The inflammatory response of neutrophils in an in vitro model that approximates the postcardiac arrest state

Young-Duck Cho,Sung-Jun Park,Sung-Hyuk Choi,Young-Hoon Yoon,Jung-Youn Kim,Sung-Woo Lee,Chae-Seung Lim 대한외과학회 2017 Annals of Surgical Treatment and Research Vol.93 No.4

Purpose: Postcardiac arrest syndrome (PCAS) shares many features with sepsis including plasma cytokine elevation with dysregulation of cytokine production, and the presence of endotoxin in plasma. PCAS is closely related to ischemia-reperfusion injury. During ischemia-reperfusion injury, neutrophil, which is the first line of innate immunity, plays a major role. In this study, we investigated the inflammatory response of human neutrophils in an in vitro model which we simulated with hypoxia-normoxia and hypoxia-hyperoxia environments. Methods: After separation of neutrophils from the whole blood, they were divided into 3 experimental groups: normoxianormoxia, hypoxia-normoxia, and hypoxia-hyperoxia groups. The production of H2O2, the expression of Toll-like receptor 4 (TLR4) receptor, and the extent of apoptosis of the neutrophils were checked. Results: The in vitro hypoxia-normoxia and -hyperoxia models, which simulated the PCAS, showed initiation of the neutrophils’ inflammatory reaction by hypoxia insult. Lipopolysaccharide amplifies such inflammation; therefore, prevention of secondary infection may be critical in postresuscitation patients. Temporary hyperoxia following hypoxic insult showed no difference in inflammatory reaction compared with hypoxia-normoxia. Rather, temporary hyperoxia may suppress or minimize inflammation by attenuation of TLR4 receptor. Conclusion: It is well known that continuous hyperoxygenation after successful cardiac arrest harms patients, but temporary hyperoxygenation with 100% O2 in a clinical situation may be helpful.

The inflammatory response of neutrophils in an <i>in vitro</i> model that approximates the postcardiac arrest state

Cho, Young-Duck,Park, Sung-Jun,Choi, Sung-Hyuk,Yoon, Young-Hoon,Kim, Jung-Youn,Lee, Sung-Woo,Lim, Chae-Seung The Korean Surgical Society 2017 Annals of Surgical Treatment and Research(ASRT) Vol.93 No.4

<P><B>Purpose</B></P><P>Postcardiac arrest syndrome (PCAS) shares many features with sepsis including plasma cytokine elevation with dysregulation of cytokine production, and the presence of endotoxin in plasma. PCAS is closely related to ischemia-reperfusion injury. During ischemia-reperfusion injury, neutrophil, which is the first line of innate immunity, plays a major role. In this study, we investigated the inflammatory response of human neutrophils in an <I>in vitro</I> model which we simulated with hypoxia-normoxia and hypoxia-hyperoxia environments.</P><P><B>Methods</B></P><P>After separation of neutrophils from the whole blood, they were divided into 3 experimental groups: normoxia-normoxia, hypoxia-normoxia, and hypoxia-hyperoxia groups. The production of H<SUB>2</SUB>O<SUB>2</SUB>, the expression of Toll-like receptor 4 (TLR<SUB>4</SUB>) receptor, and the extent of apoptosis of the neutrophils were checked.</P><P><B>Results</B></P><P>The <I>in vitro</I> hypoxia-normoxia and -hyperoxia models, which simulated the PCAS, showed initiation of the neutrophils' inflammatory reaction by hypoxia insult. Lipopolysaccharide amplifies such inflammation; therefore, prevention of secondary infection may be critical in postresuscitation patients. Temporary hyperoxia following hypoxic insult showed no difference in inflammatory reaction compared with hypoxia-normoxia. Rather, temporary hyperoxia may suppress or minimize inflammation by attenuation of TLR<SUB>4</SUB> receptor.</P><P><B>Conclusion</B></P><P>It is well known that continuous hyperoxygenation after successful cardiac arrest harms patients, but temporary hyperoxygenation with 100% O<SUB>2</SUB> in a clinical situation may be helpful.</P>

The inflammatory response of neutrophils in an in vitro model that approximates the postcardiac arrest state

조영덕,박성준,최성혁,윤영훈,김정윤,이성우,임채승 대한외과학회 2017 Annals of Surgical Treatment and Research(ASRT) Vol.93 No.4

Purpose: Postcardiac arrest syndrome (PCAS) shares many features with sepsis including plasma cytokine elevation with dysregulation of cytokine production, and the presence of endotoxin in plasma. PCAS is closely related to ischemia-reperfusion injury. During ischemia-reperfusion injury, neutrophil, which is the first line of innate immunity, plays a major role. In this study, we investigated the inflammatory response of human neutrophils in an in vitro model which we simulated with hypoxia-normoxia and hypoxia-hyperoxia environments. Methods: After separation of neutrophils from the whole blood, they were divided into 3 experimental groups: normoxianormoxia, hypoxia-normoxia, and hypoxia-hyperoxia groups. The production of H2O2, the expression of Toll-like receptor 4 (TLR4) receptor, and the extent of apoptosis of the neutrophils were checked. Results: The in vitro hypoxia-normoxia and -hyperoxia models, which simulated the PCAS, showed initiation of the neutrophils’ inflammatory reaction by hypoxia insult. Lipopolysaccharide amplifies such inflammation; therefore, prevention of secondary infection may be critical in postresuscitation patients. Temporary hyperoxia following hypoxic insult showed no difference in inflammatory reaction compared with hypoxia-normoxia. Rather, temporary hyperoxia may suppress or minimize inflammation by attenuation of TLR4 receptor. Conclusion: It is well known that continuous hyperoxygenation after successful cardiac arrest harms patients, but temporary hyperoxygenation with 100% O2 in a clinical situation may be helpful

저산소/재관류로부터 청폐사간탕의 PC12 세포 보호 효과

소윤조 한국생약학회 2005 생약학회지 Vol.36 No.2

This research was performed to investigate the protective effect of Chungpesagan-tang (CST) from hypoxia /reperfusion induced-PC12 cell damage. To elucidate the mechanism of the protective effect of CST, cell viability, changes in activities of superoxide dismutase, glutathione peroxidase, catalase, caspase 3 and the production of malondialdehyde were observed after treating PC12 cells with CST which was metabolized by rat liver homogenate. Pretreatment of CST with liver homogenate appeared to increase its protective effect against hypoxia/reperfusion insult. The result showed that CST exhibited the highest protective effect against hypoxia/reperfusion at the dose of 1 µg/ml in PC12 cells, probably by recovering the redox enzyme activities and MDA to control level.

간효소에 의해 대사된 양격산화탕의 저산소/재관류로부터 PCl2 세포 보호효과

소윤조(Yunjo Soh) 大韓藥學會 2005 약학회지 Vol.49 No.1

The protective effect of Yangguksanwha-tang (YST) against hypoxia-reperfusion insult was investigated in PC12 cells. To elucidate the mechanism of the protective effect of YST, cell viability the changes in activities of superoxide dismutase, glutathione peroxidase, catalase, caspase 3 and the production of malondialdehyde were observed after treating PC12 cells with YST which was metabolized by rat liver homogenate. Pretreatment of YST with liver homogenate appeared to increase its protective effect against hypoxia-reperfusion insult. The result showed that YST had the highest protective effect against hypoxia/reperfusion at the dose of 2㎍/ml in PC12 cells, probably by recovering the redox enzyme activities and MDA to control level.

뇌의 저산소증-허혈과 재산소화-재관류가 유발된 신생자돈에서 7-Nitroindazole이 뇌세포막 기능과 에너지 대사에 미치는 영향에 대한 연구

최창원,황종희,장윤실,박원순 대한소아과학회 2004 小兒科 Vol.47 No.2

Is Biochemical Follow Up Possible in Peripheral Arterial Disease Treatment: Hypoxia Inducible Factor-1 Alpha?

Cihan Yücel,Mete Gürsoy,Serkan Ketenciler,Aslıhan Tenekeciğil,Feryaz Kızıltan,Nihan Kayalar 대한혈관외과학회 2021 Vascular Specialist International Vol.37 No.3

Purpose: The hypoxia inducible factor (HIF)-1 is a dimeric protein complex that plays an integral role in the body’s response to hypoxia. This study aimed to analyze the regulation of HIF-1α following vascular and/or endovascular surgery in peripheral arterial disease (PAD) patients. Materials and Methods: A total of 40 patients with PAD (≥Rutherford category 3) were included in this prospective study. The mean age was 61.9±9.2 years. Open surgery was performed in 16 patients, and endovascular intervention was performed in 34 patients. At preoperative (T1), postoperative day 1 (T2), and month 3 (T3), the serum HIF-1α levels were checked using the ELISA technique. Results: At T3, the ankle-brachial index was significantly higher than the preoperative value (P<0.001). Serum HIF-1α levels at T1, T2, and T3 were 2.0±1.7 ng/mL, 1.9±1.7 ng/mL, and 1.6±1.4 ng/mL, respectively. Serum HIF-1α levels between T1 and T3 and between T2 and T3 were significantly different (P<0.05). The preoperative HIF-1α levels were lowest in iliac lesions compared to femoropopliteal or tibial lesions. Conclusion: The HIF-1α levels were decreased in all patients on postoperative days, T2 and T3, compared with the preoperative values. Our results indicated that HIF-1α may be a surrogate marker after revascularization in patients with PAD. Further studies are needed to analyze the sensitivity, specificity, and cut-off values of HIF-1α in patients with PAD.

CircZNF609 Aggravated Myocardial Ischemia Reperfusion Injury via Mediation of miR-214-3p/PTGS2 Axis

Wen-Qiang Tang,Feng-Rui Yang,Ke-Min Chen,Huan Yang,Yu Liu,Bo Dou 대한심장학회 2022 Korean Circulation Journal Vol.52 No.9

Background and Objectives: Circular RNAs were known to play vital role in myocardial ischemia reperfusion injury (MIRI), while the role of CircZNF609 in MIRI remains unclear. This study was aimed to investigate the function of CircZNF609 in MIRI. Methods: Hypoxia/reoxygenation (H/R) model was established to mimic MIRI in vitro. Quantitative polymerase chain reaction was performed to evaluate gene transcripts. Cellular localization of CircZNF609 and miR-214-3p were visualized by fluorescence in situ hybridization. Cell proliferation was determined by CCK-8. TUNEL assay and flow cytometry were applied to detect apoptosis. Lactate dehydrogenase was determined by commercial kit. ROS was detected by DCFH-DA probe. Direct interaction of indicated molecules was determined by RIP and dual luciferase assays. Western blot was used to quantify protein levels. In vivo model was established to further test the function of CircZNF609 in MIRI. Results: CircZNF609 was upregulated in H/R model. Inhibition of CircZNF609 alleviated H/R induced apoptosis, ROS generation, restored cell proliferation in cardiomyocytes and human umbilical vein endothelial cells. Mechanically, CircZNF609 directly sponged miR-214-3p to release PTGS2 expression. Functional rescue experiments showed that miR-214-3p/PTGS2 axis was involved in the function of circZNG609 in H/R model. Furthermore, data in mouse model revealed that knockdown of CircZNF609 significantly reduced the area of myocardial infarction and decreased myocardial cell apoptosis. Conclusions: CircZNF609 aggravated the progression of MIRI via targeting miR-214-3p/PTGS2 axis, which suggested CircZNF609 might act as a vital modulator in MIRI.

EGb 761 Protects Cardiac Microvascular Endothelial Cells against Hypoxia/Reoxygenation Injury and Exerts Inhibitory Effect on the ATM Pathway

( Chao Zhang ),( Deng-feng Wang ),( Zhuang Zhang ),( Dong Han ),( Kan Yang ) 한국미생물 · 생명공학회 2017 Journal of microbiology and biotechnology Vol.27 No.3

Ginkgo biloba extract (EGb 761) has been widely used clinically to reduce myocardial ischemia reperfusion injury (MIRI). Microvascular endothelial cells (MVECs) may be a proper cellular model in vitro for the effect and mechanism study against MIRI. However, the protective effect of EGb 761 on MVECs resisting hypoxia/reoxygenation (H/R) injury is little reported. In this study, H/R-injured MVECs were treated with EGb 761, and then the cell viability, apoptosis, ROS production, SOD activity, caspase-3 activity, and protein level of ATM, γ-H2AX, p53, and Bax were measured. ATM siRNA was transfected to study the changes of protein in the ATM pathway. EGb 761 presented protective effect on H/R-injured MVECs, with decreasing cell death, apoptosis, and ROS, and elevated SOD activity. Next, EGb 761 could inhibit H/R-induced ATM, γ-H2AX, p53, and Bax in a dose-dependent manner. Moreover, ATM siRNA also could inhibit H/R-induced ATM, γ-H2AX, p53, and Bax. Overall, these findings verify that EGb 761 protects cardiac MVECs from H/R injury, and for the first time, illustrate the influence on the ATM pathway and apoptosis by EGb 761 via dampening ROS.

Augmenter of Liver Regeneration Alleviates Renal Hypoxia-Reoxygenation Injury by Regulating Mitochondrial Dynamics in Renal Tubular Epithelial Cells

Long, Rui-ting,Peng, Jun-bo,Huang, Li-li,Jiang, Gui-ping,Liao, Yue-juan,Sun, Hang,Hu, Yu-dong,Liao, Xiao-hui Korean Society for Molecular and Cellular Biology 2019 Molecules and cells Vol.42 No.12

Mitochondria are highly dynamic organelles that constantly undergo fission and fusion processes that closely related to their function. Disruption of mitochondrial dynamics has been demonstrated in acute kidney injury (AKI), which could eventually result in cell injury and death. Previously, we reported that augmenter of liver regeneration (ALR) alleviates renal tubular epithelial cell injury. Here, we gained further insights into whether the renoprotective roles of ALR are associated with mitochondrial dynamics. Changes in mitochondrial dynamics were examined in experimental models of renal ischemia-reperfusion (IR). In a model of hypoxia-reoxygenation (HR) injury in vitro, dynamin-related protein 1 (Drp1) and mitochondrial fission process protein 1 (MTFP1), two key proteins of mitochondrial fission, were downregulated in the Lv-ALR + HR group. ALR overexpression additionally had an impact on phosphorylation of Drp1 Ser637 during AKI. The inner membrane fusion protein, Optic Atrophy 1 (OPA1), was significantly increased whereas levels of outer membrane fusion proteins Mitofusin-1 and -2 (Mfn1, Mfn2) were not affected in the Lv-ALR + HR group, compared with the control group. Furthermore, the mTOR/4E-BP1 signaling pathway was highly activated in the Lv-ALR + HR group. ALR overexpression led to suppression of HR-induced apoptosis. Our collective findings indicate that ALR gene transfection alleviates mitochondrial injury, possibly through inhibiting fission and promoting fusion of the mitochondrial inner membrane, both of which contribute to reduction of HK-2 cell apoptosis. Additionally, fission processes are potentially mediated by promoting tubular cell survival through activating the mTOR/4E-BP1 signaling pathway.