Effect of irradiation on cytokine secretion and nitric oxide production by inflammatory macrophages

차희재,최영현,임상욱,고은지,강윤정,백경완,옥미선,송경섭,강혜주,금영삼,현진원,권택규,남선영 한국유전학회 2016 Genes & Genomics Vol.38 No.8

This study explored the effects of low-dose and high-dose irradiation on inflammatory macrophage cells, specifically inflammatory cytokine secretion and nitric oxide (NO) production after irradiation. To elucidate the effect of irradiation on active and inactive macrophages, we exposed LPS-treated or untreated murine monocyte/macrophage RAW 264.7 cell lines to low-dose to high-dose radiation (0.01–10 Gy).Weanalyzed the effects of irradiation onRAW 264.7 cell proliferation byMTT assays and analyzed cytokine secretion and NO production related to inflammation by ELISA assays. Low-to-high doses of radiation did not significantly affect the proliferation of LPS-treated or untreated RAW 264.7 cells. Pro-inflammatory cytokine IL-1ß was generally increased in RAW 264.7 cells at 3 days after radiation. Especially, IL-1ß was significantly increased in only high dose-irradiation (2 and 10 Gy irradiation) groups in LPSuntreated RAW264.7 cells but increased in both lowand high dose-irradiation groups (0.01–10 Gy) in LPS-treated RAW 264.7 cells at 3 days after irradiation.Whereas, the expression of IL-1ß was prolonged in high-dose irradiation group at 5 days after irradiation. The production of anti-inflammatory cytokine IL-10 did not change significantly at 3 days after radiation but was significantly reduced at 5 days after 10 Gy radiation. The effect of irradiation on the secretion of IL-1ß and IL-10 was not significantly different between RAW264.7 cells treated or not treated with LPS. The effect of irradiation on NO secretion by RAW 264.7 cells showed a specific pattern. NO was produced after low-dose irradiation but reduced in a high-dose irradiation group at 3 days after irradiation. However, NO production was not changed after low-dose irradiation and reduced at 5 days after high-dose irradiation. These results showed that irradiation affected the inflammatory systemand regulatedNOproduction in both activated and inactivated macrophages through different regulation mechanisms, depending on irradiation dose.

Effect of gamma irradiation on Korean traditional multicolored paintwork

Yoon, Minchul,Kim, Dae-Woon,Choi, Jong-il,Chung, Yong-Jae,Kang, Dai-Ill,Hoon Kim, Gwang,Son, Kwang-Tae,Park, Hae-Jun,Lee, Ju-Woon Elsevier 2015 Radiation physics and chemistry Vol.115 No.-

<P><B>Abstract</B></P> <P>Gamma irradiation can destroy fungi and insects involved in the bio-deterioration of organic cultural heritages. However, this irradiation procedure can alter optical and structural properties of historical pigments used in wooden cultural heritage paintings. The crystal structure and color centers of these paintings must be maintained after application of the irradiation procedure. In this study, we investigated the effects of gamma irradiation on Korean traditional multicolored paintwork (Dancheong) for the preservation of wooden cultural heritages. The main pigments in Korean traditional wooden cultural heritages, <I>Sukganju</I> (Hematite; Fe<SUB>2</SUB>O<SUB>3</SUB>), <I>Jangdan</I> (Minium; Pb<SUB>3</SUB>O<SUB>4</SUB>), <I>Whangyun</I> (Crocoite; PbCrO<SUB>4</SUB>), and <I>Jidang</I> (Rutile; TiO<SUB>2</SUB>), were irradiated by gamma radiation at doses of 1, 5, and 20kGy. After irradiation, changes in Commision Internationale d’Eclairage (CIE) color values (<I>L</I>*, <I>a</I>*, <I>b</I>*) were measured using the color difference meter, and their structural changes were analyzed using X-ray diffraction (XRD) analysis. The slightly change in less than 1 <I>dE</I>* unit by gamma irradiation was observed, and structural changes in the Dancheong were stable after exposure to 20kGy gamma irradiation. In addition, gamma irradiation could be applied to painted wooden cultural properties from the Korean Temple. Based on the color values, gamma irradiation of 20kGy did not affect the Dancheong and stability was maintained for five months. In addition, the fungicidal and insecticidal effect by less than 5kGy gamma irradiation was conformed. Therefore, the optical and structural properties of Dancheong were maintained after gamma irradiation, which suggested that gamma irradiation can be used for the preservation of wooden cultural heritages painted with Dancheong.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Effects of gamma irradiation on the Dancheong were evaluated. </LI> <LI> We confirmed that optical and structural properties of Dancheong were maintained. </LI> <LI> Irradiation can contribute the decontamination for wooden cultural heritages. </LI> <LI> It also can be used for preservation of painted-wooden cultural heritages. </LI> </UL> </P>

Neutron irradiation performance of Zircaloy-4 under research reactor operating conditions

Jin, Hyun Ju,Kim, Tae Kyu Elsevier 2015 Annals of nuclear energy Vol.75 No.-

<P><B>Abstract</B></P> <P>Since more than 60% of operating research reactors in the world are over 40years old, aging management for their core structural components is one of the key issues faced to ensure life extension, reliability, and safe operation of these research reactors. To manage the aging process, a prediction for the irradiation behavior of the structural materials of the major components is required to avoid unplanned outages, and to enhance the safe and reliable operation. For this, the establishment of improved material properties for irradiated core structural materials in research reactors is essential. Zircaloy-4 is widely used as nuclear core structural materials such as fuel cladding and guide tubes due to its very low absorption cross-section of thermal neutrons, good mechanical property and corrosion resistance. In research reactors, Zircaloy-4 is subjected to relatively high neutron fluence and low temperature conditions compared with commercial reactors. Under such operating conditions, irradiation behaviors such as irradiation-induced dislocation, microstructural changes and irradiation growth in Zircaloy-4 were reviewed. In addition, irradiation-induced mechanical property changes obtained by a post irradiation examination were examined. This review will assist in the design as well as understanding of the irradiation behavior of Zircaloy-4 based core components in research reactors.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Neutron irradiation performance of Zircaloy-4 in research reactor environment. </LI> <LI> Microstructure evolution during irradiation. </LI> <LI> Characteristics of irradiation growth. </LI> <LI> Irradiation induced mechanical property change is examined. </LI> </UL> </P>

고용량 방사선 조사 후 골육종 세포주(Saos-2)의 아포프토시스 발생

김재도,정소학,홍영기,최장석,Kim, Jae-Do,Chung, So-Hak,Hong, Young-Gi,Choi, Jang-Seok 대한근골격종양학회 1999 대한골관절종양학회지 Vol.5 No.1

A single fraction of 50 Gy extracorporeal irradiation, as a modality of limb-sparing operation, has been used to achieve tumor necrosis in osteosarcoma. Although this modality of radiation therapy preserving the mobility of a joint is commonly practiced, the precise knowledge on the radiobiological response of osteosarcoma cell has remained to be elucidated. We therefore observed whether a single high dose irradiation caused apoptosis in osteosarcoma cells and whether the commitment to apoptosis was associated with cell kinetics. We also investigated radiation dose response along the time course for development of apoptosis following single high dose irradiation. The morphologic change in apoptosis was observed by fluorescence with Hoechst 33258 and the degree and the fraction of cells by flow cytometry. Irradiation of osteosarcoma cells with 10, 30 and 50 Gy resulted in chromatin condensation and apoptotic body formation. The degree of apoptosis in osteosarcoma cells was $29.5{\pm}3.56%$, $39.9{\pm}4.83%$ at 24 and 48 hours after 10 Gy irradiation ; $41.1{\pm}3.93%$, $66.9{\pm}5.21%$ at 24 and 48 hours after 30 Gy irradiation ; and $48.0{\pm}3.69%$, $75.6{\pm}4.65%$ at 24 and 48 hours after 50 Gy irradiation. The fraction of cells in cell-cycle kinetic was $39.2{\pm}4.3%$ in G2/M, $22.1{\pm}4.65%$ in G1 at 24 hours after 10 Gy irradiation ; $51.0{\pm}4.3%$ in G2/M, $20.4{\pm}4.7%$ in G1 at 48 hours after 10 Gy irradiation ; $40.3{\pm}3.9%$ in G2/M, $26.1{\pm}4.7%$ in G1 at 24 hours after 30 Gy irradiation ; $59.2{\pm}3.9%$ in G2/M, $5.9{\pm}5.1%$ in G1 at 48 hours after 30 Gy irradiation ; and $44.3{\pm}4.2%$ in G2/M, $21.1{\pm}3.5%$ in G1 at 24 hours after 50 Gy irradiation. The fraction of cells at 48 hours after 50 Gy irradiation could not be observed because of irradiation induced cell death of most of cells. All values for irradiated cells showed accumulation in G2/M phase and reduction in G1 phase, irrespective of irradiation dose. The results suggest that a single fraction of high dose irradiation with 50 Gy results in accumulation of cells at G2/M phase, leading to apoptosis.

PMA 처리한 인간 불멸화 상피세포에서 청색 및 적색 광 조사에 따른 활성산소 제거에 대한 비교 연구

한세우,고영종,정진안,김지은,김인애,임원봉,임회순,박지일,김미숙,김서연,김옥준,최홍란 대한구강악안면병리학회 2009 대한구강악안면병리학회지 Vol.33 No.1

Intracellular reactive oxygen species(ROS) produced in a various pathologic state was known to intermediate many cellular response such as inflammation. Recently, low level light irradiation by HeNe laser used in many clinical field could improve inflammatory state by scavenging intracellular ROS through photo-detachment/dissociation process. The purpose of this study is to investigate the differential effects of blue and red light irradiation on ROS scavenging effects. Immortalized human oral keratinocyte HaCat cells were used. Phorbol 12-myristate 13-acetate(PMA) was treated for inflammation. Red(635nm) and blue(470nm) light irradiation was carried out. To asses the intracellular ROS by light irradiation, confocal microscopic and flow cytometric assay with DCF fluorescence for total ROS and ESR spectrometry of DMPO-O2- for superoxide anion were caried out. And microarray was performed for mRNA expression level. Released intracellular total ROS in PMA treated HaCat cell lines was dissociated efficiently by red light irradiation, while blue light irradiation did not. Rather, blue light irradiation increased ROS formation. For superoxide anion generated the first synthetic form of ROS, red light irradiation reduced its amount but blue light irradiation did not. In the mRNA expression in line with cyclooxygenase(COX) pathway, prostagrandin endoperoxide synthase 1(PTGS 1), prostagrandin endoperoxide synthase 2(PTGS 2) and phospholipase A2(PLA2) were increased by both light irradiation and they were decreased as time flows. And genes associated with ROS releasing, mRNA expressions of tumor necrosis factor receptor (TNFR) and interleukin 1beta(IL1B) were increased by 1 hour red light irradiation but did not by blue light irradiation. As a result, red and blue light irradiation showed different response in affecting the level of ROS. These findings indicate that red light rather than blue light is more useful for anti-inflammation in clinical field.

PMA 처리한 인간 불멸화 상피세포에서 청색 및 적색 광 조사에 따른 활성산소 제거에 대한 비교 연구

한세우,고영종,정진안,김지은,김인애,임원봉,임회순,박지일,김미숙,김서연,김옥준,최홍란 대한구강악안면병리학회 2009 대한구강악안면병리학회지 Vol.33 No.1

Intracellular reactive oxygen species(ROS) produced in a various pathologic state was known to intermediate many cellular response such as inflammation. Recently, low level light irradiation by HeNe laser used in many clinical field could improve inflammatory state by scavenging intracellular ROS through photo-detachment/dissociation process. The purpose of this study is to investigate the differential effects of blue and red light irradiation on ROS scavenging effects. Immortalized human oral keratinocyte HaCat cells were used. Phorbol 12-myristate 13-acetate(PMA) was treated for inflammation. Red(635nm) and blue(470nm) light irradiation was carried out. To asses the intracellular ROS by light irradiation, confocal microscopic and flow cytometric assay with DCF fluorescence for total ROS and ESR spectrometry of DMPO-O2 - for superoxide anion were caried out. And microarray was performed for mRNA expression level. Released intracellular total ROS in PMA treated HaCat cell lines was dissociated efficiently by red light irradiation, while blue light irradiation did not. Rather, blue light irradiation increased ROS formation. For superoxide anion generated the first synthetic form of ROS, red light irradiation reduced its amount but blue light irradiation did not. In the mRNA expression in line with cyclooxygenase(COX) pathway, prostagrandin endoperoxide synthase 1(PTGS 1), prostagrandin endoperoxide synthase 2(PTGS 2) and phospholipase A2(PLA2) were increased by both light irradiation and they were decreased as time flows. And genes associated with ROS releasing, mRNA expressions of tumor necrosis factor receptor (TNFR) and interleukin 1beta(IL1B) were increased by 1 hour red light irradiation but did not by blue light irradiation. As a result, red and blue light irradiation showed different response in affecting the level of ROS. These findings indicate that red light rather than blue light is more useful for anti-inflammation in clinical field

수 종의 상피기원 종양 세포주에서 방사선 조사와 표피성장인자 투여에 따른 세포 주기의 변화와 apoptosis 유발에 관한 연구

한원정,허민석,이삼선,최순철,박태원 대한구강악안면방사선학회 2000 Imaging Science in Dentistry Vol.30 No.1

Purpose : This study was aimed to evaluate the cell cycle arrest and apoptosis induction after irradiation and epidermal growth factor(EGF) treatment in three human epithelial tumor cell lines (A43l, Siha, KB). Materials and Methods : Single irradiation of 2, 5 and 10 Gy was done on three cell lines with 5.38 Gy/min dose rate using Cs-137 irradiator at room temperature. Also, EGF of 10 ng/ml was added immediately after 10 Gy irradiation. Cell growth was evaluated by counting the living cell number using a hemocytometer at 1 day, 2 days, 3 days, 4 days and 5 days after irradiation. Cell cycle arrest and apoptosis induction were assayed with the flow cytometry at 8 hours, 12 hours, 1 day, 2 days, 3 days, 4 days and 5 days after irradiation. Results : Growth of irradiated three cell lines were inhibited in proportion to radiation dose, EGF treatment after irradiation showed various results according to cell lines. On all cell lines, G2 arrest was detected after 8 hours and maximized after 12 hours or 1 day. Amount of G2 arrest was positively dose dependent. However, EGF showed no significant change on G2 arrest. G2 arrest was recovered with time at 2 Gy and 5 Gy irradiation. However, at 10 Gy irradiation, G2 arrest was continued. Apoptosis was detected at 10 Gy irradiation. On EGF treated group after irradiation, A431 and Siha cell lines showed slightly increased apoptosis but there was no statistically significant difference. KB cell line showed no marked change of apoptosis induction. Conclusion : Irradiation effects on cell cycle arrest and apoptosis induction in three human epithelial tumor cell lines, however epidermal growth factor doesn't effect on. (Korean J Oral Maxillofac Radiol 2000; 30: 71-79)

쥐 섬유육종에서 베타카로틴과 방사선조사 병용의 항종양 효과

권형철(Hyoung-Cheol Kwon),양문식(Moon-Sik Yang) 대한방사선종양학회 2000 Radiation Oncology Journal Vol.18 No.2

목 적 : 베타카로틴과 방사선조사의 병용효과에 관한 평가를 목적으로, 베타카로틴을 병용한 경우 방사선조사 단독의 경우 보다 세포독성의 차이는 어떠하며, 또한 쥐 섬유육종에서 두 군간의 종양성장의 지연 정도에 어떠한 차이가 있는가를 관찰하고자 본 연구를 시행하였다. 대상 및 방법 : 2% 베타카로틴 유제를 2 mg/ml 으로 만든 다음 단계적으로 희석하여 사용하였으며, 섬유육종세포와 태생 5∼6주의 C3H/N의 실험쥐를 이용하였다. 방사선조사는 6 MV 선형가속기를 이용하였고, 세포내 독성은 쥐 섬유육종세포의 생존을 감소시키는 능력으로 평가하였으며, 베타카로틴 2 mg/ml을 방사선조사 1시간 전 섬유육종세포주에 접촉시켰다. 종양성장 지연 실험을 위하여 베타카로틴과 방사선조사 병용군(n=6)과 방사선조사 단독군(n=5)으로 분류하였으며, 베타카로틴 20 mg/kg을 방사선조사 30분전 섬유육종이 접종된 쥐의 복강내 일회 주사하였고, 방사선조사량은 20 Gy를 주었다. 종양용적은 장경×장경×장경/2 (mm3) 공식을 사용하였으며, 2∼3일 마다 측정하였다. 결 과 : 섬유육종세포에 베타카로틴 0.002, 0.02, 0.2, 2 mg/ml 농도액을 1시간 동안 접촉 후 얻은 각각 생존분율은 0.69±0.07, 0.59±0.08, 0.08±0.008 및 0.02±0.006이었다. 그리고 방사선조사 1시간 전 섬유육종세포에 베타카로틴 2 mg/ml을 접촉한 후 조사량 2, 4, 6 및 8 Gy에서 얻은 각각의 생존분율은 0.13±0.05, 0.03±0.005, 0.01±0.002 및 0.009±0.0008이었으며 방사선조사 단독군의 경우 동일 조사량에서 얻은 생존분율은 각각 0.66±0.05, 0.40±0.04, 0.11±0.01 및 0.03±0.006으로 나타났다(p<0.05). 종양성장의 지연정도를 나타내는 실험에서 섬유육종을 쥐에 접종한 후 종양의 용적이 1,000 mm3 에 달하는 기간은 베타카로틴 병용군과 방사선조사 단독군에서 각각 18일과 19일로 나타났다(p>0.05). 결 론 : 쥐 섬유육종세포에 베타카로틴을 접촉한 경우 세포독성이 나타났으며, 베타카로틴 농도 증가에 따라 세포독성도 증가하였다. 그리고 쥐 섬유육종세포의 세포독성은 베타카로틴 병용군에서 방사선조사 단독군의 경우 보다 부가적으로 증가하였으며, 두 군간에 통계학적으로 현저한 차이를 보였다. 그러나 쥐 섬유육종 성장 지연정도에 있어서 베타카로틴 병용군과 방사선조사 단독군간의 통계학적으로 뚜렷한 차이는 없었다. Purpose :To investigate whether combined beta- carotene with X- irradiation has more enhanced radition response than X- irradiation or not, we performed a experiment about in vitro cytotoxicity of beta- carotene and/or X- irradiation in the fibrosarcoma cells, tumor growth delay of combined beta- caroten with/or X- irradiation in the mouse fibrosarcoma. Materials and Methods :2% emulsion of beta- carotene was serially diluted and used. X- irradiation was given by 6 MeV linear accelerator. The cytotoxicity of beta- carotene in vitro was evaluated from clonogenic assay. To compare the cytotoxicity between combined beta- carotene with X- irradiation and X- irradiation group, 2 mg/ml of beta- carotene was contacted to fibrosarcoma (FSaII) cells for 1 hour before X- irradiation. For the tumor growth delay, single 20 Gy was given to FSaII tumor bearing C3H/N mice whic was classified as beta- crotene with X- irradiation group (n=6) and X- irradiation alone group (n=5). 0.2 ml of 20 mg/kg of beta- carotene were i.p. injected to mice 30 minute before X- irradiation in the beta- crotene with X- irradiation group. The tumor growth delay defined as the time which reach to 1,000 mm3 of tumor volume. Result : (1) Cytotoxicity in vitro; 1) survival fraction at beta- carotene concentration of 0.002, 0.02, 0.2 and 2mg/ml were 0.69±0.07, 0.59±0.08, 0.08±0.008 and 0.02±0.006, respectively. 2) each survival fraction at 2, 4, 6 and 8 Gy in the 2 mg/ml of beta- carotene +X- irradiation group were 0.13±0.05, 0.03±0.005, 0.01±0.002 and 0.009±0.0008, respectively. But each survival fraction at same irradiation dose in the Xirradiation group were 0.66±0.05, 0.40±0.04, 0.11±0.01 and 0.03±0.006, respectively(p<0.05). (2) The time which reach to 1,000 mm3 of tumor volume of beta- carotene + X- irradiation group and X- irradiation alone group were 18, 19 days, respectively(p>0.05). Conclusion :The contact of beta- caroten to FSaII cells showed mild cytotoxicity which was increased according to concentration. The cytotoxicity of combined beta- carotene with X- irradiation more increased than that of X- irradiation, additionaly. And there was significant difference of cytotoxicity between two groups. But there were no significant difference of the growth delay of fibrosarcoma between two groups.

SOD 전처치가 방사선조사후 흰쥐 넙다리곧은근의 미세구조 변화에 미치는 영향

백두진,한규희,정호삼 대한해부학회 1998 Anatomy & Cell Biology Vol.31 No.4

Effects of 2-deoxy-D-glucose and quercetin on the expression of osteonectin and osteopontin during the differentiation of irradiated MC3T3-E1 osteoblastic cells

류수경,고광준,김경아 대한영상치의학회 2008 Imaging Science in Dentistry Vol.38 No.4

Purpose : To characterize the effects of 2-deoxy-D-glucose (2-DG) and quercetin (QCT) on gene expression of osteonectin (ON) and osteopontin (OP) in irradiated MC3T3-E1 cells. Materials and Methods : When MC3T3-E1 osteoblastic cells had reached 70-80% confluence, cultures were transferred to a differentiating medium supplemented with 5 mM 2-DG or 10 μM QCT and then irradiated with 2, 4, 6, and 8 Gy. At various times after irradiation, the cells were analyzed for the expression of bone mineralization genes such as ON and OP. Results : The mRNA expression of both ON and OP was increased according to the culture time in the differentiation medium, and the increase of the genes peaked at 14 days after the differentiation induction. In the case of OP, the increase of mRNA expression was maintained to 28 days after the differentiation, while the mRNA level of ON was reduced to the basal level at the same time. Irradiation adding 2-DG showed a significant peak value in the expression pattern of ON at 4 Gy 7 days after irradiation. Irradiation adding QCT increased the mRNA expression of ON and OP in a dose-dependant manner, but irradiation adding 2-DG did not show any differences between the control and experiments 14 days after irradiation. Irradiation adding QCT increased significantly the expression patterns of ON 21 days after irradiation. Conclusion : The results showed that QCT acted as a radiosensitizer in the gene expression of ON and OP during differentiation of the late stage of irradiated MC3T3-E1 osteoblastic cells in vitro. (Korean J Oral Maxillofac Radiol 2008; 38 : 195-202) Purpose : To characterize the effects of 2-deoxy-D-glucose (2-DG) and quercetin (QCT) on gene expression of osteonectin (ON) and osteopontin (OP) in irradiated MC3T3-E1 cells. Materials and Methods : When MC3T3-E1 osteoblastic cells had reached 70-80% confluence, cultures were transferred to a differentiating medium supplemented with 5 mM 2-DG or 10 μM QCT and then irradiated with 2, 4, 6, and 8 Gy. At various times after irradiation, the cells were analyzed for the expression of bone mineralization genes such as ON and OP. Results : The mRNA expression of both ON and OP was increased according to the culture time in the differentiation medium, and the increase of the genes peaked at 14 days after the differentiation induction. In the case of OP, the increase of mRNA expression was maintained to 28 days after the differentiation, while the mRNA level of ON was reduced to the basal level at the same time. Irradiation adding 2-DG showed a significant peak value in the expression pattern of ON at 4 Gy 7 days after irradiation. Irradiation adding QCT increased the mRNA expression of ON and OP in a dose-dependant manner, but irradiation adding 2-DG did not show any differences between the control and experiments 14 days after irradiation. Irradiation adding QCT increased significantly the expression patterns of ON 21 days after irradiation. Conclusion : The results showed that QCT acted as a radiosensitizer in the gene expression of ON and OP during differentiation of the late stage of irradiated MC3T3-E1 osteoblastic cells in vitro. (Korean J Oral Maxillofac Radiol 2008; 38 : 195-202)