국소뇌허혈 후 허혈경계영역에서의 Activating Transcription Factor 3의 발현

송대용(Dae-Yong Song),오경민(Kyoung-Min OH),이지혜(Ji-Hye Lee),우란숙(Ran-Sook Woo),이윤정(Yun-Jeong Lee),한정태(Jung-Tae Han),백태경(Tai-Kyoung Baik) 대한해부학회 2008 Anatomy & Cell Biology Vol.41 No.3

c-Jun이나 fos와 같은 immediate early gene (IEGs)은 여러종류의 세포손상에 대해 즉각적으로 발현하여 손상 받은 신경세포의 운명을 결정함에 중요한 역할을 담당하는 것으로 알려져 있다. IEGs 중 activating transcription factor 3 (ATF3)는 다양한 신경손상에 반응하여 발현되며, 손상된 신경세포의 내∙외적 환경에 따라 신경세포의 생존 혹은 신경세포의 사멸에도 관여할 수 있기 때문에 매우 흥미로운 단백질이다. 그러나 아직까지 뇌허혈손상에서 신경세포내 ATF3의 발현 및 역할에 대한 연구는 매우 드물다. 이에 이 연구는 중간대뇌동맥 폐색-재관류로 야기된 국소적 뇌허혈손상에서 초래되는 신경조직의 형태적 변화와 신경세포에서 일어나는 ATF3 발현의 변화를 조사하기 위하여 시도되었다. 허혈-재관류손상이 야기된 흰쥐의 뇌를 절취한 후 2mm 간격으로 연속 관상절편을 제작하고 triphenyltetrazolium chloride (TTC) 염색을 시행하여 중간대뇌동맥 폐색-재관류에 따른 뇌손상의 범위를 확인하였으며, 허혈중심영역 (ischemic core region) 및 허혈경계영역 (ischemic penumbra region)으로 구분하여 관찰하였다. Haematoxylin 및 eosin 염색결과 양 영역 모두에서 핵농축과 호염기성 신경세포체 변성과 같은 세포사멸을 암시하는 소견을 나타내는 신경세포를 다수 확인할 수 있었으며, 또한 많은 수의 아교세포가 동원된 소견을 제시할 수 있었다. 동원된 아교세포의 종류 를 동정하기 위해 GFAP 및 OX42에 대한 항체를 이용한 면역조직화학염색을 실시한 결과, 활성 별아교세포 및 미세아교세포가 뇌손상부위에 운집되어 있었다. 그리고 ATF3 면역조직화학염색을 실시한 결과, ATF3에 면역양성반응을 보이는 신경세포들은 허혈-재관류에 의한 손상이 야기된 대뇌겉질에서만 관찰되었으며, 대조군으로 사용한 반대측 대뇌겉질에서는 관찰되지 않았다. 특히 ATF3 면역양성세포는 허혈중심영역보다 허혈경계영역에서 더 빈번히 관찰되었다. 허혈중심영역에서의 신경세포사멸은 주로 세포괴사(necrosis)에 의해, 허혈경계영역에서의 세포사멸은 주로 세포자멸사(apoptosis)에 의해 유도된다는 여러 학자들의 보고를 참고하면 이번 연구결과는 ATF3가 허혈에 의해 야기되는 세포자멸사 혹은 세포생존 신호전달과정에 관여하여 세포의 운명을 결정하는 매우 중요한 IEGs 중 하나일 것임을 강력히 시사한다. It has been demonstrated that some of immediate early genes (IEGs) such as c-Jun or fos are induced immediately following neuronal injury and they play an important role in determining the fate of the injured neurons. Of IEGs, the activating transcription factor 3 (ATF3) is focused by many investigators, because they are expressed in various types of neural insults and have been known to serve a diverse function in both neuronal survival and death. However, little is known about the functional role of ATF3 in ischemic brain injury. So in this study, the authors examined the expression pattern of the activating transcription factor 3 (ATF3) following middle cerebral artery (MCA) occlusion-reperfusion injury. According to the findings obtained by triphenyltetrazolium chloride (TTC) stains, the authors have classified the infarcted area into two regions, the ischemic core region and the ischemic penumbra region. In both regions, many neurons underwent neuronal degeneration, characterized by the shrunken nuclei with eosinophilic perikaryon. The H & E stain also demonstrated the increased number of probable activated astrocytes and microglia in the ischemic brain regions and this was confirmed by GFAP- and OX42-immunohistochemistry. Immunohistochemical study for ATF3 also demonstrated the specific upregulation of ATF3 in the nuclei of neurons under ischemic injury, but not in those of the contralateral regions. Interestingly, the number of the ATF3 positive neurons in the ischemic penumbra regions outnumbered that of the ischemic core regions. Based on many reports that the neuronal death in ischemic penumbra region is caused by programed cell death rather than by necrosis which is main cause of neuronal death in ischemic core region, our results could suggest that the ATF3 is an important IEGs which determine the fate of the ischemic neurons.

ATF3 Confers Resistance to Pneumococcal Infection Through Positive Regulation of Cytokine Production

Nguyen, Cuong Thach,Kim, Eun-Hye,Luong, Truc Thanh,Pyo, Suhkneung,Rhee, Dong-Kwon Oxford University Press 2014 The Journal of Infectious Diseases Vol.210 No.11

<P><B><I>Background.</I></B> Activating transcription factor–3 (ATF3) is known as a suppressor of cytokine production after exposure to lipopolysaccharide or during gram-negative bacterial infection. However, the mechanism by which ATF3 regulates innate immunity against gram-positive bacterial infection, particularly <I>Streptococcus pneumoniae</I>, remains unknown.</P><P><B><I>Methods.</I></B> The wild-type and ATF3 knock-out (KO) mice were infected intranasally (<I>i.n</I>) or intraperitoneally with <I>S. pneumoniae</I>, and bacterial colonization or survival rate was determined. Pneumococcal pneumonia was induced by <I>i.n</I> infection, and ATF3 level was determined by Western blot. ATF3 KO cells or ATF3 siRNA transfection were used to determine expression of ATF3 downstream genes. Enzyme-linked immunosorbent assay was used to examine cytokines levels.</P><P><B><I>Results.</I></B> ATF3 was highly expressed in various cell lines in vitro and in many organs in vivo. Pneumolysin was a novel inducer of ATF3. Pneumococcal infection induced ATF3, which subsequently stimulated production of cytokines (tumor necrosis factor [TNF]–α, interleukin [IL]–1β, and interferon [IFN]–γ). ATF3-mediated cytokine induction protected the host from pneumococcal infection. In the pneumonia infection model, the bacterial clearance of wild-type mice was more efficient than those of ATF3 KO mice.</P><P><B><I>Conclusions.</I></B> Taken together, we can conclude that ATF3 regulates innate immunity positively upon pneumococcus infection by enhancing TNF-α, IL-1β, and IFN-γ expression and modulating bacterial clearance.</P>

TLR4 Mediates Pneumolysin-Induced ATF3 Expression through the JNK/p38 Pathway in Streptococcus pneumoniae-Infected RAW 264.7 Cells

Nguyen, Cuong Thach,Kim, Eun-Hye,Luong, Truc Thanh,Pyo, Suhkneung,Rhee, Dong-Kwon Korean Society for Molecular and Cellular Biology 2015 Molecules and cells Vol.38 No.1

Activating transcription factor-3 (ATF3) acts as a negative regulator of cytokine production during Gram-negative bacterial infection. A recent study reported that ATF3 provides protection from Streptococcus pneumoniae infection by activating cytokines. However, the mechanism by which S. pneumoniae induces ATF3 after infection is still unknown. In this study, we show that ATF3 was upregulated via Toll-like receptor (TLR) pathways in response to S. pneumoniae infection in vitro. Induction was mediated by TLR4 and TLR2, which are in the TLR family. The expression of ATF3 was induced by pneumolysin (PLY), a potent pneumococcal virulence factor, via the TLR4 pathway. Furthermore, ATF3 induction is mediated by p38 mitogen-activated protein kinase (MAPK) and c-Jun N-terminal kinase (JNK). Thus, this study reveals a potential role of PLY in modulating ATF3 expression, which is required for the regulation of immune responses against pneumococcal infection in macrophages.

트레드밀 운동이 만성 신경통 동물 모델의 통증지표 및 통증 회피 반응에 미치는 영향

박재성 인문사회과학기술융합학회 2018 예술인문사회융합멀티미디어논문지 Vol.8 No.6

만성 신경통은 말초신경 손상 및 중추신경을 포함한 다양한 원인에 의해 발생되는 병리생리학적으로 복잡한 임상증상이다. 만성 신경통의 대응 및 치료방법은 의사, 치료사 등 의료진들과 환자 및 보호자들에게도 풀어야할 커다란 문제로 남아있다. 최근 선행연구들을 통해 몇 가지 동물 통증 모델들이 수영 및 유산소성 운동에 의해 완화되는 것을 보고한 바 있다. 하지만 아직까지 신경손상에 의한 만성 수축성 신경통(CCI)에 대한 연구 그리고 운동이 만성 수축성 신경통(CCI) 동물 모델의 어떠한 경로를 통해 신경통 완화에 미치는지 세포수준의 연구는 미진한 실정이다. 이에 본 연구자는 동물 실험을 통해 신경손상에 의한 만성 수축성 신경통 모델의 유산소성 운동이 통증에 미치는 영향을 알아보고자 하였다. 이를 위해 실험동물에 신경손상 수술을 시행하여 인위적으로 만성 수축성 신경통을 유발하였고, 이 동물 모델에 주 3회 저강도 유산소성 운동을 수행한 후 통증 발현 시 나타나는 생체반응 및 지표들을 확인 및 비교하여 가설을 검증하였다. 본 실험에서 확인한 통증 생체반응은 감각신경 활동 전위(Sensory Nerve Action Potential: SNAP)와 열 통증 회피반응(Heat Withdrawal Latency) 검사를 수행하였다. 또한 통증 발현 시 나타나는 생체지표로 알려진 ATF-3 발현량을 비교 분석하였다. 본 실험을 통해 얻어진 결과에 따르면 유산소성 운동을 수행한 집단이 운동을 수행하지 않은 집단보다 전달되는 감각신경 활동전위의 양도 많고 전달 속도역시 빠른 것으로 나타났다. 또한 인체 내에서 통증 발현 시 나타나는 것으로 알려진 생체통증지표 ATF-3 발현량이 운동을 수행한 집단에서 운동을 수행하지 않은 집단에 비해 낮게 나타나는 것을 확인하였다. 마지막으로 동물의 발바닥에 열통증 회피반응을 검사한 결과 운동을 수행한 집단에서 운동을 수행하지 않은 집단보다 빠른 회피반응을 나타내는 것을 확인하였다. 이는 저강도 유산소성 운동이 신경손상에 의한 만성 수축성 신경통에 긍정적인 효과를 나타내는 것으로 결론지을 수 있다. Chronic Peripheral neuropathy is a major complication of many clinical and therapeutic agents. Even though, many people try to fine the effective methods to prevent or stop developing it, but currently there is no effective methods found. Recent studies report that the swimming exercise attenuates neuropathic pain that were caused by diabetes. But the neuropathy caused by peripheral nerve injury is still unclear. In the present study, we used the model of Chronic Constrictive Injury(CCI) animal model that are widely used for chronic neuropathic pain. For this study, we made Chronic Constructive Injury(CCI) mouse model with cut and suture the sciatic nerve surgery. And we applied the low intensity of aerobic exercise for 3 days per week and compared the pain avoidance ability and pain bio-markers with no exercise control group. To detect the sensory functions and pain avoidance ability, we used the pain avoidance test with heat withdrawal latency tester and Sensory nerve action potential: SNAP). And we had compared and analyzed the ATF-3 that are known as a pain bio-marker. The results said that the low intensity of aerobic exercise on CCI model group enhance the recovery of Sensory nerve action potential(SNAP) than no exercised control group. Also we could detect the lower expression level of ATF-3 known as a pain bio-marker in the lower exercised group than no exercised control group. And we could detect faster reaction in the low exercised group than no exercise control group in the heat shock reaction(heat withdrawal latency) test. This means that the aerobic exercise group has analgesic effect and has better pain reaction than no exercise control group. We had concluded that the analgesic effect of exercise on the CCI animal model with the ATF-3 expression level and behavior test result. This means that the low intensity of aerobic exercise has beneficial effect on Chronic Constrictive Injury(CCI).

TLR4 Mediates Pneumolysin-Induced ATF3 Expression through the JNK/p38 Pathway in Streptococcus pneumoniae-Infected RAW 264.7 Cells

Cuong Thach Nguyen,이동권,김은혜,Truc Thanh Luong,표석능 한국분자세포생물학회 2015 Molecules and cells Vol.38 No.1

Activating transcription factor-3 (ATF3) acts as a negative regulator of cytokine production during Gram-negative bacterial infection. A recent study reported that ATF3 provides protection from Streptococcus pneumoniae infection by activating cytokines. However, the mechanism by which S. pneumoniae induces ATF3 after infection is still unknown. In this study, we show that ATF3 was upregulated via Toll-like receptor (TLR) pathways in response to S. pneumoniae infection in vitro. Induction was mediated by TLR4 and TLR2, which are in the TLR family. The expression of ATF3 was induced by pneumolysin (PLY), a potent pneumococcal virulence factor, via the TLR4 pathway. Furthermore, ATF3 induction is mediated by p38 mitogen-activated protein kinase (MAPK) and c-Jun N-terminal kinase (JNK). Thus, this study reveals a potential role of PLY in modulating ATF3 expression, which is required for the regulation of immune responses against pneumococcal infection in macrophages.

ER Stress-Inducible ATF3 Suppresses BMP2-Induced ALP Expression and Activation in MC3T3-E1 Cells

Jae-kyung Park,Hoon Jang,Donghwan Kim,Wu-sheng Sun,Hyoung-Joo Kim,Jeong-Woong Lee 한국동물생명공학회(구 한국동물번식학회) 2014 한국동물번식학회 한중일 심포지엄 Vol.2014 No.1

Larval hemolymph of rhinoceros beetle, <i>Allomyrina dichotoma</i>, enhances insulin secretion through ATF3 gene expression in INS-1 pancreatic β-cells

Kim, Seung-Whan,Suh, Hyun-Woo,Yoo, Bo-Kyung,Kwon, Kisang,Yu, Kweon,Choi, Ji-Young,Kwon, O-Yu Verlag der Zeitschrift fuer Naturforschung 2018 Zeitschrift fur Naturforschung. Section C Vol.73 No.9

<P><B>Abstract</B></P><P>In this study, we show that INS-1 pancreatic β-cells treated for 2 h with hemolymph of larvae of rhinoceros beetle,<I>Allomyrina dichotoma</I>, secreted about twice as much insulin compared to control cells without such treatment. Activating transcription factor 3 (ATF3) was the highest upregulated gene in DNA chip analysis. The<I>A. dichotoma</I>hemolymph dose-dependently induced increased expression levels of genes encoding ATF3 and insulin. Conversely, treatment with ATF3 siRNA inhibited expression levels of both genes and curbed insulin secretion. These results suggest that the<I>A. dichotoma</I>hemolymph has potential for treating and preventing diabetes or diabetes-related complications.</P>

6-Hydroxydopamine induces nuclear translocation of apoptosis inducing factor in nigral dopaminergic neurons in rat

Hong-Il Yoo,Gil-Yeong Ahn,이은진,Eu-gene Kim,Sung-Young Hong,Sang-Jin Park,Ran-Sook Woo,Tai-Kyoung Baik,Dae-Yong Song,T.-K. Baik,D.-Y. Song 대한독성 유전단백체 학회 2017 Molecular & cellular toxicology Vol.13 No.3

Parkinson’s disease (PD) is a progressive neurodegenerative disorder, characterized by relatively selective death of dopaminergic (DA) neurons in the substantia nigra pars compacta (SNpc). 6-Hydroxydopamine (6-OHDA), a neurotoxin that causes the death of DA neurons, is commonly used to produce experimental PD model in rodents. Accumulating evidences suggest that caspase-independent apoptotic programmed cell death (PCD) could also be involved in the progression of various neurodegenerative diseases in addition to caspase-dependent neuronal PCD. Apoptosis-inducing factor (AIF), a mitochondrial intermembrane oxidoreductase, has been identified as a key protein implicated in caspase-independent apoptosis. However, little is known about the role of AIF in death of nigral DA neurons in PD. Therefore, we undertook this study in an effort to clarify the involvement of AIF in DA neuronal death by 6-OHDA administration. Ten and twenty micrograms of 6-OHDA was infused into the medial forebrain bundle (MFB) unilaterally, and the experimental rats were sacrificed at various time point. The DA neuronal loss was identified in the ipsilateral SN in the dose-dependent manner by using NeuN and tyrosine hydroxylase immunohistochemical staining and western blot assay. Numerous degenerating neurons, showing apoptotic features which are characterized by the shrunken nuclei with eosinophilic perikarya were observed in the ipsilateral SNpc. Activating transcription factor 3 (ATF3), the specific marker for neuronal damage, was expressed in the ipsilateral DA neurons only. Immunohistochemistry and immunofluorescence staining demonstrated that nuclear localization of AIF in the ipsilateral degenerating DA neurons. These results suggest that AIF could induce DA neuronal death by caspase-independent apoptosis in 6-OHDA treated model, although other cell death cascades should not be rule out.

Acquisition of Chemoresistance and Other Malignancy-related Features of Colorectal Cancer Cells Are Incremented by Ribosome-inactivating Stress

Oh, Chang-Kyu,Lee, Seung Joon,Park, Seong-Hwan,Moon, Yuseok American Society for Biochemistry and Molecular Bi 2016 The Journal of biological chemistry Vol.291 No.19

<P>Colorectal cancer (CRC) as an environmental disease is largely influenced by accumulated epithelial stress from diverse environmental causes. We are exposed to ribosome-related insults, including ribosome-inactivating stress (RIS), from the environment, dietary factors, and medicines, but their physiological impacts on the chemotherapy of CRC are not yet understood. Here we revealed the effects of RIS on chemosensitivity and other malignancy-related properties of CRC cells. First, RIS led to bidirectional inhibition of p53-macrophage inhibitory cytokine 1 (MIC-1)-mediated death responses in response to anticancer drugs by either enhancing ATF3-linked antiapoptotic signaling or intrinsically inhibiting MIC-1 and p53 expression, regardless of ATF3. Second, RIS enhanced the epithelial-mesenchymal transition and biogenesis of cancer stem-like cells in an ATF3-dependent manner. These findings indicate that gastrointestinal exposure to RIS interferes with the efficacy of chemotherapeutics, mechanistically implying that ATF3-linked malignancy and chemoresistance can be novel therapeutic targets for the treatment of environmentally aggravated cancers.</P>

Early growth response protein 1 upregulation and nuclear translocation by 2'-benzoyloxycinnamaldehyde induces prostate cancer cell death

Kang, H.S.,Ock, J.,Lee, H.J.,Lee, Y.J.,Kwon, B.M.,Hong, S.H. Elsevier ; Elsevier Science Pub. Co 2013 Cancer letters Vol.329 No.2

2'-Benzoyloxycinnamaldehyde (BCA) induces apoptosis in human cancer cells through ROS generation. BCA upregulates proapoptotic genes such as activating transcription factor 3 (ATF3), NSAID-activated gene 1 protein (NAG-1), and growth arrest and DNA-damage-inducible protein alpha (GADD45A) in prostate cancer cells. These genes are known to be induced by transcription factor early growth response protein 1 (EGR1). BCA induces significant EGR1 upregulation, while EGR1 knockdown decreases the induction of these genes with concurrent alleviation of cell death by BCA. Antioxidant glutathione pretreatment with BCA removes EGR1 expression increase, suggesting that EGR1 upregulation is dependent on oxidative stress generated by BCA. In prostate cancer cells, EGR1 localizes in the cytoplasm; however, BCA remarkably upregulates EGR1 nuclear translocalization, suggesting its possible effect as a transcriptional activator. BCA induces transient upregulation of importin-7 (IPO7) which is critical for EGR1 nuclear translocation, and IPO7 knockdown led to a significant decrease in chemosensitivity to BCA. Taken together, our findings suggest that BCA induces prostate cancer cell death via EGR1 upregulation and nuclear translocalization, followed by activation of proapoptotic target genes.