스트레스-유도 열충격단백질 27(Heat Shock Protein 27)의 활성과 물리치료의 상관성

김미선,이성호,김일현,황병용,김중환,Kim, Mi-Sun,Lee, Sung-Ho,Kim, Il-Hyun,Hwang, Byong-Yong,Kim, Jung-Hwan 대한물리치료학회 2008 대한물리치료학회지 Vol.20 No.1

Purpose: Heat shock proteins (HSPs) are a group of proteins that are activated when cells are exposed to a variety of environmental stresses, such as infection, inflammation, exposure to toxins, starvation, hypoxia, brain injury, or water deprivation. The activation of HSPs by environmental stress plays a key role in signal transduction, including cytoprotection, molecular chaperone, anti-apoptotic effect, and anti-aging effects. However, the precise mechanism for the action of small HSPs, such as HSP27 and mitogen-activated protein kinases (MAPKs: extracellular-regulated protein kinase 1/2 (ERK1/2), p38MAPK, stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK), is not completely understood, particularly in application of cell stimulators including platelet-derived growth factor (PDGF), angiotensin II (AngII), tumor necrosis factor $\alpha$ (TNF$\alpha$), and $H_2O_2$. This study examined the relationship between stimulators-induced enzymatic activity of HSP27 and MAPKs from rat smooth and skeletal muscles. Methods: 2-dimensional electrophoresis (2DE) and matrix assisted laser desorption ionizationtime-of-flight/time-of-flight (MALDI-TOF/TOF) analysis were used to identify HSP27 from the intact vascular smooth and skeletal muscles. Three isoforms of HSP27 were detected on silver-stained gels of the whole protein extracts from the rat aortic smooth and skeletal muscle strips. Results: The expression of PDGF, AngII, TNF$\alpha$, and $H_2O_2$-induced activation of HSP27, p38MAPK, ERK1/2, and SAPK/JNK was higher in the smooth muscle cells than the control. SB203580 (30${\mu}$M), a p38MAPK inhibitor, increased the level of HSP27 phosphorylation induced by stimulators in smooth muscle cells. Furthermore, the age-related and starvation-induced activation of HSP27 was higher in skeletal muscle cells (L6 myoblast cell lines) and muscle strips than the control. Conclusion: These results suggest, in part, that the activity of HSP27 and MAPKs affect stressors, such as PDGF, AngII, TNF$\alpha$, $H_2O_2$, and starvation in rat smooth and skeletal muscles. However, more systemic research will be needed into physical therapy, including thermotherapy, electrotherapy, radiotherapy and others.

p38 Mitogen-Activated Protein Kinase is Involved in Endoplasmic Reticulum Stress-Induced Cell Death and Autophagy in Human Gingival Fibroblasts

Kim, Do-Sung,Kim, Jung-Hoon,Lee, Geum-Hwa,Kim, Hyung-Tae,Lim, Jung Min,Chae, Soo-Wan,Chae, Han-Jung,Kim, Hyung-Ryong Pharmaceutical Society of Japan 2010 Biological & pharmaceutical bulletin Vol.33 No.4

<P>Inflammation or hypoxia in gingival tissue can induce endoplasmic reticulum (ER) stress, which is related with autophagy. The autophagy is a catabolic process involving the degradation of a cell's own components. Although autophagy resulting in the total destruction of the cell is one of cell death types, no conclusive evidence exists for such a process. In order to examine the association of ER stress and autophagy in gingival system, ER stress agents brefeldin A, thapsigargin, and tunicamycin were exposed to human gingival cells. The ER stress agents induced cell death and the expression of ER stress proteins, glucose-regulated protein of 78 kDa (GRP78) and CCAAT/enhancer-binding protein (C/EBP) homologous protein (CHOP). ER stress also increased the formation of autophagic vesicles with the expression of beclin and LC-3 (microtubule-associated protein1 light chain 3) II, two autophagic markers. ER stress induced the phosphorylation of p38MAPK (mitogen-activated protein kinase), and the p38MPAK inhibitor, SB203580, inhibited the resulting cell death and autophagy. In summary, ER stress induces cell death and autophagy through p38MAPK in human gingival cells.</P>

SB202190- and SB203580-Sensitive p38 Mitogen-Activated Protein Kinase Positively Regulates Heat Shock- and Amino Acid Analog-Induced Heat Shock Protein Expression

Sun Hee Kim,Song Iy Han,Su Young Oh,Myoung Suk Seo,Hye Gyeong Park,Ho Sung Kang 대한의생명과학회 2003 Biomedical Science Letters Vol.9 No.2

When cells are exposed to proteotoxic stresses such as heat shock, amino acid analogs, and heavy metals, they increase the synthesis of the heat shock proteins (HSPs) by activating the heat shock transcription factor 1 (HSF1), whose activity is controlled via multiple steps including homotrimerization, nuclear translocation, DNA binding, and hyperphosphorylation. Under unstressed conditions, the HSF1 activity is repressed through its constitutive phosphorylation by glycogen synthase kinase 3β (GSK3β), extracellular regulated kinase ½ (ERK½), and stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK). However, the protein kinase (s) responsible for HSF1 hyperphosphorylation and activation is not yet identified. In the present study, we observed that profile of p38 mitogen-activated protein kinase (p38MAPK) activation in response to heat shock was very similar to those of HSF1 hyperphosphorylation and nuclear translocation. Therefore, we investigated whether p38MAPK is involved in the heat shock-induced HSF1 activation and HSP expression. Here we show that the p38MAPK inhibitors, SB202190 and SB203580, but not other inhibitors including the MEK½ inhibitor PD98059 and the PI3-K inhibitor LY294002 and wortmannin, suppress HSF1 hyperphosphorylation in response to heat shock and L-azetidine 2-carboxylic acid (Azc), but not to heavy metals. Furthermore, heat shock-induced HSF1-DNA binding and HSP72 expression was specifically prevented by the p38MAPK inhibitors, but not by the MEK½ inhibitor and the PI3-K inhibitors. These results suggest that SB202190- and SB203580-sensitive p38MAPK may positively regulate HSP gene regulation in response to heat shock and amino acid analogs.

SB202190- and SB203580-Sensitive p38 Mitogen-Activated Protein Kinase Positively Regulates Heat Shock- and Amino Acid Analog-Induced Heat Shock Protein Expression

Kim, Sun-Hee,Han, Song-Iy,Oh, Su-Young,Seo, Myoung-Suk,Park, Hye-Gyeong,Kang, Ho-Sung The Korean Society for Biomedical Laboratory Scien 2003 Journal of biomedical laboratory sciences Vol.9 No.2

When cells are exposed to proteotoxic stresses such as heat shock, amino acid analogs, and heavy metals, they increase the synthesis of the heat shock proteins (HSPs) by activating the heat shock transcription factor 1 (HSF1), whose activity is controlled via multiple steps including homotrimerization, nuclear translocation, DNA binding, and hyperphosphorylation. Under unstressed conditions, the HSF1 activity is repressed through its constitutive phosphorylation by glycogen synthase kinase 3$\beta$ (GSK3$\beta$), extracellular regulated kinase 1/2 (ERK1/2), and stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK). However, the protein kinase (s) responsible for HSF1 hyperphosphorylation and activation is not yet identified. In the present study, we observed that profile of p38 mitogen-activated protein kinase (p38MAPK) activation in response to heat shock was very similar to those of HSF1 hyperphosphorylation and nuclear translocation. Therefore, we investigated whether p38MAPK is involved in the heat shock-induced HSF1 activation and HSP expression. Here we show that the p38MAPK inhibitors, SB202190 and SB203580, but not other inhibitors including the MEK1/2 inhibitor PD98059 and the PI3-K inhibitor LY294002 and wortmannin, suppress HSF1 hyperphosphorylation in response to heat shock and L-azetidine 2-carboxylic acid (Azc), but not to heavy metals. Furthermore, heat shock-induced HSF1-DNA binding and HSP72 expression was specifically prevented by the p38MAPK inhibitors, but not by the MEK1/2 inhibitor and the PI3-K inhibitors. These results suggest that SB202190- and SB203580-sensitive p38MAPK may positively regulate HSP gene regulation in response to heat shock and amino acid analogs.

Effects of Mining-impacted Polished Rice on the Glutathione Peroxide and Glutathione S-transferase Activities, and Oxidative Stress-induced Apoptosis Signal Regulating Kinase 1, p38 Mitogen-activated Protein Kinase Isomer, and Jun N-terminal Kinase in Hu

Ji-Ho Lee,Eun-Jung Jeong,Geon-Jae Im,Won-Il Kim 한국응용생명화학회 2012 Applied Biological Chemistry (Appl Biol Chem) Vol.55 No.2

In toxic elements (TEs)-contaminated polished rice,human keratinocytes cell viability was significantly inhibited to a maximum of 30%. Glutathione peroxide (GPx) and glutathione Stransferase (GST) activities, and gene activation of apoptosis signal regulating kinase 1 (ASK1) were significantly induced at 72 h. Strong activation in p38α-, weak activation in p38δ-, and no expression in p38 β,γ-mitogen-activated protein kinase (MAPK),and jun N-terminal kinase (JNK) were observed at 24 and 72 h. At protein level, the ASK1 and p38 MAPK activities were found to be strongly stimulated at 72 h, whereas phosphorylated ASK1 and p38 MAPK proteins were not detected in contaminated polished rice. Both JNK and phosphorylated JNK were not activated at 24and 72 h. These results demonstrate that mine-impacted polished rice induced HaCaT cell toxicity associated with oxidative stress.

Nitric Oxide로 유발된 아포토시스에 대한 Sek 1의 영향

오재민,박래길,소흥섭,문연자,백용아,최민규,박옥규,이호익,정연태 대한해부학회 1999 Anatomy & Cell Biology Vol.32 No.5

ER Stress Is Implicated in Mitochondrial Dysfunction-Induced Apoptosis of Pancreatic Beta Cells

June Woo Lee,Won Ho Kim,여지영,정명호 한국분자세포생물학회 2010 Molecules and cells Vol.30 No.6

Mitochondrial dysfunction induces apoptosis of pancreatic β-cells and leads to type 2 diabetes, but the mechanism involved in this process remains unclear. Chronic endoplasmic reticulum (ER) stress plays a role in the apoptosis of pancreatic β-cells; therefore, in current study, we investigated the implication of ER stress in mitochondrial dysfunction-induced β-cells apoptosis. Metabolic stress induced by antimycin or oligomycin was used to impair mitochondrial function in MIN6N8 cells, which are mouse pancreatic β-cells. Impaired mitochondria dysfunction increased ER stress proteins such as p-eIF2α, GRP78 and GRP 94, as well as ER stress-associated apoptotic factor,CHOP, and activated JNK. AMP-activated protein kinase (AMPK) was also activated under mitochondria dysfunction by metabolic stress. However, the inhibition of AMPK by treatment with compound C, inhibitor of AMPK, and overexpression of mutant dominant negative AMPK (AMPKK45R)blocked the induction of ER stress, which was consist-ent with the decreased β-cell apoptosis and increase of insulin content. Furthermore, mitochondrial dysfunction increased the expression of the inducible nitric oxide synthase (iNOS) gene and the production of nitric oxide (NO),but NO production was prevented by compound C and mutant dominant negative AMPK (AMPK-K45R). Moreover,treatment with 1400W, which is an inhibitor of iNOS, prevented ER stress and apoptosis induced by mitochondrial dysfunction. Treatment of MIN6N8 cells with lipid mixture,physiological conditions of impaired mitochondria function,activated AMPK, increased NO production and induced ER stress. Collectively, these data demonstrate that mitochondrial dysfunction activates AMPK, which induces ER stress via NO production, resulting in pancreatic β-cells apoptosis.

노르에피네프린 및 과활성산소-유도 스트레스 활성 단백질부활효소/c-Jun NH_(2)-말단 단백질부활효소(SAPK/JNK)의 반응성과 GaAIAs형 반도체 레이저의 노르에피네프린 억제

최유림,이숙희,최완석,김보경,김중환,김순희,민경옥 Yong-In University 2005 自然科學硏究所論文誌 Vol.10 No.1

The present study examined that In Vitro test is investigated in experimental animal and cell lines, such as measurement of phosphorylation, and that the antisympathetic effect was induced by specialized radiotherapy, such as LASER (light amplification by stimulated emission of radiution) of GaAIAs type, at meridian point (ST-36), in plasma analysis (norepinephrine) from rabbit. In western blotting analysis with anti-phosphorylated mitogen-activated protein kinase (MAPK) antibodies showed that the phosphorylation of SAPK-JNK (stress-activated protein kinase/c-Jun NH2-terminal protein kinase) was significantly increased, in a dose-and time-dependent manner, norepinephrine and H_(2)O_(2)-application state from the rat aortic muscle strips and smooth muscle cell lines (A7r5 cell), respectively. The plasma analysis showed that the concentration of norepinephrine was significantly increased in pushpin-stimulated group compared with control group. Furthermore, The GaAIAs type of semiconductor diode LASER, especially stimulated intensity of 140㎽, significantly decreased in plasma norepinephrine concentration from rabbit. Therefore, these results suggest that the application of norepinephrine and H_(2)O_(2) in aorta and A7r5 cell line is associated with changed phosphorylation of SAPK/JNK and that the diminished responsiveness (such as plasma concentrations of norepinephrine) to LASER in rabbit may be in part related by the increased of antisympathetic effects.

Expression profiling of the mitogen-activated protein kinase gene family reveals their diverse response pattern in two different salt-tolerant Glycyrrhiza species

Cao Aiping,Gao Ling,Wang Fei,Tong Xuechen,Xie Shuangquan,Chen Xifeng,Lu Tianxin,Shen Haitao,Liu Hailiang,Jin Xiang,Li Hongbin 한국유전학회 2022 Genes & Genomics Vol.44 No.7

Background: Mitogen-activated protein kinases (MPKs) play important role in response to environmental stress as crucial signal receptors or sensors. Our previous study indicated that salt stress acts as a positive factor to stimulate the production of pharmacodynamic metabolites in the medicinal plant Glycyrrhiza uralensis. Currently, little is known about the MPK gene family and their functions in the medicinal plant G. uralensis. Objective: Identification, comprehensive bioinformatic analysis, expression profiling, and response pattern under salt stress of the G. uralensis GuMPK gene family. Methods: Genome-wide investigation and expression profiling of the MPK gene family in G. uralensis, and their phylogenetic relationships, evolutionary characteristics, gene structure, motif distribution, promoter cis-acting element, and expression pattern under salt stress in two different salt-tolerant Glycyrrhiza species were performed. Results: A total of 20 G. uralensis GuMPK genes were identified and categorized into five groups, and had conserved gene structure and motif distribution. Expression profiling of GuMPK genes suggested their potentially diverse functions in plant growth and in response to phytohormones and environmental stress, particularly GuMPK1, 2, 5, and 10 as key components for G. uralensis in response to abiotic stress. Further expression analysis under NaCl treatment in two different salt-tolerant Glycyrrhiza species displayed the MPKs' different response patterns, emphasizing the role of MPK2, 5, 7, and 16 as potentially crucial genes for Glycyrrhiza to respond to salt stress. Conclusion: Our results provide a genome-wide identification and expression profiling of MPK gene family in G. uralensis, and establish the foundation for screening key responsive genes and understanding the potential function and regulatory mechanism of GuMPKs in salt responsiveness.

ER Stress Is Implicated in Mitochondrial Dysfunction-Induced Apoptosis of Pancreatic Beta Cells

Lee, June-Woo,Kim, Won-Ho,Yeo, Ji-Young,Jung, Myeong-Ho Korean Society for Molecular and Cellular Biology 2010 Molecules and cells Vol.30 No.6

Mitochondrial dysfunction induces apoptosis of pancreatic ${\beta}$-cells and leads to type 2 diabetes, but the mechanism involved in this process remains unclear. Chronic endoplasmic reticulum (ER) stress plays a role in the apoptosis of pancreatic ${\beta}$-cells; therefore, in current study, we investigated the implication of ER stress in mitochondrial dysfunction-induced ${\beta}$-cells apoptosis. Metabolic stress induced by antimycin or oligomycin was used to impair mitochondrial function in MIN6N8 cells, which are mouse pancreatic ${\beta}$-cells. Impaired mitochondria dysfunction increased ER stress proteins such as p-eIF2${\alpha}$, GRP78 and GRP 94, as well as ER stress-associated apoptotic factor, CHOP, and activated JNK. AMP-activated protein kinase (AMPK) was also activated under mitochondria dysfunction by metabolic stress. However, the inhibition of AMPK by treatment with compound C, inhibitor of AMPK, and overexpression of mutant dominant negative AMPK (AMPK-K45R) blocked the induction of ER stress, which was consistent with the decreased ${\beta}$-cell apoptosis and increase of insulin content. Furthermore, mitochondrial dysfunction increased the expression of the inducible nitric oxide synthase (iNOS) gene and the production of nitric oxide (NO), but NO production was prevented by compound C and mutant dominant negative AMPK (AMPK-K45R). Moreover, treatment with 1400W, which is an inhibitor of iNOS, prevented ER stress and apoptosis induced by mitochondrial dysfunction. Treatment of MIN6N8 cells with lipid mixture, physiological conditions of impaired mitochondria function, activated AMPK, increased NO production and induced ER stress. Collectively, these data demonstrate that mitochondrial dysfunction activates AMPK, which induces ER stress via NO production, resulting in pancreatic ${\beta}$-cells apoptosis.