내독소에 의한 말초혈액 단핵구의 IL-8 및 IL-$1{\beta}$ 유전자 발현에서 산소기 역할에 관한 연구

강민종,김재열,박재석,이승준,유철규,김영환,한성구,심영수,Kang, Min-Jong,Kim, Jae-Yeol,Park, Jae-Seok,Lee, Seung-Joon,Yoo, Chul-Gyu,Kim, Young-Whan,Han, Sung-Koo,Shim, Young-Soo 대한결핵및호흡기학회 1995 Tuberculosis and Respiratory Diseases Vol.42 No.6

연구배경: 산소기의 작용은 과거에는 세포독성이 주로 알려져 있었던 반면, 최근 들어 산소기의 세포내 신호전달체계에서의 역할에 많은 사람의 관심이 모이고 있다. 여러 cytokine의 전사인자(transcription factor)로 작용하는 $NF{\kappa}B$는 기저상태에서는 세포질에 존재하는데 $I{\kappa}B$와 결합되어 핵내로의 이동이 억제되고 있다. 여러 연구에 의해 $NF{\kappa}B$의 $I{\kappa}B$로부터의 분리는 외부자극에 의해 생성된 산소기에 의한 것으로 알려졌는데, 이렇게 하여 분리된 $NF{\kappa}B$가 핵내로 이동하면 핵내에서 전사인자로 작용하여 여러 유전자의 전사를 증가시키는 것이 보고되었다. IL-8 유전자는 5'flanking promotor region에 $NF{\kappa}B$-like motif가 있어 핵내 $NF{\kappa}B$ activity의 증가로 IL-8 유전자의 전사가 증가되는 것으로 알려졌고, 또한 내독소는 핵내의 $NF{\kappa}B$ activity의 증가와 함께 호중구에서의 산소기의 분비를 가져온다. 이러한 사실로부터 내독소에 의한 IL-8 유전자의 발현은 세포내에서 생성된 산소기에 의해 $NF{\kappa}B$가 $I{\kappa}B$로부터 분리되어 핵내로 이동하고 이로 인해 IL-8 유전자의 전사가 증가되는 가설을 생각할 수 있다. 저자들은 이러한 가설 검정의 첫번째 단계로써 체내 염증반응에서 중요한 역할을 하는 말초혈액 단핵구의 IL-8 및 IL-$1{\beta}$ 유전자 발현에 세포내의 산소기가 관여하는지의 여부를 평가하고자 본 연구를 시행하였다. 방법: Ficoll-Hypaque density gradient 법과 plastic 부착법을 이용하여 말초혈액 단핵구를 분리하였다. 외부에서 투여한 산소기의 농도에 따른 IL-8 및 IL-$1{\beta}$ mRNA 발현의 유무를 관찰하기 위하여 $H_2O_2$를 0, 10, 100, $300{\mu}M/L$, 1mM/L의 농도로 투여하고 6시간이 경과한후 IL-8 및 IL-$1{\beta}$에 대한 Northern blot analysis를 시행하였다. 시간에 따른 IL-8 및 IL-$1{\beta}$ mRNA 변화를 관찰하고자 $H_2O_2$를 $100{\mu}M/L$의 농도로 투여하고 0, 2, 4, 6, 8, 24시간이 경과한 후 Northern blot analysis를 시행하였다. 항산화제가 내독소에 의한 IL-8과 IL-$1{\beta}$ mRNA 발현에 미치는 영향을 평가하기 위하여 TMTU(10 mM/L) 1시간; PDTC($100{\mu}M/L$) 1시간, NAC(10 mM/L) 2.5시간, ME(10mM/(L) 2.5시간, Desferrioxamine(100mM/L) 15시간 동안 전처치 한 디음 내독소를 투여허여 4시간이 경과한 후 IL-8 및 IL-$1{\beta}$ mRNA에 대한 Northern blot analysis를 시행하였다. 결과: $H_2O_2$농도 및 시간에 따른 말초혈액 단핵구에서의 IL-8 및 IL-$1{\beta}$ mRNA의 발현에는 유의한 차이가 관찰되지 않았지만 항산화제로 전처치하였을 때 내독소에 의한 말초혈액 단핵구에서의 IL-8 및 IL-$1{\beta}$ mRNA의 발현이 억제되었고 그 억제정도는 TMTU에서 가장 현저하였다. 결론: 이상의 결과에서 말초혈액 단핵구에서의 IL-8 및 IL-$1{\beta}$ mRNA 발현에 $H_2O_2$가 아닌 다른 산소기가 일부 관여할 것으로 생각된다. Background: Oxygen free radicals have generally been considered as cytotoxic agents. On the other hand, recent results suggest that small nontoxic amounts of these radicals may act a role in intracellular signal transduction pathway and many efforts to reveal the role of these radicals as secondary messengers have been made. It is evident that the oxygen radicals are released by various cell types in response to extracellular stimuli including LPS, TNF, IL-1 and phorbol esters, all of which translocate the transcription factor $NF{\kappa}B$ from cytoplasm to nucleus by releasing an inhibitory protein subunit, $I{\kappa}B$. Activation of $NF{\kappa}B$ is mimicked by exposure to mild oxidant stress, and inhibited by agents that remove oxygen radicals. It means the cytoplasmic form of the inducible tanscription factor $NF{\kappa}B$ might provide a physiologically important target for oxygen radicals. At the same time, it is well known that LPS induces the release of oxygen radicals in neutrophil with the activation of $NF{\kappa}B$. From above facts, we can assume the expression of IL-8 and IL-$1{\beta}$ gene by LPS stimulation may occur through the activation of $NF{\kappa}B$, which is mediated through the release of $I{\kappa}B$ by increasing amounts of oxygen radicals. But definitive evidence is lacking about the role of oxygen free radicals in the expression of IL-8 and IL-$1{\beta}$ gene in mononuclear phagocytic cells. We conducted a study to determine whether oxygen radicals act a role in the expression of IL-8 and IL-$1{\beta}$ gene in mononuclear phagocytic cells. Method: Human peripheral blood monocytes were isolated from healthy volunteers. Time and dose relationship of $H_2O_2$-induced IL-8 and IL-$1{\beta}$ mRNA expression was observed by Northern blot analysis. To evaluate the role of oxygen radicals in the expression of IL-8 and IL-$1{\beta}$ mRNA by LPS stimulation, pretreatment of various antioxiants including PDTC, TMTU, NAC, ME, Desferrioxamine were done and Northern blot analysis for IL-8 and IL-$1{\beta}$ mRNA was performed. Results: In PBMC, dose and time dependent expression of IL-8 and IL-$1{\beta}$ mRNA by exogenous $H_2O_2$ was not observed. But various antioxidants suppressed the expression of LPS-induced IL-8 and IL-$1{\beta}$ mRNA expression of PBMC and the suppressive activity was most prominant when the pretreatment was done with TMTU. Conclusion: Oxygen free radical may have some role in the expression of IL-8 and IL-$1{\beta}$ mRNA of PBMC but that radical might not be $H_2O_2$.

비용에서 스테로이드 투여 후 활성산소의 생성과 항산화효소 활성도의 변화

조진학,김현웅,김희동,오원희,정인교,정근,임현준 대한이비인후과학회 2000 대한이비인후과학회지 두경부외과학 Vol.43 No.1

활성산소(活性酸素)에 의한 산화스트레스환자의 사상체질별(四象體質別) 특성(特性)에 관한 연구(硏究)

한경수,안택원,배나영,Han, Kyoung-Su,Ahn, Taek-Won,Bae, Na-Young 사상체질의학회 2006 사상체질의학회지 Vol.18 No.3

1. Objectives The purpose of this research is to find out the Constitutional difference of Oxidative stress by Free oxygen radicals. 2. Methods We enrolled 55 patients who visited our hospital for medical examination from February 1, 2006 to May 31, 2006. This research investigated the Constitutional difference of Oxidative stress by Free oxygen radicals. 3. Results and Conclusions l) The Sasang Constitutional distribution of patients have oxidatative stress by Free oxygen radicals was Taeunmin 30%(8persons of total 27persons), Soeumin 56%(9persons of total 16persons), Soyangin 33%(4persons of total 12persons). The rate of patients have oxidatate stress was high in Soeumin. The Sasang Constitution had significance relation with oxidatative stress by Free oxygen radicals. 2) Triglyceride on the average of Taeumin patients have oxidatative stress by Free oxygen radicals is ststistical significantly higher than Triglyceride on the average of controlled normal persons in Taeumin(p-value=0.010) and Soyangin(p-value=0.015). Blood uric acid level on the average of patients have oxidatate stress by Free oxygen radicals is ststistical significantly lower than Blood uric acid level on the average of controlled normal persons in Taeumin(p-value=0.004) and Soyangin(p-value=0.037). This research has shown that there is a statistical significance between the Triglyceride level, Blood uric acid level and oxidatate stress by Free oxygen radicals.

SOD 및 DMTU 가 자유산소기의 작용으로 배양중인 흰쥐섬유모세포에서 나타나는 손상에 미치는 영향

안동춘,강현국,백두진,정호삼 한양대학교 의과대학 1999 한양의대 학술지 Vol.19 No.1

This study performed to examine the effects of SOD and DMTU on the cytotoxicity of the cultured fibroblasts induced by oxygen free radicals. The rat fibroblasts was obtained from the subcutaneous tissue and the culture cells were exposed to hypoxanthine(HX) (1mM/ml) and xanthine oxidase(XO) at concentrations of 1, 5, 10 and 20 mU/ml, respectively. Then the cells exposed oxygen free radicals were treated by superoxide dismutase(SOD), superoxide scavenger, and 1, 3-dimethyl-2-thiourea(DMTU), hydroxyl radical scavenger, respectively. After 4 hours, the cells were observed under the inverted microscope and the cell viability was measured by MTT assay. The results as follows; 1. The morphologic changes derived by oxygen free radicals was observed at the 1mM HX plus 10mU and 20mU XO. 2. The survival rates of the cells were 88.0±8.5%, 24.0±4.4%, 12.0±1.0%, and 6.3±0.6% at the 1mM HX plus XO at concentration of 1, 5, 10, 20mU, respectively. 3. The viability of cells treated with SOD, one of the superoxide anion scavenger, was 90.7±16.3%, 66.0±9.8%, 32.7±6.7%, 16.3±2.5% at concentration of 1, 5, 10, 20mU XO, respectively. 4. There were no difference of cell viability between the DMTU treated cells and untreated well cells. These results suggest that the oxygen free radical cytotoxicity generated by HX and XO occur in the rat fibroblasts, and SOD can prevente its effects in vitro.

대장암의 생성과정에서의 활성산소의 영향

이명수,박영진,장석효 인제대학교 백병원 2002 仁濟醫學 Vol.23 No.2

Objective: The oxygen free radical causes DNA damage and consequent cancer development. However, the implication of oxygen free radical on colon carcinogenesis was not clearly demonstrated. This study was aimed to test the involvement of oxygen free radical on colon carcinogenesis. Methods and Materials: The activities of antioxidant enzymes (catalase and SOD), the extent of oxidative cellular damage (the content of 8ohGua) and the DNA repairing ability (oh8Gua glycosylase) was measured in 20 paired normal and tumorous colon specimens. Results: The antioxidant activity was decreased (p<0.01 in both the catalase and SOD) in tumor tissue than in normal. Contrary, the extent of oxidative DNA damage (content of 8ohGua) was increased by 1.6 times in tumor tissue (p<0.01). The overall activity of 8ohGua glycosylase was higher in tumors than in normal tissues (4.5 and 2.5 pmol oh8Gua released/2hr respectively, p=0.01). Conclusion: Although our sample size is limited, these results indicate that the greater oxidative DNA damage in cancer tissue as a possible result of the impaired antioxidant activity implies an important role of oxygen free radical in colon carcinogenesis. The DNA repairing activity enhance in colon cancer in response to the oxidative DNA damage, however, it is not sufficient enough to fully compensate the adverse effect of oxygen free radical.

Effects of boron/nitrogen/phosphorus doping on the scavenging action of armchair single‑walled carbon nanotubes (armchair‑SWCNT) for OH radicals: a DFT study

Meenakshi Malakar,Pradeep Kumar Shukla 한국탄소학회 2023 Carbon Letters Vol.33 No.1

Hydroxyl radical (OH radical) is the most harmful free radical amongst the Reactive Oxygen Species (ROS) responsible for numerous diseases of DNA damage like mutagenesis, carcinogenesis and ageing. Therefore, it is important to find a suitable scavenger for OH radical. In the present contribution, we aim to investigate the ability of pristine armchair-SWCNT and B/N/P-doped armchair-SWCNT to scavenge OH radicals using DFT calculations. The calculations reveal that the B/Pdoped armchair-SWCNTs can act as a better scavenger for OH radical compared to pristine armchair-SWCNT but N-doped armchair-SWCNT does not act as a better scavenger for OH radical compared to pristine armchair-SWCNT. Furthermore, the developed scavenger is examined in terms of large-scale availability, biocompatibility, conductivity, stability and reactivity. For both in vivo and in vitro studies, the work is found to useful for enhancing SWCNT as a free radical scavenger.

Myoglobin에 의한 급성 신부전에서 Oxygen Free Radical의 역할

김주섭 ( Joo Seop Kim ),정대윤 ( Dae Yune Jung ),김장만 ( Jang Man Kim ),조홍래 ( Hong Rae Cho ),배수동 ( Soo Tong Pai ),강구 ( Gu Kang ),신형식 ( Hyung Sik Shin ) 대한외상학회 1991 大韓外傷學會誌 Vol.4 No.2

Highly reactive oxygen metabolites have been known to have a role in several models of acute renal failure. Especially hydroxyl radicals have been proved to be important mediators by some previous authors. This study was aimed to determine the role of superoxide radical as well as hydroxyl radical and to compare the protective effects of both scavengers in myoglobinuric renal failure. Acute renal failure was induced by intramuscular injection of hypertonic glycerol (6mg/kg, half-dose in each hindlimb muscles) in rats. Rats treated with glycerol and hydroxyl radical scavenger, dimethylsulfoxide (DMSO), showed lower serum blood urea nitrogen (BUN) and creatinine level than rats with glycerol alone. And also significant protection was noticed histologically. Superoxide radical scavenger, superoxide dismutase (SOD), had a similar protective effects. In contrast, allopurinol, an inhibitor of xanthine oxidase, was not protective at all. The present study may suggest that both superoxide and hydoxyl radical playa role of tissue injury in myoglobin-induced renal failure. Beside xanthine oxidase, other souces of oxygen free radical may exist.

청아환(靑娥丸)에 의한 활성(活性) 산소류(酸素類)의 소거(消去) 작용(作用)과 항산화(抗酸化) 효소계(酵素系)의 활성(活性) 증가(增加) 효과(效果)에 대(對)한 연구(硏究)

정지천,Jeong, Ji-Cheon 대한한의학회 1997 대한한의학회지 Vol.18 No.2

This study was undertaken to examine the effect of Cheongahwan(CAH), being known to reinforce Kidney-yang, on the activities of endogenous antioxidant enzymes and the production of oxygen free radicals in the kidney tissues. Alterations in enzyme activities were observed after in vivo treatment in rats. CAH caused a significant increase in the activities of superoxide dismutase (SOD), glutathione peroxidase and glutathione S-transferase. But catalase activity was not significantly altered by CAH. Treatment in vitro of CAH decreased the production of oxygen free radicals in a dose-dependent fashion. These results suggest that CAH stimulate the activities of antioxidant enzymes and inhibit directly the production of oxygen free radicals. These effects of CAH may contribute to prevent the oxygen free radical-induced impairment of cell function.

일산화탄소 중독시 고압산소투여가 8-hydroxydeoxyguanosine 생성에 미치는 영향

김헌,조수헌,정명희,Kim, Heon,Cho, Soo-Hun,Chung, Myung-Hee 대한예방의학회 1994 Journal of Preventive Medicine and Public Health Vol.27 No.1

Hyperbaric oxygen (HBO) therapy for carbon monoxide (CO) poisoning eventually inducing the hypoxia-reoxygenation condition, may produce oxygen free radicals, which forms 8-hydroxydeoxyguanosine (8-OH-dG) by attacking C-8 position of deoxyguanosine (dG) in DNA. Effects of oxygen partial pressure or duration of HBO therapy with or without CO poisoning on the tissue 8-OH-dG formation were investigated. Male Sprague-Dawley rats were grouped and exposed to air (control group), 4000 ppm of CO for 10 to 30 minutes (CO only group), air for 30 minutes after 30 minute exposure to 4000 ppm of CO(CO-air exposure group), HBO after 30 minute exposure to 4000 ppm of CO(CO-HBO group), or HBO therapy fo. $10{\sim}120$ minutes(HBO only group). The 8-OH-4G concentrations in the brain and the lung tissues were measured with high performance liquid chromatography and electrochemical detector (ECD). Average concentrations of the 8-OH-dG of each group were statistically compared. In the brain tissues, 8-OH-dG concentrations of the CO only group, the CO-air exposure group, and the CO-HBO group did not significantly differ from those of the control group. Similar insignificance was also found between the CO-HBO group and the HBO only groups. No appreciable dose-response relationship was observed between the 8-OH-dG concentration and the oxygen partial pressure or the duration of HBO. However, the 8-OH-dG concentrations of the 30 minute CO only group were higher than those of the CO-air exposure group (p-value<0.05). In the lung tissues, there were no significant differences between the 8-OH-dG concentrations of the control group and those of the CO only group, the CO-air exposure group, and the CO-HBO group. However, mean 8-OH-dG concentration of the CO-air exposure group was significantly higher than that of the CO only group under the same CO exposure condition(p-value<0.05). With the duration of CO exposure, the 8-OH-dG concentrations of the lung tissues decreased significantly (p-value<0.05). The concentrations of 8-OH-dG in the lung tissues proportionally increased with the duration of HBO, but no such relation was observed with the oxygen partial pressure. These results suggest that the brain may be more resistant to oxygen free radicals as compared with the lungs, and that oxygen toxicity following HBO may be affected by factors other than oxygen free radicals.

활성 산소종과 반응한 솔잎이 Free radical에 미치는 효과

김수민,조영석 경산대학교 생명자원개발연구소 1999 생명자원과 산업 Vol.3 No.-

본 연구는 식품뿐만이 아니라 생체에 악영향을 미치는 산화반응 또는 radical 반응 전반에 걸쳐 반응성을 억제할 수 있는 항산화성 물질을 찾고자, 발암 및 노화를 방지하는 기능이 있다고 알려진 솔잎추출물로 각종 산소종과의 반응성을 검토한 결과 iron이 존재하지 않는 상태의 산화작용에 대해서는 솔잎추출물의 뚜렷한 작용이 나타나지 않았으며, free radical 반응에 중요한 매개체가 되는 ?? ion이 존재하는 산화반응에 솔잎추출물은 강력한 binding 효과를 나타내었다. 그러나, ??가 존재하는 산화반응에는 ?? ion이 존재하는 산화반응에서 보다 그 반응성이 매우 낮았으며, 이에 솔잎 추출물의 작용도 효과가 나타나지 않았다. ?? ion에 대한 binding 효과가 인정되는 솔잎추출물의 항산화능력을 비교하기 위하여 iron chelating에 우수한 합성항산 화제인 EDTA, DTPA보다 더욱 뛰어난 ?? ion binding 효과를 나타내었다. This study was carried out to investigate the effect of Pine needle extract on lipid oxidation and free radical reaction in iron sources reacted with active oxygen species. The results were summarized as follow; The pine needle extracts didn't show a distinct effect on reduction of lipid oxidation if the iron ion didn't exist in oil emulsion. The pine needle extracts played role as a strong chelating agents to bind iron ion if Ferrous(??) exist in oil emulsion. Ferric iron(??) was lower effect than Ferrous iron(??) on free radical reaction in oil emulsion. And also, the ?? reacted with pine needle extract did not show distinct effect on free radical reaction, compared to ?? reacted with pine needle extract. Pine needle extracts were the most powerful ?? binding agents, compared to other strong synthetic antioxidants such as EDTA and DTPA.