탄저 치사독소 처리에 의한 생쥐 대식세포의 단백질체 발현 양상 분석

정경화,서귀문,김성주,김지천,오선미,오광근,채영규,Jung Kyoung-Hwa,Seo Giw-Moon,Kim Sung-Joo,Kim Ji-Chon,Oh Seon-Mi,Oh Kwang-Geun,Chai Young-Gyu 한국미생물학회 2005 미생물학회지 Vol.41 No.4

탄저 치사독소는 생쥐 대식세포 (RAW 264.7)의 유전자 발현에 많은 변화를 초래한다. 이들 변화를 초래하는 치사독소의 역할은 아직 확실하게 밝혀지지 ???았다. 본 연구에서는, 치사독소가 처리된 생쥐 대식세포의 단백질 프로파일을 이차원 전기영동으로 분석하였고, MALDI-TOF 질량분석기를 사용하여 해당 단백질의 질량을 측정하였다. 펩타이드 질량 분석 데이터는 ProFound 데이터베이스를 이용하여 동정하였다. 차별화되어 발현된 단백질 중에서 절단된 mitogen-activated protein kinase kinase (Mek1)와 glucose-6-phosphate dehydrogenase (G6PD)가 치사독소 처리된 대식세포에서 각각 증가하였다. 치사독소를 처리하였을 경우, Mek1의 절단은 신호전달과정을 방해하고, 증가된 G6PD는 생성된 활성산소로부터 세포를 보호하는 역할을 하는 것으로 보인다. 단백질체 분석기술은 치사독소처리에 의한 생쥐 대식세포의 세포사멸 관련 단백질을 동정하는데 도움을 주어, 치사독소의 잠정적인 기질을 찾는데 유용할 것이다. Intoxication of murine macrophages (RAW 264.7) with the anthrax lethal toxin (LeTx 100 ng/ml) results in profound alterations in the host cell gene expression. The role of LeTx in mediating these effects is unknown, largely due to the difficulty in identifying and assigning function to individual proteins. In this study, we have used two-dimensional polyacrylamide gel electrophoresis to analyze the protein profile of murine macrophages treated with the LeTx, and have coupled this to protein identification using MALDI-TOF mass spectrometry. Interpretation of the peptide mass fingerprint data has relied primarily on the ProFound database. Among the differentially expressed spots, cleaved mitogen-activated protein kinase kinase (Mek1) and glucose-6-phosphate dehydrogenase were increased in the LeTx treated macrophages. Mek1 acts as a negative element in the signal transduction pathway, and G6PD plays the role for the protection of the cells from the hyper-production of active oxygen. Our results suggest that this proteomic approach is a useful tool to study protein expression in intoxicated macrophages and will contribute to the identification of a putative substrate for LeTx.

八物湯이 생쥐 임신중기에 胸線細布 및 腹腔 Macrophage로부터 cytokines 생성에 미치는 영향

유동렬 대한동의병리학회 2001 동의생리병리학회지 Vol.15 No.3

The purpose of this research was to investigate effects of Palmul-tang water extract(PMTE) on cytokines production from murine thymocytes and peritoneal macrophages during the middle stage of pregnancy. PMTE (500mg/kg) was administered p.o. once a day for 7 days, and then thymocytes and peritoneal macrophages were separated. At the middle stage of pregnant mice, the production of interferon-γ, interleukin-2 and interleukin-4 from thymocytes was enhanced, but was decreased by administration of PMTE. The production of tumor necrosis factor-α and interleukin-1β from peritoneal macrophages was decreased, but tumor necrosis factor-α was enhanced and interleukin-1β was decreased by PMTE. The production of nitric oxide from peritoneal macrophages was enhanced, but was decreased by PMTE. The proliferation of thymocytes was decreased by PMTE. These results suggest that PMTE has the regulative action on immune function of thymocytes and peritoneal macrophages at the middle stage of pregnant mice.

Oleamide suppresses inflammatory responses in LPS-induced RAW264.7 murine macrophages and alleviates paw edema in a carrageenan-induced inflammatory rat model

Moon, Sung-Min,Lee, Seul Ah.,Hong, Joon Ho,Kim, Jae-Sung,Kim, Do Kyung,Kim, Chun Sung Elsevier 2018 INTERNATIONAL IMMUNOPHARMACOLOGY Vol.56 No.-

<P><B>Abstract</B></P> <P>Oleamide compounds purified from green algae have been used for the prevention and treatment of atherosclerosis, thrombosis, arthritis, and cancer. They function through their metabolic conversion into prostaglandins, thromboxanes, and leukotrienes. However, the actual mechanism of action has not been well characterized. To investigate the underlying anti-inflammatory activity and associated mechanisms, oleamide purified from <I>Codium fragile</I> was studied using RAW264.7 murine macrophages and a carrageenan-induced inflammatory rat model. Our results indicate that pre-treatment of RAW264.7 cells with oleamide significantly suppressed LPS-induced nitrite production and PGE<SUB>2</SUB> secretion. Oleamide inhibited LPS-induced iNOS and COX-2 mRNA and protein expression. It also inhibited the LPS-induced production of inflammatory cytokines such as tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-6. In addition, oleamide prevented the nuclear translocation of NF-κB by suppressing the phosphorylation of the inhibitor of kappa B (IκB)-α. Oleamide also suppressed the phosphorylation of mitogen-activated protein kinases such as ERK1/2 and JNK. Furthermore, inhibition of paw swelling (%) was suppressed 2 h after the intraperitoneal injection of oleamide (20 mg/kg, body weight) in a carrageen-induced rat model. Therefore, our results suggest that oleamide can be used as a single ingredient treatment for inflammatory diseases.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Oleamide significantly suppressed LPS-induced nitrite production, and PGE<SUB>2</SUB> secretion in RAW264.7 murine macrophages. </LI> <LI> Oleamide inhibited LPS-induced pro-inflammatory cytokines, including TNF-α, IL-6, and IL-1β in RAW264.7 murine macrophages. </LI> <LI> Oleamide inhibits the translocation of NF-κB p65 to the nucleus by inhibiting IκB-α phosphorylation. </LI> <LI> Oleamide effectively inhibits ERK1/2 and JNK phosphorylation during LPS-induced inflammatory response. </LI> <LI> Oleamide suppressed the carrageenan-induced paw edema thickness at 2 h after injection of carrageenan. </LI> </UL> </P>

Inhibition of Lipopolysaccharide-induced Inducible Nitric Oxide (iNOS) mRNA Expression and Nitric Oxide Production by Higenamine in Murine Peritoneal Macrophages

Lee, Hoi Young,Lee, Jang-Soon,Kim, Eun Ju,Han, Jeung Whan,Lee, Hyang Woo,Kang, Young Jin,Chang, Ki Churl 성균관대학교 약학연구소 1999 成均藥硏論文集 Vol.11 No.-

Nitric oxide synthesized by inducible nitric oxide synthase (iNOS) has been implicated as a mediator of inflammation in rheumatic and autoimmune diseases. The effects of higenamine, a tetrahydroisoquinoline compound, on induction of NOS by bacterial lipopolysaccharide(LPS) were examined in murine peritoneal macrophages. LPS-induced nitrite/nitrate production was markedly inhibited by higenamine which at 0.01 mM, decreased nitrite/nitrate levels by 48.7±4.4%. This was comparable to the inhibition of LPS-induced nitrite/nitrate production by tetrandrin (49.51±2.02%) at the same concentration. Northern and Western blot analysis of iNOS expression demonstrated that iNOS expression was significantly attenuated following co-incubation of peritoneal macrophages with LPS (10 ㎍/ml; 18hrs) and higenamine (0.001, 0.01 mM; 18hrs). These results suggest that higenamine can inhibit LPS-induced expression of iNOS mRNA in murine peritoneal macrophages. The clinical implications of these findings remain to be established.

한국산 겨우살이 열매 추출물의 마우스 복강 대식세포 면역활성화 효과

이정림,전영하,양효선,이경복,송경식,강태봉,김종배,유영춘 한국생약학회 2010 생약학회지 Vol.41 No.2

Mistletoe (Viscum album) is a common name for many species of semi-parasitic plants which grow on deciduous trees all over the world. In this study, the immunomodulatory activity of the water-extract of Korean mistletoe fruits (KMF-WE), was investigated on murine peritoneal macrophages. The culture supernatants of KMF-WE-stimulated murine peritoneal macrophages showed the increased production of IFN-g, IL-1b and TNF-a, in a dose-dependent manner. KMF-WE also induced chemokine production from murine peritoneal macrophages such as RANTES, MCP-1, MIP-1a and MIP-1b, as well as nitric oxide (NO) production, in a dose-dependent manner. The gel filtration fraction revealed F-1, which is the early-eluted and high molecular weight product, is the major fraction of KMF-WE to activate the murine peritoneal macrophage to induce cytokines, chemokines and NO. The nature of F-1 fraction needs to be examined in detail in further studies to define the regulatory mechanisms of cytokine or chemokine induction by KMF-WE on macrophages. These results suggest that KMF-WE possess a potent immunostimulant activity and can be a promising candidate available for development of immunomodulators.

Inhibition of Lipopolysaccaride-induced Inducible Nitric Oxide (iNOS) mRNA Expression and Nitric Oxide Production by Higenamine in Murine Peritoneal Macrophages

Lee, Hoi-Young,Lee, Jang-Soon,Kim, Eun-Ju,Han, Jeung-Whan,Lee, Hyang-Woo,Kang, Young-Jin,Chang, Ki-Churl The Pharmaceutical Society of Korea 1999 Archives of Pharmacal Research Vol.22 No.1

Nitric oxide synthesized by inducible nitric oxide synthase (iNOS) has been implicated as a mediator of inflammation in rheumatic and autoimmune diseases. The effects of higenamine, a tetrahydroisoquinoline compound, on induction of NOS by bacterial lipopolysaccaride (LPS) were examined in murine peritoneal macrophages. LPS-induced nitrite/nitrate production was markedly inhibited by higenamine which at 0.01 mM, decreased nitrite/nitrate levels by $48.7{\pm}4.4%$This was comparable to the inhibition of LPS-induced nitrite/nitrate production by tetrandrin ($49.51{\pm}2.02%$). at the same concentration. Northern and Western blot analysis of iNOS expression demonstrated that iNOS expression was significantly attenuated following co-incubation of peritoneal macrophages with LPS (10 $\mu\textrm{g}$/m;; 18hrs) and higenamine (0.001, 0.,01 mM; 18hrs). These results suggest that higenamine can inhibit LPS-induced expression of iNOS mRNA in murine peritoneal macrophages. The clinical implications of these findings remain to be established.

Pulchellamin G, an amino acid-sesquiterpene lactone, from Saussurea pulchella suppresses lipopolysaccharide-induced inflammatory responses via heme oxygenase-1 expression in murine peritoneal macrophages

Lee, D.S.,Choi, H.G.,Wan Woo, K.,Kang, D.G.,Lee, H.S.,Oh, H.,Ro Lee, K.,Kim, Y.C. North-Holland ; Elsevier Science Ltd 2013 european journal of pharmacology Vol.715 No.1

Saussurea pulchella (Asteraceae) is widely distributed in Korea and has been used in Korean folk medicine for the treatment of inflammation, hypertension, hepatitis, and arthritis. Pulchellamin G is an amino acid-sesquiterpene lactone conjugate isolated from S. pulchella. In the present study, we focused on the anti-inflammatory effect of pulchellamin G, which acts by inducing the expression of heme oxygenase (HO)-1. HO-1 plays important roles in cytoprotection since it has antioxidant, anti-inflammatory, antiproliferative, and antiapoptotic properties. Pulchellamin G inhibited the mRNA and protein expression of inducible nitric oxide synthase (iNOS), iNOS-derived nitric oxide (NO), and cyclooxygenase (COX)-2 and COX-derived prostaglandin E2 (PGE<SUB>2</SUB>) production in lipopolysaccharide (LPS)-stimulated murine peritoneal macrophages. The compound also reduced tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) production and suppressed the phosphorylation and degradation of IκB-α and nuclear translocation of p65 in murine peritoneal macrophages in response to LPS stimulus. The inhibitory effects of pulchellamin G on nuclear factor kappa B (NF-κB) translocation was impaired by co-treatment of the cells with HO activity inhibitor tin protoporphyrin (SnPP). By using SnPP, we verified that the inhibitory effects of pulchellamin G on the pro-inflammatory mediators NO, PGE<SUB>2</SUB>, TNF-α, and IL-1β are associated with induction of HO-1 expression. Our data suggest that pulchellamin G might have potent therapeutic effects and it should be considered in the development of treatments for various inflammatory diseases.

Tetrachloroauric Acid Depresses the Activation Processes of Phagocytic Cells

Chung Soo Lee 대한생리학회-대한약리학회 1998 The Korean Journal of Physiology & Pharmacology Vol.2 No.3

<P> Gold compounds depress phagocytic cell responses, including chemotaxis, and respiratory burst. However, the effects of gold compounds on the function of phagocytic cells are variable according to the preparation of medicine. In this study, effect of tetrachloroauric acid on activated neutrophil responses, including respiratory burst, lysosomal enzyme release and change of intracellular Ca<SUP>2+</SUP> level and on the synthesis of interleukin-8 and granulocyte-macrophage colony stimulating factor by macrophages was studied. This study further examines how gold compounds affect the activation processes. <P> The respiratory burst stimulated by complement C5a, degraded IgG and PMA in neutrophils was inhibited by tetrachloroauric acid. In contrast to C5a and degraded IgG, PMA-stimulated superoxide production was weakly inhibited by tetrachloroauric acid. Staurosporine, genistein, EGTA and verapamil inhibited superoxide and H<SUB>2</SUB>O<SUB>2</SUB> production caused by C5a and degraded IgG. PMA-stimulated superoxide production was inhibited by staurosporine but was not affected by genistein. Tetrachloroauric acid, genistein, EGTA and verapamil inhibited the release of acid phosphatase and myeloperoxidase, while the effect of staurosporine was not detected. The synthesis of interleukin-8 and granulocyte-macrophage colony stimulating factor by interleukin-1β in macrophages was inhibited by tetrachloroauric acid. Preincubation with tetrachloroauric acid, genistein, EGTA and verapamil, the elevation of [Ca<SUP>2+</SUP>]<I><SUB>i</SUB></I> evoked by C5a was inhibited. Store-regulated Ca<SUP>2+</SUP> entry in thapsigargin-pretreated neutrophils was decreased by the addition of tetrachloroauric acid and genistein. The effect of staurosporine on intracellular Ca<SUP>2+</SUP> mobilization was not observed. In conclusion, tetrachloroauric acid may suppress neutrophil responses through its inhibitory action on elevation of intracellular Ca<SUP>2+</SUP> level and protein kinase C. It might exhibit an inhibitory effect on the action of protein tyrosine kinase. Tetrachloroauric acid depresses cytokine production by macrophages.

Immunohistochemical localization of galectin-3 in the brain with Theiler's murine encephalomyelitis virus (DA strain) infection

Shin, Taekyun,Carrillo-Salinas, Francisco J.,Martinez, Ana Feliu,Mecha, Miriam,Guaza, Carmen The Korean Society of Veterinary Science 2013 大韓獸醫學會誌 Vol.53 No.3

Galectin-3 is a ${\beta}$-galactoside-binding lectin that plays a role in neuroinflammation through cell migration, proliferation, and apoptosis. In the present study, regulation of galectin-3 was examined in the brain of mice infected with the Daniel strain of Theiler's murine encephalomyelitis virus (TMEV) at days 7 and 81 post-infection by immunohistochemistry. Immunohistochemistry revealed that galectin-3 was mainly localized in ionized calcium-binding adapter 1-positive macrophages/activated microglia, but not in Iba-1-positive ramified microglia. Galectin-3 was also weakly detected in some astrocytes in the same encephalitic lesions, but not in neurons and oligodendrocytes. Collectively, the present findings suggest that galectin-3, mainly produced by activated microglia/macrophages, may be involved in the pathogenesis of virus induced acute inflammation in the early stage as well as the chronic demyelinating lesions in Daniel strain of TMEV induced demyelination model.

Anti-Inflammatory and Anti-Superbacterial Activity of Polyphenols Isolated from Black Raspberry

Seong Keun Kim,Hyuna Kim,Song Ah Kim,Hee Kuk Park,Wonyong Kim 대한생리학회-대한약리학회 2013 The Korean Journal of Physiology & Pharmacology Vol.17 No.1

The fruit of the black raspberry (Rubus coreanus Miquel) has been employed in traditional medicine, and recent studies have demonstrated its measureable biological activities. However, the root of the black raspberry has not been studied. Therefore, in this study, we evaluated the anti-inflammatory and antibacterial properties of the root and unripe fruit polyphenols of the black raspberry. Both polyphenols proved to have anti-inflammatory activity as evidenced by the decreased nitric oxide (NO), cytokines (IL-1Ղ, IL-6, and IL-10) and prostaglandin E2 (PGE₂) levels in lipopolysaccharide (LPS)- stimulated RAW 264.7 murine macrophages. However, root polyphenols showed stronger anti- inflammatory activity than fruit polyphenols. LPS-induced mRNA and protein expressions of inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 levels were also decreased, confirming the anti- inflammatory activity. Root polyphenols showed lethal activity against methicillin-resistant Staphylococcus aureus (MRSA), carbapenem-resistant Acinetobacter baumannii (CRAB), and Bacillus anthracis. In contrast, the black raspberry fruit did not demonstrate these properties. These data provide the first demonstration that black raspberry root has potential anti-inflammatory and anti- superbacterial properties that can be exploited as alternatives for use in the food and cosmetic industries and/or as pharmaceuticals.