5-Lipoxygenase in monocytes emerges as a therapeutic target for intimal hyperplasia in a murine wire-injured femoral artery

Baek, S.E.,Jang, M.A.,Lee, S.J.,Park, S.Y.,Bae, S.S.,Kim, C.D. Elsevier/North Holland [etc.] 2017 Biochimica et biophysica acta Vol.1863 No.9

Given the importance of leukotrienes in vascular inflammation induced by local tissue injury, this study investigated the role for 5-lipoxygenase (5-LO) in monocytes in the development of intimal hyperplasia. As a mechanistic study, the importance of monocyte 5-LO in monocyte-macrophage differentiation with subsequent infiltration in neointima was evaluated. In a mouse model of wire-injured femoral artery, intimal hyperplasia started as early as 2wks after injury, and luminal area and blood flow were reduced due to increased neointima formation. Time-dependent increases in macrophage infiltration were observed in neointima and showed a positive relationship with neointima volume. In 5-LO-deficient (KO) mice or wild-type (WT) mice treated with an inhibitor of 5-LO activating protein (MK886, 1 and 10mg/kg), intimal hyperplasia and macrophage infiltration into neointima were reduced, but monocyte adhesion to injured luminal surface was not inhibited, which suggested 5-LO participates in monocyte-macrophage differentiation. In an in vitro study, monocyte-macrophage differentiation was found to be increased by high mobility group box 1 protein (HMGB1), but this effect was attenuated in cells isolated from 5-LO-KO mice. Furthermore, macrophage infiltration and intimal hyperplasia were more prominent in 5-LO-KO mice transplanted with monocytes from WT mice than in 5-LO-KO mice transplanted with monocytes from 5-LO-KO mice. Taken together, it was suggested that 5-LO in monocytes played a pivotal role in monocyte-macrophage differentiation and subsequent infiltration of macrophage in neointima, leading to vascular remodeling after vascular injury.

단핵구의 부착, 분화 및 SR-A 발현에 대한 철 결핍의 영향

서검석(Geom Seog Seo),최석채(Suck Chei Choi),최은영(Eun Young Choi),김태현(Tae Hyeon Kim),나용호(Yong Ho Nah),오재민(Jae Min Oh),김상욱(Sang Wook Kim),전창덕(Chang-Duk Jun) 대한해부학회 2004 Anatomy & Cell Biology Vol.37 No.6

철 항산성의 유지는 세포의 증식 및 분화에 필수적이며, 면역 기능 조절에도 중요한 역할을 한다. 단핵구와 큰포식세포는 급만성 염증에 있어 숙주 방어의 중요 역할을 한다. 또한 단핵구 및 큰포식세포는 세망내피계통의 다른 세포들과 더불어 철의 저장에 있어 중요한 역할을 한다. 이러한 단핵구인 THP-1 세포에서 철 결핍이 발생될 때 형태학적 및 기능적 변화에 대한 연구가 없어 알아보았다. Desferrioxamine (DFO)을 처리한 단핵구에서는 세포 부착능이 증가하였고 미분간섭 현미경(Differential interference contrast, DIC)에서 DFO를 처리한 세포의 거짓발 연장(pseudopodial extension)이 관찰되었다. Giemsa 염색에서 핵세포질 비율(N/C ratio)의 변화를 보였다. 또한 fluorescence-activated cell sorter (FACS)와 rhodamine-phalloidin 염색에서 DFO에 의한 단핵구의 탐식능 증가가 관찰되였다. 세포 분화 인자인 scavenger receptor-A (SR-A)의 발현이 DFO 처리한 단핵구에서 증가되었다. 이러한 결과는 DFO에 의한 철 결핍 상태가 단핵구의 큰포식세포양 세포로 분화를 촉진시키며, 이는 염증에서 숙주 면역 반응을 약화시킬 것으로 추측된다. Maintenance of cellular iron homeostasis is a prerequisite for proliferation and differentiation of cells, and is also a central role in the regulation of immune function. Monocyte-macrophages play an important roles in host defense, particularly in the inflammatory process of acute and chronic disease. The reason that an iron is important in these cell is because an iron is indispensable in a generation of hydroxyl radical for bacterium killing. Because of the role of iron in the monocytic THP-1 cell differentiation is not become clear, we investigated whether THP-1 cell can differentiate to macrophage-like cell using of iron and iron chelator which cause iron depletion. The cell differentiation was not able to observe by iron treatment, by the way, the cell adhesion was increased in DFO treated monocyte and cellular pseodopodial extension, change of a nucleus-cytoplasmic ratio were showed in Differential interference contrast (DIC) and Giemsa staining, and it was inhibited by ferric citrate (FC). Increased polystyrene bead phagocytosis by DFO treatment of THP-1 cell were detected through FACS and rhodamine-phallodin staining. The SR-A expression, which was a cell differentiation marker, was increased by DFO treatment of THP-1 cell. These results suggest that iron depletion by DFO can promote THP-1 cell diffentiation into macrophage-like cell, and this may carrying out important role in the immune response.

Red ginseng extract regulates differentiation of monocytes to macrophage and inflammatory signalings in human monocytes

강보빈,김채영,황지수,선소정,양현원,서형주,최현선 한국식품과학회 2019 Food Science and Biotechnology Vol.28 No.6

This study was aimed to investigate the effect ofred ginseng extract (RGE) on monocyte to macrophagedifferentiation and inflammatory signalings in THP-1human monocytes. In HPLC analysis, RGE containedsaponin level of 516 lg/mg (extract) with 14 ginsenosides. RGE effectively suppressed the monocyte-to-macrophagedifferentiation induced by phorbol 12-myristate 13-acetated(PMA) by inhibiting the THP-1 cell adhesion. Thisresult is evidenced by the down-regulation of cluster ofdifferentiation molecule b (CD11b) and CD36. RGE significantlyreduced translocation of nuclear factor kappalight-chain-enhancer of activated B cells (NF-jB) (78%),while cytosolic NF-jB was increased (53%), comparedwith LPS group. In addition, RGE significantly increasedthe protein abundance of nuclear factor (erythroid-derived2)-like 2 (Nrf2) and its target protein, hemoxygenase-1(HO-1), but, Kelch-like ECH-associated protein 1(KEAP1), a negative regulator of Nrf2, was greatlydecreased by RGE. Furthermore, RGE effectivelymediated the regulation of Nrf2 level in nucleus andcytoplasm of THP-1.

The orphan nuclear receptor SHP inhibits apoptosis during the monocytic differentiation by inducing p21WAF1

KyeongJin Kim,Yoon Ha Choi,김형회,정재훈 생화학분자생물학회 2009 Experimental and molecular medicine Vol.41 No.6

Small heterodimer partner (SHP) is an atypical member of nuclear receptor superfamily that lacks a DNA-binding domain. In previous study, we showed that SHP, c-jun, p65 of NF-κB subunits, and p21WAF1 expression was increased during monocytic differentiaton with the exposure of human leukemia cells to a differentiation agent, PMA. In this study, c-Jun and p65 were shown to mediate the transcriptional activation of the SHP promoter. In addition, SHP induced the cell cycle regulatory protein levels and cooperatively increased an induction of p21WAF1 expression with p65. Furthermore, SHP protected differentiated cells from etoposide-induced cellular apoptosis through the induction and cytoplasmic sequestration of p21WAF1. Complex formation between SHP and p21WAF1 was demonstrated by means of coimmunoprecipitation. These results suggest that SHP prolongs a cellular survival of differentiating monocytes through the transcriptional regulation of target genes of cell survival and differentiation.

Apoptosis: role in myeloid cell development

Shilpa Sarvothaman,Ram Babu Undi,Satya Ratan Pasupuleti,Usha Gutti,Ravi Kumar Gutti 대한혈액학회 2015 Blood Research Vol.50 No.2

Hematopoiesis is the process that generates blood cells in an organism from the pluripotent stem cells. Hematopoietic stem cells are characterized by their ability to undergo self-renewal and differentiation. The self-renewing ability ensures that these pluripotent cells are not depleted from the bone marrow niche. A proper balance between cell death and cell survival is necessary to maintain a homeostatic condition, hence, apoptosis, or programmed cell death, is an essential step in hematopoiesis. Recent studies, however, have introduced a new aspect to this process, citing the significance of the apoptosis mediator, caspase, in cell development and differentiation. Extensive research has been carried out to study the possible role of caspases and other apoptosis related factors in the developmental processes. This review focuses on the various apoptotic factors involved in the development and differentiation of myeloid lineage cells: erythrocytes, megakaryocytes, and macrophages.

Apoptosis: role in myeloid cell development

Shilpa Sarvothaman,Ram Babu Undi,Satya Ratan Pasupuleti,Usha Gutti,Ravi Kumar Gutti 대한혈액학회 2015 Blood Research Vol.50 No.2

Hematopoiesis is the process that generates blood cells in an organism from the pluripotent stem cells. Hematopoietic stem cells are characterized by their ability to undergo self-renewal and differentiation. The self-renewing ability ensures that these pluripotent cells are not depleted from the bone marrow niche. A proper balance between cell death and cell survival is necessary to maintain a homeostatic condition, hence, apoptosis, or programmed cell death, is an essential step in hematopoiesis. Recent studies, however, have introduced a new aspect to this process, citing the significance of the apoptosis mediator, caspase, in cell development and differentiation. Extensive research has been carried out to study the possible role of caspases and other apoptosis related factors in the developmental processes. This review focuses on the various apoptotic factors involved in the development and differentiation of myeloid lineage cells: erythrocytes, megakaryocytes, and macrophages.

Galectin-4 Interaction with CD14 Triggers the Differentiation of Monocytes into Macrophage-like Cells via the MAPK Signaling Pathway

홍소희,신준섭,정현우,박정규 대한면역학회 2019 Immune Network Vol.19 No.3

Galectin-4 (Gal-4) is a β-galactoside-binding protein mostly expressed in the gastrointestinal tract of animals. Although intensive functional studies have been done for other galectin isoforms, the immunoregulatory function of Gal-4 still remains ambiguous. Here, we demonstrated that Gal-4 could bind to CD14 on monocytes and induce their differentiation into macrophage-like cells through the MAPK signaling pathway. Gal-4 induced the phenotypic changes on monocytes by altering the expression of various surface molecules, and induced functional changes such as increased cytokine production and matrix metalloproteinase expression and reduced phagocytic capacity. Concomitant with these changes, Gal-4-treated monocytes became adherent and showed elongated morphology with higher expression of macrophage markers. Notably, we found that Gal-4 interacted with CD14 and activated the MAPK signaling cascade. Therefore, these findings suggest that Gal-4 may exert the immunoregulatory functions through the activation and differentiation of monocytes.

Arabinogalactan-type Polysaccharides (APS) from Cordyceps militaris Grown on Germinated Soybeans (GSC) Induces Innate Immune Activity of THP-1 Monocytes through Promoting Their Macrophage Differentiation and Macrophage Activity

박동기,Toshimitsu Hayashi,박혜진 한국식품과학회 2012 Food Science and Biotechnology Vol.21 No.5

In this study, the immunemodulatory activities of arabinogalactans-type polysaccharide (APS) from Cordyceps militaris grown on germinated soybeans was evaluated in THP-1 human monocytes. The cellular size and the number of intracellular organelles of THP-1monocytes were increased by APS. Also, APS-treated THP-1 cells significantly developed cellular adherence and macrophagic differentiation to the culture plate surface. APS noticeably enhanced phagocytic activity of THP-1cells against IgG-FITC-latex beads. APS induced the level of TNF-α, IL-12 p40, and IL-8 as well as TLR2 and TLR4mRNAs. APS can be developed as a promising immunmodulating agent with macrophage-activating properties.

홍삼 비사포닌 분획의 단핵세포 분화와 염증반응에 대한 억제효과

강보빈(Bobin Kang),김채영(Chae Young Kim),황지수(Jisu Hwang),최현선(Hyeon-Son Choi) 한국식품과학회 2019 한국식품과학회지 Vol.51 No.1

본 연구에서는 홍삼 비사포닌 분획(NSF)의 항 염증 효과를 마우스 대식세포와 인간유래 단핵세포에서 확인하였다. NSF는 마우스 대식세포에서 LPS로 유도된 NO, iNOS 그리고 COX-2의 양 뿐만 아니라 IL-6, TNF-α, MCP-1과 같은 염증성 싸이토카인의 생성량을 유의적으로 감소시켰다. 인간 유래 단핵세포에서는 PMA에 의해 유도되는 대식세포로의 분화를 효과적으로 억제하면서 분화인자인 CD11β와 CD36의 발현을 유의적으로 감소시켰다. 마우스 대식세포에서와 마찬가지로 염증성 싸이토카인들의 생성량 또한 감소하였는데, 이러한 NSF의 항 염증 효과는 두 전사인자의 조절작용에 의한 것으로 사료된다. 즉 NSF는 NF-κB의 핵으로 이동을 감소시킴으로써 전사활성을 억제하여 염증성 싸이토카인들의 발현을 저해하고 이와 반대로 Nrf2의 발현과 핵으로의 이동을 증가시켜 항산화 효소이면서 항 염증 작용을 나타내는 HO-1의 발현을 촉진하는 것으로 관찰되었다. 따라서 NSF는 NF-κB와 Nrf2의 두 가지 신호전달체계를 조절함으로써 항 염증 작용을 나타냈으며 이를 홍삼 NSF의 항 염증 기작으로 보고하는 바이다. The aim of this study was to investigate the effects of red ginseng-derived non-saponin fraction (NSF) on inflammatory responses and monocyte-to-macrophage differentiation in RAW264.7 and THP-1. NSF effectively inhibited inflammatory responses by downregulating nitric oxide (NO) production and protein levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). NSF (2000 μg/mL) decreased the levels of NO, iNOS, and COX-2 by 33, 83, and 64%, respectively. NSF inhibited the differentiation of monocyte-to-macrophage by decreasing cell adherence along with downregulation of the cluster of differentiation molecule 11β (CD11β) and CD36. In addition, pro-inflammatory cytokines, such as tumor necrosis factor-alpha, interleukin 6, and monocyte chemoattractant protein 1 (MCP-1), were significantly reduced with NSF treatment. The NSF-mediated inhibition of inflammatory responses was due to the regulation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and nuclear factor (erythroid-derived 2)-like 2 (Nrf2). NSF effectively suppressed the translocation of NF-κB into the nucleus, while nuclear Nrf2 and its target protein, heme oxygenase-1, levels were significantly increased.

7-Oxygenated cholesterol molecules differentially affect the expression of zonula occludens-1 in vascular smooth muscle cells and monocyte/macrophage cells

Cho, Hyok-rae,Kim, Koanhoi,An, Won G.,Eo, Seong-Kug,Bae, Sun Sik,Kim, Chi Dae,Son, Yonghae Elsevier 2018 Biochemical and biophysical research communication Vol.497 No.2

<P><B>Abstract</B></P> <P>To investigate the effects of 7-oxygenated cholesterol molecules on the expression of tight junction proteins, we examined the outcomes effects of 7-ketocholesterol (7K), 7α-hydroxycholesterol (7αOHChol) and 7β-hydroxycholesterol (7βOHChol) on the expression of the tight-junction protein zonula occludens-1 (ZO-1) using vascular cells. Vascular smooth muscle cells (VSMCs) constitutively express ZO-1, and this expression remained unaffected in the presence of cholesterol. However, the level of ZO-1 protein decreased after exposure to 7K and, to a lesser extent, 7αOHChol and 7βOHChol. ZO-1 was translocated to the nucleus following treatment with 7K; this translocation was inhibited by z-VAD-fmk, a pan-caspase inhibitor. ZO-1 protein was found to disintegrate in the aorta of ApoE knockout mice fed a high cholesterol diet, whereas it remained intact in the wild-type control. THP-1 monocyte/macrophage cells, which show no expression of ZO-1, were not influenced by treatment with cholesterol, 7K, and 7βOHChol. However, the treatment of THP-1 cells with 7αOHChol resulted in ZO-1 expression, which largely remained localized on the cytoplasmic membrane. These results indicate the varying effects of 7-oxygenated cholesterol molecules on the expression and localization of ZO-1 depending on cell types, and suggest the contribution of 7-oxygeneted cholesterol molecules to the structural alteration of tight junctions.</P>