• Molecules and Cells
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• 제45권4호
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• pp.257-272
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• 2022

In addition to inducing apoptosis, caspase inhibition contributes to necroptosis and/or autophagy depending on the cell type and cellular context. In macrophages, necroptosis can be induced by co-treatment with Toll-like receptor (TLR) ligands (lipopolysaccharide [LPS] for TLR4 and polyinosinic-polycytidylic acid [poly I:C] for TLR3) and a cell-permeable pan-caspase inhibitor zVAD. Here, we elucidated the signaling pathways and molecular mechanisms of cell death. We showed that LPS/zVAD- and poly I:C/zVAD-induced cell death in bone marrow-derived macrophages (BMDMs) was inhibited by receptor-interacting protein kinase 1 (RIP1) inhibitor necrostatin-1 and autophagy inhibitor 3-methyladenine. Electron microscopic images displayed autophagosome/autolysosomes, and immunoblotting data revealed increased LC3II expression. Although zVAD did not affect LPS- or poly I:C-induced activation of IKK, JNK, and p38, it enhanced IRF3 and STAT1 activation as well as type I interferon (IFN) expression. In addition, zVAD inhibited ERK and Akt phosphorylation induced by LPS and poly I:C. Of note, zVAD-induced enhancement of the IRF3/IFN/STAT1 axis was abolished by necrostatin-1, while zVAD-induced inhibition of ERK and Akt was not. Our data further support the involvement of autocrine IFNs action in reactive oxygen species (ROS)-dependent necroptosis, LPS/zVAD-elicited ROS production was inhibited by necrostatin-1, neutralizing antibody of IFN receptor (IFNR) and JAK inhibitor AZD1480. Accordingly, both cell death and ROS production induced by TLR ligands plus zVAD were abrogated in STAT1 knockout macrophages. We conclude that enhanced TRIF-RIP1-dependent autocrine action of IFNβ, rather than inhibition of ERK or Akt, is involved in TLRs/zVAD-induced autophagic and necroptotic cell death via the JAK/STAT1/ROS pathway.

• 한국동물위생학회지
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• 제35권4호
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• pp.269-273
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• 2012

A confirmatory serological test, the standard tube agglutination test (STAT) is evaluated for the diagnostic efficiency in brucellosis Korea. A total of 345 bovine samples were collected from regional veterinary branch under national brucellosis monitoring program from January 2010 to June 2012 in Korea. These samples were diagnosed as suspected serum and brucellosis positive by the Rose Bengal test (RBT) and the STAT, respectively. The STAT was compared and evaluated with three serological test such as the indirect-enzyme linked immunosorbent assay (I-ELISA), competitive-enzyme linked immunosorbent assay (C-ELISA) and fluorescence polarisation assay (FPA) prescribed for international trade by OIE. Among the 345 bovine serum samples, 302 (87.5%) were diagnosed as positive in the STAT, while 215 (62.3%), 223 (64.6%) and 194 (56.2%) serum samples were diagnosed as positive for brucellosis in the I-ELISA, C-ELISA and FPA, respectively. The STAT showed quite high positive results as compared with three prescribed tests of OIE. FPA, I-ELISA and C-ELISA have shown 60.6%, 64.9% and 67.2% correlation, respectively as compared to the STAT. However correlations of three prescribed tests ranged high 84.1~97.7%. Especially, correlation between I-ELISA and C-ELISA is quite high, 97.7%. These results suggest that the STAT has shown many false-positive reactions. Therefore, additional serological test, such as ELISAs and FPA, would be necessary to adopt as a confirmatory test in the national surveillance program of bovine brucellosis in Korea.

• Natural Product Sciences
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• 제23권1호
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• pp.1-4
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• 2017

Sinensetin, a pentamethoxyflavone, is known to exert various pharmacological activities including anti-angiogenesis, anti-diabetic and anti-inflammatory activities. However, its effects on the human mast cell - 1 (HMC-1) mediated inflammatory mechanism remain unknown. To explore the mediator and cellular inflammatory response of sinensetin, we examined its influence on phorbol 12-myristate 13-acetate (PMA) plus A23187 induced inflammatory mediator production in a human mast cell line. In this study, interleukin (IL)-6 production was measured using the enzyme-linked immunosorbent assay and reverse transcription polymerase chain reaction. Sinensetin inhibited PMA plus A23187 induced IL-6 production in a dose-dependent manner as well as IL-4, IL-5 and IL-8 mRNA expression. Furthermore, sinensetin inhibited signal transducer and activator of transcription 3 (STAT3) phosphorylation, suggesting that sinensetin inhibits the production of inflammatory mediators by blocking STAT3 phosphorylation. Moreover, sinensetin was found to inhibit nuclear factor kappa B activation. These findings suggest that sinensetin may be involved in the regulation of mast cell-mediated inflammatory responses.

• Korean Journal of Acupuncture
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• 제24권2호
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• pp.129-139
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• 2007

목적 : Signal transducers and activators of transcription 6 (Stat 6) 유전자는 면역세포의 발달에 있어서 중요한 유전인자이며, IL-4와 같은 사이토카인에 의해 유전자 발현이 조절된다. 본 연구에서는 Stat 6 유전자 제거 생쥐와 정상 (wild type, W/T) 생쥐에 carrageenan으로 염증을 유도한 후 족삼리에 침치료를 시행하여 시상하부에서의 유전자 발현 양상을 분석하고자 하였다. 방 법 : BALB/c (W/T, n=12) and BALB/c-Stat 6 유전자 제거 생쥐 (n=12)의 발뒤꿈치 표피에 1% carrageenan을 30 ul 주사하여 염증을 유도하였다. 침은 염증 유도 30분 후에 족삼리(ST36)에 시침하였으며, 염증유도에 의한 부종 증가율을 매 시간마다 측정하여 총 5시간동안 측정하였다. 마이크로에러이는 Stat 6 유전자 제거 생쥐를 염증 유발 군과 염증유발 후 침을 처치한 군으로 나누고, 시상하부를 적출하여 RNA를 분리한뒤 마이크로어레이 프로파일을 분석하였다. 결 과 : 염증에 의한 부종증가율을 비교한 결과, Stat 6 유전자 제거 생쥐 그룹의 부종증가율이 W/T 생쥐의 부종 증가율보다 약 50 % 정도 감소하였으며, 각 3, 4, 5시간째에 유의한 차이를 나타내었다. (각 p<0.05). W.T생쥐군과 Stat 6 유전자 제거 생쥐군 모두에서, 침 처치군이 염증 유발 군에 비해, 염증 유발 2시간 후부터 유의한 감소를 나타내었다. 시상하부의 유전자 발현을 관찰한 결과, 39개의 유전자가 3배 이상 감소하였으며, 19개의 유전자는 3배 이상 증가하였다. 결 론 : W/T 생쥐군과 Stat 6 유전자 제거 생쥐 모두에서 침의 진통효과는 나타나며, 이의 기전에는 시상하부에서의 침 치료에 의한 염증관련 유전자들의 감소와, 항염증과 관련된 유전자들이 증가가 관여하는 것으로 보인다., 10, 11), 내측전완피신경(TE5, 6, 7, 8, 9, 10, 11), 후상완피신경(TE12, 13), 상외측상완피신경(TE13), 외측쇄골상신경(TE14, 15),대이개신경(TE16, 17, 18, 19), 소후두신경(TE19, 20), 이개측두신경(TE20, 21, 22), 안면신경측두지(TE22, 23), 관골측두신경(TE23), 중층에 견갑상신경(TE15), 견갑배신경(TE15), 경상설골근신경(TE17), 후이개신경(TE18, 19, 20), 안면신경측두지(TE20, 21, 22), 심층에 후골간신경(TE5, 6, 7), 요골신경심지(TE8, 9, 12, 13), 견갑상신경(TE14), 액와신경가지(TE14), 부신경(TE16), 안면신경과 부신경가지(TE17), 설인신경(TE17), 설하신경(TE17), 경신경고리(TE17), 미주신경(TE17), 안면신경 (TE18). 3) 혈(血) 관(管) : 천층에 척측정맥배측지(TE1, 2), 고유수장지동맥배측지(TE1), 배측중수골동맥배측지(TE2), 배측중수골정맥(TE3), 척측피정맥(TE4, 5, 6, 7, 8, 9, 10, 11), 배측정맥궁(TE4), 부요측피정맥(TE6, 8, 9),요측피정맥(TE10, 11), 후견봉정맥가지(TE13, 14), 후이개동 ${\cdot}$ 정맥(TE16, 17, 18, 19, 20), 전이개동 ${\cdot}$ 정맥(TE20), 천측두동 ${\cdot}$ 정맥(TE22, 23), 중층에 후상완회선동맥(TE14), 견갑배동맥(TE15), 견갑상동맥(TE15),천측두동 ${\cdot}$ 정맥(TE21), 관골측두동 ${\cdot}$ 정맥(TE23), 심층에 배측중수골동맥(TE3), 배측수근동맥궁(TE4), 후골간동맥(TE4, 5, 6, 7, 8, 9), 전골간동맥(TE6, 7, 9)

• Journal of Acupuncture Research
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• 제34권1호
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• pp.1-9
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• 2017

Objectives : We investigated the synergistic effects of bee venom (BV) and natural killer (NK) cells on B16F10 melanoma cell apoptosis mediated by IL-4. Methods : We performed a cell viability assay to determine whether BV can enhance the inhibitory effect of NK-92MI cells on the growth of B16F10 melanoma cells, and western blot analysis to detect changes in the expression of IL-4, $IL-4R{\alpha}$, and other apoptosis-related proteins. EMSA was performed to observe the activity of STAT6. To confirm that the inhibitory effect of BV and NK cells was mediated by IL-4, the above tests were repeated after IL-4 silencing by siRNA (50 nM). Results : B16F10 melanoma cells co-cultured with NK-92MI cells and simultaneously treated by BV ($5{\mu}g/ml$) showed a higher degree of proliferation inhibition than when treated by BV ($5{\mu}g/ml$) alone or co-cultured with NK-92MI cells alone. Expression of IL-4, $IL-4R{\alpha}$, and that of other pro-apoptotic proteins was also enhanced after co-culture with NK-92MI cells and simultaneous treatment with BV ($5{\mu}g/ml$). Furthermore, the expression of anti-apoptotic bcl-2 decreased, and the activity of STAT6, as well as the expression of STAT6 and p-STAT6 were enhanced. IL-4 silencing siRNA (50 nM) in B16F10 cells, the effects of BV treatment and NK-92MI co-culture were reversed. Conclusion : These results suggest that BV could be an effective alternative therapy for malignant melanoma by enhancing the cytotoxic and apoptotic effect of NK cells through an IL-4-mediated pathway.

• 한국약용작물학회지
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• 제18권1호
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• pp.1-8
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• 2010

Inonotus obliquus is one of the immune-regulatory substances and is recognized to play the role in the metabolic process of inflammation, allergy and immuntiy. The purpose of this study was to evaluate the effects of water extracts of Inonotus obliquus (IOW) on the liver lymphocyte immune function in the Sprague-Dawley male rats treated with carbon tetrachloride ($CCl_4$) to induce liver damage. Rats were fed with each experimental diet and water for 4 weeks. We found that effects of IOW on interferon-gamma (IFN-$\gamma$), signal transducer and activator of transcription 1 (STAT1), phospho-signal transducer and activator of transcription 1 (pSTAT1) and GATA-binding protein 3 (GATA-3) were decrease in vivo. Interleukin-4 (IL-4), STAT6, pSTAT6 and T-box expressed in T-cells (T-bet) decreased significantly lower in $CCl_4$+IOW group than the $CCl_4$ group. Our data indicated that cytokine protein production were increased in $CCl_4$ group and $CCl_4$+IOW group. As a result of this study, we assume that IOW fed could regulate the immuno-modulating functions through regulate the cytokine production capacity activated by liver damage.

• 한국정보과학회논문지:컴퓨팅의 실제 및 레터
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• 제11권4호
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• pp.312-333
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• 2005

SIP(Session Initiation Protocol)은 VoIP(Iroice over IP)와 같은 인터넷 멀티미디어 통신 시스템의 세션 제어 프로토콜로 제안되었으며 많은 표준안에서 채택하고 있다. SIP는 다양한 형태의 오디오와 비디오 형식과 품질이 보장된 실시간 데이터 전송을 지원하기 위해 복잡한 세션 제어 단계를 갖는다. 그러나 지금까지의 프로토콜 분석기는 SIP 기반 멀티미디어 통신 시스템에서 발생되는 트래픽을 상세하게 분석해주지 못하고 있다 이에 본 논문에서는 SIP 기반의 멀티미디어 통신 시스템의 세션 설정 단계에서 데이터 교환 및 세션 변경에 따른 세부 정보를 분석하고 진단할 수 있는 도구로 새로운 프로토콜 분석기를 설계하고 구현한다. 제안된 프로토콜 분석기는 STAT(SIP based Traffic Analysis Tool)이라고 부르며 범용성과 확장성이 있도록 PC Windows에서 사용 가능한 소프트웨어로 구현되었다. STAT는 이더넷의 특징인 동보 기능을 이용하여 저수준 패킷을 수집하여 분석함으로써 SIP 기반 통신 시스템에 직접적인 영향 없이 세션 연결 상태와 실시간 트래픽 모니터링 정보를 제공한다. 따라서 본 논문에서 구현한 STAT는 SIP 기반 멀티미디어 통신 시스템의 개발 및 관리 도구로 활용될 수 있다.

• 대한본초학회지
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• 제27권4호
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• pp.9-16
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• 2012

Objective : Canavalia gladiata DC semen (CGS) have been used to improve hematopoietic activity. In the current study, we investigated whether CGS regulate hemato-potentiating function using hematopoietic stem cells (HSCs) as a testing system. Methods : HSCs isolated from femur in mice with leukopenia and thrombocytopenia induced induced by CTX. Then, Real-time PCR was performed to measure the mRNA expression and hematopoietic related gene (EPO, IL-3, SCF, c-kit, GM-CSF), the phoaphorylation of GATA-1 and STAT-5a/b were observed by ELISA method, and the number of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E), semisolid clonogenic assay was performed. Result : When HSCs were treated with CGS, the expression of hematopoietic related genes (EPO, IL-3, SCF, c-kit, and GM-CSF) were significantly increased at the levels of mRNA as well as production in HSCs. Additionally, CGS enhanced phosphorylation of STAT-1 and signal transducer and activator of transcription-5a/b (STAT-5a/b) in HSCs. Furthermore, CGS significantly enhanced the growth rate of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E) in vitro. Conclusion : These result suggest that CGS has hematopoietic enhancement via hematopoietic cytokine-mediated GATA-1/STAT-5a/b pathway.

• Biomolecules & Therapeutics
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• 제16권4호
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• pp.394-402
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• 2008

Atopic dermatitis (AD) is an inflammatory skin disease commonly characterized by infiltration of inflammatory cells into skin lesions. Keratinocytes produce many chemokines that are involved in the pathogenesis of skin disorders. In particular, macrophage-derived chemokine (MDC/CCL22) and thymus and activationregulated chemokine (TARC/CCL17) are Th2-type cytokines. Serum MDC and TARC levels are increased in AD patients. In this study, we investigated the anti-inflammatory effect and mechanism of action of the active fraction from Prunus yedoensis bark. We evaluated their inhibitory effects on the AD-like inflammatory markers (MDC and TARC) and JAK-STAT pathway (STAT1) in HaCaT keratinocytes. The EtOAc fraction of the crude extract (80% EtOH) and the E5 sub-fraction potently inhibited the induction of MDC and TARC mRNA and protein at 50 ${\mu}g$/mL in HaCaT cells. In addition, the E5 sub-fraction inhibited the phosphorylation of STAT1 protein associated with IFN-$\gamma$ signaling transduction in a dose-dependent manner. Thus, P. yedoensis may have antiatopic activity by suppressing the inflammatory chemokines (MDC and TARC).

• 한국식품과학회지
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• 제48권1호
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• pp.66-71
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• 2016

델피니딘은 양전하를 뛰는 diphenylpropane의 polyphenolic ring 구조를 가진 주요한 안토시아닌 색소 중에 하나이다. 최근 연구에서 델피니딘은 항산화, 항염증 뿐만 아니라 항암 효능을 가진다고 보고되었다. 본 연구에서는 전립샘 암에서 종양의 성장과 신생혈관생성에 관련된 중요한 인자인 VEGF 발현에 대한 델피니딘의 억제 효과를 조사하였다. RT-PCR을 통해 델피니딘을 처리한 PC3M 전립샘 암세포 세포에서 EGF로 유도한 VEGF mRNA 발현 수준이 감소됨을 확인하였다. 또한 델피니딘은 VEGF의 전사인자인 HIF-$1{\alpha}$와 STAT3가 세포 핵으로 전위되는 것을 효과적으로 억제하였다. 한편 luciferase assay을 통해 HRE-promoter 활성을 확인해 본 결과, 델피니딘이 HIF-$1{\alpha}$의 전사 활성을 억제시켜 VEGF 발현을 감소시키는 것을 알 수 있었다. 그리고 델피니딘은 EGFR의 발현에는 영향을 미치지 않고, Akt, p70S6K, 4EBP1의 인산화를 특이적으로 억제하는 것으로 나타났다. 결론적으로 델피니딘이 HIF-$1{\alpha}$와 STAT3, VEGF 발현을 억제를 통하여 암세포 증식억제와 신생혈관형성을 억제하는 역할을 새롭게 확인하였다.