Oxidative Stress by Mitochondrial Dysfunction Contributes to the Activation of Necroptosis in Acute Respiratory Distress Syndrome

( Su Hwan Lee ),( Ju Hye Shin ),( Ah Young Leem ),( Kyung Soo Chung ),( Jin Hwa Lee ),( Moo Suk Park ),( Jong-seok Moon ),( Young Sam Kim ) 대한결핵 및 호흡기학회 2021 대한결핵 및 호흡기학회 추계학술대회 초록집 Vol.129 No.-

Background Necroptosis is linked to cell death in Acute respiratory distress syndrome (ARDS). And oxidative stress has also been implicated in the pathogenesis of ARDS. Although the effect of necroptosis through oxidative stress is identified in the cellular system, the mechanisms for the activation of necroptosis in ARDS remain unclear. This study aims to confirm whether oxidative stress contributes to the receptor-interacting protein kinase 3 (RIPK3)-mediated necroptosis associated with ARDS. Methods The seven human lung tissues with ARDS were investigated for the involvement of RIPK3-mediated necroptosis and oxidative stress using immunoblot and immunostaining. We also investigated whether the oxidative stress due to mitochondrial dysfunction caused by oligomycin, an inhibitor of ATP synthase, might exacerbate the RIPK3-mediated necroptosis in the lipopolysaccharide (LPS)-induced acute lung injury (ALI) mouse model. Results We found that RIPK3-mediated necroptosis was significantly elevated in patients with ARDS compared to the non-ARDS subjects. Oxidative stress was increased in patients with ARDS due to the accumulation of 4-HNE, a marker for oxidative stress. Both RIPK3-mediated necroptosis and oxidative stress were elevated in the lung epithelial cells of the patients with ARDS. The oxidative stress significantly increased lung injury in the LPS-induced ALI mouse model compared to the ALI model without oligomycin treatment. Moreover, the oxidative stress significantly increased the levels of RIPK3 and the release of damageassociated molecular patterns by necroptosis in the lung during ALI compared to the group administered the dose without treatment of oligomycin. Conclusion The Results showed that RIPK3-mediated necroptosis and oxidative stress were increased in human ARDS, and the LPS lung injury mouse model with oxidative stress had increased RIPK3-mediated necroptosis, lung injury, and DAMPs release. Our Results suggest that oxidative stress due to mitochondrial dysfunction might contribute to a critical mechanism for RIPK3- mediated necroptosis-induced lung injury during ARDS.

Oxidative Stress and HSP70 Expression Upon Cerebral Isehemia-Reperfusion in Mongolian Gerbil

박영미,김철훈,도윤정,최은미,안영수,Park, Young-Mee,Kim, Chul-Hoon,Do, Yun-Jeong,Choi, Eun-Mi,Ahn, Young-Soo The Korean Society of Pharmacology 1996 대한약리학잡지 Vol.32 No.3

허혈/재관류 뇌손상에서 활성산소류의 역할이 중요시되고 있다. 본 연구에서는 모래쥐의 총경동맥을 묶었다 풀어줌으로써 실험적 허혈/재관류 손상을 유도하고 산화성 스트레스 발생 유무와 이러한 산화성 스트레스가 세포손상으로 연결되는지를 알아보고자 하였다. 해마는 뇌조직 중에서도 특히 산화성 스트레스에 취약한 부분이므로 해마에서 얻은 조직을 대뇌피질에서 얻은 조직과 비교분석하였다. 또한, 이들 부위에서 heat shock protein 70(HSP70)의 발현이 허혈/재관류 손상에 미치는 영향도 검색하고자 하였다. 허혈/재관류에 의한 산화성 스트레스의 지표로써 글루타치온 산화정도, GSSG/(GSH+2xGSSG)를 측정하였을 때 주로 해마에서 산화지표가 상승됨을 관찰하였다. 한편 산화성 스트레스가 세포손상으로 연결되는지를 알아보고자 지질과산화물을 측정하였다. 두 부위 모두에서 지질과산화물 형성의 증가가 있었으며 대뇌피질에서보다 해마에서 더 증가됨을 알 수 있었다. 지질과산화물 형성의 정도나 시간적 변화양상이 글루타치온 산화의 그것들과 유사하였다. 이러한 결과들은 허혈/재관류에 의해 산화성 스트레스가 형성되며 동시에 이러한 산화성 스트레스가 세포 손상을 초래함을 보여준다. 또한 산화성 스트레스 및 산화성 세포손상 정도가 대뇌피질보다는 해마에서 더 큰 것을 알 수 있었다. 그러나, 피질과 해마에서 HSP70의 기초발현(basal level) 정도는 차이가 없었다. 이는 해마의 취약성이 HSP70 발현 결핍에 기인하지 않았음을 나타낸다. 반면 허혈/재관류에 의한 HSP70의 발현유도는 해마조직에서 제대로 이주어지지 않았고 northern blot결과 이는 전사단계에서의 부친에 의한 것으로 나타났다. 이러한 결과들로 볼 때 허혈/재관류에 의한 뇌손상에서 HSP70 유도정도를 측정하는 것이 세포의 취약성을 예측할 수 있는 지표로 유용하게 사용될 수 있을 것으로 사료된다. A critical role of oxygen-derived free radicals has been implicated in ischemia/reperfusion (I/R)-induced brain damage. In this study, we have produced experimental I/R to the brains of Mongolian gerbil (Meriones unguiculatus) by a transient occlusion and release of the common carotid arteries. We have attempted to determine whether the oxidative stress is generated upon I/R and whether this oxidative stress is linked to the cell damage. Since hippocampus has been suggested as one of the most vulnerable regions of the brain to the oxidative stress, we analyzed samples from hippocampus in comparison with those from cortex. In addition, we have examined the expression of heat shock protein 70kD species (HSP70) in these regions in order to evaluate a possible role of this protein in I/R-induced brain damage. To determine whether the oxidative stress is produced upon I/R, we measured the glutathione oxidation, GSSG/ (GSH + 2xGSSG), as an index of oxidative stress. We found an increase of the glutathione oxidation primarily in hippocampus upon I/R. To determine whether this oxidative stress is linked to the cell damage, we measured the degree of lipid peroxidation upon I/R. We found an increase of lipid peroxidation in both regions. However, the magnitude of increases was greater in hippocampus than in cortex. In addition, we found that changes in both the magnitude and the temporal patterns of glutathione oxidation closely correlated with those of lipid peroxidation. Our study provides biochemical evidences that the oxidative stress is generated upon I/R and this oxidative stress is linked to the oxidative cell damage. Our study also provides evidences that the degree of oxidative stress as well as oxidative cell damage is greater in hippocampus than in cortex. We could not find difference in the basal level of HSP70 expression between hippocampus and cortex, indicating that the intrinsic vulnerability of hippocampus cannot be explained by the lower level of HSP70 expression. We did find, however, that the induction of HSP70 expression upon I/R was impaired in the hippocampus. This impairment appeared to be at the transcriptional level. These results suggest that the measurement of HSP70 induction may be employed as a useful predictor of differential cellular susceptibilities to the I/R-induced brain damage.

모래쥐에서 뇌의 허혈/재관류에 의한 산화성 스트레스 형성과 HSP70의 발현

박영미(Young-Mee Park),김철훈(Chul-Hoon Kim),도윤정(Yun-Jeong Do),최은미(Eun-Mi Choi),안영수(Young-Soo Ahn) 대한약리학회 1996 대한약리학잡지 Vol.32 No.3

허혈/재관류 뇌손상에서 활성산소류의 역할이 중요시되고 있다. 본 연구에서는 모래쥐의 총경동맥을 묶었다 풀어줌으로써 실험적 허혈/재관류 손상을 유도하고 산화성 스트레스 발생 유무와 이러한 산화성 스트레스가 세포손상으로 연결되는지를 알아보고자 하였다. 해마는 뇌조직 중에서도 특히 산화성 스트레스에 취약한 부분이므로 해마에서 얻은 조직을 대뇌피질에서 얻은 조직과 비교분석하였다. 또한, 이들 부위에서 heat shock protein 70(HSP70)의 발현이 허혈/재관류 손상에 미치는 영향도 검색하고자 하였다. 허혈/재관류에 의한 산화성 스트레스의 지표로써 글루타치온 산화정도, GSSG/(GSH+2xGSSG)를 측정하였을 때 주로 해마에서 산화지표가 상승됨을 관찰하였다. 한편 산화성 스트레스가 세포손상으로 연결되는지를 알아보고자 지질과산화물을 측정하였다. 두 부위 모두에서 지질과산화물 형성의 증가가 있었으며 대뇌피질에서보다 해마에서 더 증가됨을 알 수 있었다. 지질과산화물 형성의 정도나 시간적 변화양상이 글루타치온 산화의 그것들과 유사하였다. 이러한 결과들은 허혈/재관류에 의해 산화성 스트레스가 형성되며 동시에 이러한 산화성 스트레스가 세포 손상을 초래함을 보여준다. 또한 산화성 스트레스 및 산화성 세포손상 정도가 대뇌피질보다는 해마에서 더 큰 것을 알 수 있었다. 그러나, 피질과 해마에서 HSP70의 기초발현(basal level) 정도는 차이가 없었다. 이는 해마의 취약성이 HSP70 발현 결핍에 기인하지 않았음을 나타낸다. 반면 허혈/재관류에 의한 HSP70의 발현유도는 해마조직에서 제대로 이주어지지 않았고 northern blot결과 이는 전사단계에서의 부친에 의한 것으로 나타났다. 이러한 결과들로 볼 때 허혈/재관류에 의한 뇌손상에서 HSP70 유도정도를 측정하는 것이 세포의 취약성을 예측할 수 있는 지표로 유용하게 사용될 수 있을 것으로 사료된다. A critical role of oxygen-derived free radicals has been implicated in ischemia/reperfusion (I/R)-induced brain damage. In this study, we have produced experimental I/R to the brains of Mongolian gerbil (Meriones unguiculatus) by a transient occlusion and release of the common carotid arteries. We have attempted to determine whether the oxidative stress is generated upon I/R and whether this oxidative stress is linked to the cell damage. Since hippocampus has been suggested as one of the most vulnerable regions of the brain to the oxidative stress, we analyzed samples from hippocampus in comparison with those from cortex. In addition, we have examined the expression of heat shock protein 70kD species (HSP70) in these regions in order to evaluate a possible role of this protein in I/R-induced brain damage. To determine whether the oxidative stress is produced upon I/R, we measured the glutathione oxidation, GSSG/ (GSH + 2xGSSG), as an index of oxidative stress. We found an increase of the glutathione oxidation primarily in hippocampus upon I/R. To determine whether this oxidative stress is linked to the cell damage, we measured the degree of lipid peroxidation upon I/R. We found an increase of lipid peroxidation in both regions. However, the magnitude of increases was greater in hippocampus than in cortex. In addition, we found that changes in both the magnitude and the temporal patterns of glutathione oxidation closely correlated with those of lipid peroxidation. Our study provides biochemical evidences that the oxidative stress is generated upon I/R and this oxidative stress is linked to the oxidative cell damage. Our study also provides evidences that the degree of oxidative stress as well as oxidative cell damage is greater in hippocampus than in cortex. We could not find difference in the basal level of HSP70 expression between hippocampus and cortex, indicating that the intrinsic vulnerability of hippocampus cannot be explained by the lower level of HSP70 expression. We did find, however, that the induction of HSP70 expression upon I/R was impaired in the hippocampus. This impairment appeared to be at the transcriptional level. These results suggest that the measurement of HSP70 induction may be employed as a useful predictor of differential cellular susceptibilities to the I/R-induced brain damage.

Proteomic Analysis of Hippocampus in a Mouse Model of Depression Reveals Neuroprotective Function of Ubiquitin C-terminal Hydrolase L1 (UCH-L1) via Stress-induced Cysteine Oxidative Modifications

Choi, Jung-Eun,Lee, Jae-Jin,Kang, Wonmo,Kim, Hyun Jung,Cho, Jin-Hwan,Han, Pyung-Lim,Lee, Kong-Joo American Society for Biochemistry and Molecular Bi 2018 Molecular and Cellular Proteomics Vol.17 No.9

<P>Chronic physical restraint stress increases oxidative stress in the brain, and dysregulation of oxidative stress can be one of the causes of major depressive disorder. To understand the underlying mechanisms, we undertook a systematic proteomic analysis of hippocampus in a chronic restraint stress mouse model of depression. Combining two-dimensional gel electrophoresis (2D-PAGE) for protein separation with nanoUPLC-ESI-q-TOF tandem mass spectrometry, we identified sixty-three protein spots that changed in the hippocampus of mice subjected to chronic restraint stress. We identified and classified the proteins that changed after chronic stress, into three groups respectively functioning in neural plasticity, metabolic processes and protein aggregation. Of these, 5 proteins including ubiquitin C-terminal hydrolase L1 (UCH-L1), dihydropyrimidinase-related protein 2 (DPYL2), haloacid dehalogenase-like hydrolase domain-containing protein 2 (HDHD2), actin-related protein 2/3 complex subunit 5 (ARPC5) and peroxiredoxin-2 (PRDX2), showed pI shifts attributable to post-translational modifications. Further analysis indicated that UCH-L1 underwent differential oxidations of 2 cysteine residues following chronic stress. We investigated whether the oxidized form of UCH-L1 plays a role in stressed hippocampus, by comparing the effects of UCH-L1 and its Cys mutants on hippocampal cell line HT-22 in response to oxidative stress. This study demonstrated that UCH-L1 wild-type and cysteine to aspartic acid mutants, but not its cysteine to serine mutants, afforded neuroprotective effects against oxidative stress; there were no discernible differences between wild-type UCH-L1 and its mutants in the absence of oxidative stress. These findings suggest that cysteine oxidative modifications of UCH-L1 in the hippocampus play key roles in neuroprotection against oxidative stress caused in major depressive disorder.</P>

Negative Role of wblA in Response to Oxidative Stress in Streptomyces coelicolor

( Jin Su Kim ),( Han Na Lee ),( Pil Kim ),( Heung Shick Lee ),( Eung Soo Kim ) 한국미생물 · 생명공학회 2012 Journal of microbiology and biotechnology Vol.22 No.6

In this study, we analyzed the oxidative stress response of wblA (whiB-like gene A, SCO3579), which was previously shown to be a global antibiotic down-regulator in Streptomyces coelicolor. Ever since a WblA ortholog named WhcA in Corynebacterium glutamicum was found to play a negative role in the oxidative stress response, S. coelicolor wblA has been proposed to have a similar effect. A wblA-deletion mutant exhibited a less sensitive response to oxidative stress induced by diamide present in solid plate culture. Using real-time RT-PCR analysis, we also compared the transcription levels of oxidative stress-related genes, including sodF, sodF2, sodN, trxB, and trxB2, between S. coelicolor wild type and a wblA-deletion mutant in the presence or absence of oxidative stress. Target genes were expressed higher in the wblA-deletion mutant compared with wild type, both in the absence and presence of oxidative stress. Moreover, expression of these target genes in S. coelicolor wild type was stimulated only in the presence of oxidative stress, suggesting that WblA plays a negative role in the oxidative stress response of S. coelicolor, similar to that of C. glutamicum WhcA, through the transcriptional regulation of oxidative stress-related genes.

Expression of microRNAs associated with oxidative stress in the hippocampus of piglets

Binyu Yang,Hongyuan Mei,Fuyuan Zuo,Ling Gan 한국유전학회 2017 Genes & Genomics Vol.39 No.7

Oxidative stress is associated with human diseases and the developmental retardation of animals. The hippocampus is particularly vulnerable to oxidative stress. MicroRNAs (miRNAs), expressed largely in the mammalian brain, are emerging as robust players and have been implicated in many cellular processes. The present study investigated the sub-tissue specificity of miRNA expression in the dorsal hippocampus (DH) and ventral hippocampus (VH) and evaluated the effects of oxidative stress induced by iron dextran (FeDex) treatment on miRNA expression in the DH and VH of pigs using RNA-sequencing technology and bioinformatics, respectively. The results demonstrated that the injection of FeDex significantly increased the levels of several markers of oxidative stress in serum of Rongchang piglets, which indicated that oxidative stress was successfully induced. Sub-tissue specificity was displayed with 54 differentially expressed miRNAs between the VH and DH. The induced oxidative stress emphasized 59 and 46 differentially expressed miRNAs in the DH and VH, respectively. GO and KEGG pathway analyses revealed that the predicted targets of these differentially expressed miRNAs were involved in the pathways that regulate the expression of genes associated with nervous system development, immune response and oxidative stress, which not only revealed the ability of miRNAs to influence complex gene networks in the DH and VH but also further corroborated the successful induction of oxidative stress. Collectively, the results of this study provide a valuable basis for future studies aimed at contributions of miRNAs induced by oxidative stress in growth retardation and neurodegenerative diseases of animals and human.

정상인에서의 스트레스 반응 정도와 산화스트레스 사이의 상관관계 연구

변순임,김지영,조성훈,김종우,황의완,Byun, Soon-Im,Kim, Ji-Young,Cho, Seung-Hun,Kim, Jong-Woo,Hwang, Wei-Wan 대한예방한의학회 2009 대한예방한의학회지 Vol.13 No.2

Objective : This study was to examine a relationship among subjective psychological stress, physical stress response and oxidative stress. Methods : The subjects were consisted of 87 healthy volunteers. To assess subjective psychological stress, SRI(stress response inventory) was completed. To assess psychological and physical stress response, HRV(Heart Rate Variability) were tested. To assess oxidative stress, d-roms(Derivatives of reactive oxygen metabolites test) test was conducted. Subjects were divided into 3 groups depending on SRI(Stress Response Inventory) score, low stress response(LSR: lower than 30 percentile), Medium(MSR: 30-70 percentile) and high stress response(HSR: higher than 70 percentile). The Relationship between Stress Response and Oxidative Stress was estimated by correlation and One-way ANOVA analysis. Results : There were no significant differences of demographic data among 3 groups. There was a significant difference of oxidative stress among 3 groups. Conclusion : Our results suggest that there is a weak positive correlation between subjective psychological stress and oxidative stress. There is a significant difference of oxidative stress between HSR group and LSR group.

Pesticide metabolite and oxidative stress in male farmers exposed to pesticide

Kang Myoung Lee,Sang-Yoo Park,Kyungsuk Lee,Sung-Soo Oh,Sang Baek Ko 대한직업환경의학회 2017 대한직업환경의학회지 Vol.29 No.-

Background: The objective of this study was to measure malondialdehyde (MDA) and isoprostane which has been used as an index of lipid injury, 8-hydroxy-2′-deoxyguanosine (8-OHdG), which has been used as an index of DNA damage, and dialkyl-phosphate (DAP), which has been used to quantify pesticide exposure, and to investigate the relationship between pesticide exposure and oxidative stress. Methods: This study was a cross-sectional study that evaluated 84 male farmers exposure to pesticide. In this study, 8-OHdG, isoprostane, and MDA were measured as oxidative stress indices, and dialkyl-phosphate (dimethylphosphate(DMP), diethylphosphate(DEP), dimethylthiophosphate(DMTP), and diethylthiophosphate (DETP)) excreted in the urine was also measured to evaluate pesticide exposure. A linear regression analysis was performed to investigate the relationship between pesticide metabolites, and oxidative stress biomarkers. Results: A Correlation analysis was performed for pesticide exposure month (PEI), cumulative exposure index (CEI), and DAP as well as the concentration of the oxidative stress biomarkers. The PEM significantly and positively correlated to the levels of 8-OHdG, isoprostane, CEI, and DMP. CEI showed a correlation to 8-OHdG and PEM. DMP, DEP, and DETP showed a positive correlation to 8-OHdG, isoprostane, and MDA. A correlation analysis was adjusted some demographic characteristics, such as age, smoking, drinking, and exercise to determine the relationship between pesticide exposure and oxidative stress. The 8-OHdG, isoprostane, and MDA levels were significantly related to the DMP (ß = 0.320), DEP (ß = 0.390), and DETP (ß = 0.082); DMP (ß = 0.396), DEP (ß = 0.508), and DETP (ß = 0.504); and DMP (ß = 0.432), DEP (ß = 0.508), and DETP (ß = 0.329) levels, respectively. Conclusions: The concentration between oxidative stress biomarkers and the pesticide metabolite were a positive correlation. Indicators of oxidative stress was associated with a pesticide metabolite DMP, DEP, and DETP. Therefore, Pesticide exposure and oxidative stress were relevant.

Korean Red Ginseng and Korean black ginseng extracts, JP5 and BG1, prevent hepatic oxidative stress and infl ammation induced by environmental heat stress

송지현,김귀진,Sungwoo Chei,Young-Jin Seo,Kippeum Lee,Boo Yong Lee 고려인삼학회 2020 Journal of Ginseng Research Vol.44 No.2

Background: Continuous exposure to high temperatures can lead to heat stress. This stress responsealters the expression of multiple genes and can contribute to the onset of various diseases. In particular,heat stress induces oxidative stress by increasing the production of reactive oxygen species. The liver isan essential organ that plays a variety of roles, such as detoxification and protein synthesis. Therefore, itis important to protect the liver from oxidative stress caused by heat stress. Korean ginseng has a varietyof beneficial biological properties, and our previous studies showed that it provides an effective defenseagainst heat stress. Methods: We investigated the ability of Korean Red Ginseng and Korean black ginseng extracts (JP5 andBG1) to protect against heat stress using a rat model. We then confirmed the active ingredients andmechanism of action using a cell-based model. Results: Heat stress significantly increased gene and protein expression of oxidative stresserelatedfactors such as catalase and SOD2, but treatment with JP5 (Korean Red Ginseng extract) and BG1(Korean black ginseng extract) abolished this response in both liver tissue and HepG2 cells. In addition,JP5 and BG1 inhibited the expression of inflammatory proteins such as p-NF-kB and tumor necrosisfactor alpha-a. In particular, JP5 and BG1 decreased the expression of components of the NLRP3inflammasome, a key inflammatory signaling factor. Thus, JP5 and BG1 inhibited both oxidative stressand inflammation. Conclusions: JP5 and BG1 protect against oxidative stress and inflammation induced by heat stress andhelp maintain liver function by preventing liver damage.

Korean Red Ginseng and Korean black ginseng extracts, JP5 and BG1, prevent hepatic oxidative stress and inflammation induced by environmental heat stress

Song, Ji-Hyeon,Kim, Kui-Jin,Chei, Sungwoo,Seo, Young-Jin,Lee, Kippeum,Lee, Boo-Yong The Korean Society of Ginseng 2020 Journal of Ginseng Research Vol.44 No.2

Background: Continuous exposure to high temperatures can lead to heat stress. This stress response alters the expression of multiple genes and can contribute to the onset of various diseases. In particular, heat stress induces oxidative stress by increasing the production of reactive oxygen species. The liver is an essential organ that plays a variety of roles, such as detoxification and protein synthesis. Therefore, it is important to protect the liver from oxidative stress caused by heat stress. Korean ginseng has a variety of beneficial biological properties, and our previous studies showed that it provides an effective defense against heat stress. Methods: We investigated the ability of Korean Red Ginseng and Korean black ginseng extracts (JP5 and BG1) to protect against heat stress using a rat model. We then confirmed the active ingredients and mechanism of action using a cell-based model. Results: Heat stress significantly increased gene and protein expression of oxidative stress-related factors such as catalase and SOD2, but treatment with JP5 (Korean Red Ginseng extract) and BG1 (Korean black ginseng extract) abolished this response in both liver tissue and HepG2 cells. In addition, JP5 and BG1 inhibited the expression of inflammatory proteins such as p-NF-κB and tumor necrosis factor alpha-α. In particular, JP5 and BG1 decreased the expression of components of the NLRP3 inflammasome, a key inflammatory signaling factor. Thus, JP5 and BG1 inhibited both oxidative stress and inflammation. Conclusions: JP5 and BG1 protect against oxidative stress and inflammation induced by heat stress and help maintain liver function by preventing liver damage.