In this study, we investigated commercially manufactured serum-free media capable of culturing Hanwoo satellite cells (HWSCs) to identify constituent proliferation enhancing factors. Six different serum-free media were selected, and the HWSC proliferation rates in these serum-free media were compared with that of control medium supplemented with 20% FBS. Among the six media, cell proliferation rates were higher only in StemFlexTM Medium (SF) and Mesenchymal Stem Cell Growth Medium DXF (MS) than in the control medium. SF and MS contain high fibroblast growth factor 2 (FGF2) concentrations, and we found upregulated FGF2 protein expression in cells cultured in SF or MS. Activation of the fibroblast growth factor receptor 1 (FGFR1)-mediated signaling pathway and stimulation of muscle satellite cell proliferation-related factors were confirmed by the presence of related biomarkers (FGFR1, FRS2, Raf1, ERK, p38, Pax7, and MyoD) as indicated by quantitative polymerase chain reaction, western blotting, and immunocytochemistry. Moreover, PD173074, an FGFR1 inhibitor suppressed cell proliferation in SF and MS and downregulated related biomarkers (FGFR1, FRS2, Raf1, and ERK). The promotion of cell proliferation in SF and MS was therefore attributed to FGF2, which indicates that FGFR1 activation in muscle satellite cells may be a target for improving the efficiency of cell-cultivated meat production.

Oxytocin-induced endothelial nitric oxide dependent vasorelaxation and ERK1/2-mediated vasoconstriction in the rat aorta

  • Xu, Qian;Zhuo, Kunping;Zhang, Xiaotian;Zhang, Yaoxia;Xue, Jiaojiao;Zhou, Ming-Sheng
    • The Korean Journal of Physiology and Pharmacology
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    • v.26 no.4
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    • pp.255-262
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    • 2022
  • Oxytocin is a neuropeptide produced primarily in the hypothalamus and plays an important role in the regulation of mammalian birth and lactation. It has been shown that oxytocin has important cardiovascular protective effects. Here we investigated the effects of oxytocin on vascular reactivity and underlying the mechanisms in human umbilical vein endothelial cells (HUVECs) in vitro and in rat aorta ex vivo. Oxytocin increased phospho-eNOS (Ser 1177) and phospho-Akt (Ser 473) expression in HUVECs in vitro and the aorta of rat ex vivo. Wortmannin, a specific inhibitor of phosphatidylinositol 3-kinase (PI3K), inhibited oxytocin-induced Akt and eNOS phosphorylation. In the rat aortic rings, oxytocin induced a biphasic vascular reactivity: oxytocin at low dose (10-9-10-8 M) initiated a vasorelaxation followed by a vasoconstriction at high dose (10-7 M). L-NAME (a nitric oxide synthase inhibitor), endothelium removal or wortmannin abolished oxytocin-induced vasorelaxation, and slightly enhanced oxytocin-induced vasoconstriction. Atosiban, an oxytocin/vasopressin 1a receptor inhibitor, totally blocked oxytocin-induced relaxation and vasoconstriction. PD98059 (ERK1/2 inhibitor) partially inhibited oxytocin-induced vasoconstriction. Oxytocin also increased aortic phospho-ERK1/2 expression, which was reduced by either atosiban or PD98059, suggesting that oxytocin-induced vasoconstriction was partially mediated by oxytocin/V1aR activation of ERK1/2. The present study demonstrates that oxytocin can activate different signaling pathways to cause vasorelaxation or vasoconstriction. Oxytocin stimulation of PI3K/eNOS-derived nitric oxide may participate in maintenance of cardiovascular homeostasis, and different vascular reactivities to low or high dose of oxytocin suggest that oxytocin may have different regulatory effects on vascular tone under physiological or pathophysiological conditions.

Peroxisome proliferator-activated receptor γ is essential for secretion of ANP induced by prostaglandin D2 in the beating rat atrium

  • Zhang, Ying;Li, Xiang;Liu, Li-Ping;Hong, Lan;Liu, Xia;Zhang, Bo;Wu, Cheng-Zhe;Cui, Xun
    • The Korean Journal of Physiology and Pharmacology
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    • v.21 no.3
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    • pp.293-300
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    • 2017
  • Prostaglandin $D_2$ ($PGD_2$) may act against myocardial ischemia-reperfusion (I/R) injury and play an anti-inflammatory role in the heart. Although the effect of $PGD_2$ in regulation of ANP secretion of the atrium was reported, the mechanisms involved are not clearly identified. The aim of the present study was to investigate whether $PGD_2$ can regulate ANP secretion in the isolated perfused beating rat atrium, and its underlying mechanisms. $PGD_2$ (0.1 to $10{\mu}M$) significantly increased atrial ANP secretion concomitantly with positive inotropy in a dose-dependent manner. Effects of $PGD_2$ on atrial ANP secretion and mechanical dynamics were abolished by AH-6809 ($1.0{\mu}M$) and AL-8810 ($1.0{\mu}M$), $PGD_2$ and prostaglandin $F2{\alpha}$ ($PGF2{\alpha}$) receptor antagonists, respectively. Moreover, $PGD_2$ clearly upregulated atrial peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) and the $PGD_2$ metabolite 15-deoxy-${\Delta}12$, 14-$PGJ_2$ (15d-$PGJ_2$, $0.1{\mu}M$) dramatically increased atrial ANP secretion. Increased ANP secretions induced by $PGD_2$ and 15d-$PGJ_2$ were completely blocked by the $PPAR{\gamma}$ antagonist GW9662 ($0.1{\mu}M$). PD98059 ($10.0{\mu}M$) and LY294002 ($1.0{\mu}M$), antagonists of mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) and phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt) signaling, respectively, significantly attenuated the increase of atrial ANP secretion by $PGD_2$. These results indicated that $PGD_2$ stimulated atrial ANP secretion and promoted positive inotropy by activating $PPAR{\gamma}$ in beating rat atria. MAPK/ERK and PI3K/Akt signaling pathways were each partially involved in regulating $PGD_2$-induced atrial ANP secretion.