방사선조사 흰쥐의 간 및 비장조직중의 Superoxide Dismutase 활성도에 대한 연구

이병래,정두영,양종대,차종희 조선의대 1985 The Medical Journal of Chosun University Vol.10 No.1

고농도 포도당이 흰쥐 인슐린 종 세포의 superoxide dismutase 발현에 미치는 영향

이재우,이병래 조선대학교 2000 The Medical Journal of Chosun University Vol.25 No.2

Background: Reactive oxygen species are produced under diabetic condition and possibly cause various forms of tissue damage in patients with diabetes mellitus. The aim of this study is to examine the effects of high glucose on the gene expression of superoxide dismutase(SOD) isozymes in rat insulinoma cells line. Methods: Rat insulinoma(RINm5F) cells were cultured with RPMI 1640 media containing high glucose(22.2mM), or normo-glucose(5.6mM) for 3 days and for 21 days. The SOD activities were determined by spectrophotometric assay and nitroblue tetrazolium (NBT) stain. The levels of SOD mRNA were quantified by reverse transcriptase-polymerase chain reaction (RT-PCR). Results: The CuZn-SOD activity was decreased but Mn- SOD activity was increased significantly in RINm5F cells which cultured with high glucose(22.2mM) media for 3 days. In RINm5F cells, the Mn-SOD mRNA content was also increased by high glucose, but the level of CuZn-SOD mRNA was not changed by gluose concentration for 3 days culture. Both CuZn-SOD and Mn- SOD activity were increased significantly when cultured with high glucose media (22.2mM) for 21 days. The levels of Mn-SOD and CuZn-SOD mRNA were increased in RINm5F cells cultured with in high glucose media for 21 days. Conclusion: These results suggest that in RINm5F cells, both CuZn-SOD and Mn-SOD gene expressions maybe increase by high glucose and decrement of CuZn-SOD activity, in cells of short term culture with high glucose contained media may due to the protein modification.

가토 적혈구에서 철이온에 의해 유도되는 지질과산화에 미치는 paraquat의 영향

이병래,김명철,차종희 朝鮮大學校 附設 醫學硏究所 1991 The Medical Journal of Chosun University Vol.16 No.2

The participation of superoxide in initiating tissue damage from admini stration of a xenobiotics is best illustrated by paraquat intoxication, in which it is known that one electron reduction of para quat leads to the formation of radical species which react with molecular oxygen to give superoxide. Although it has also been argued that superoxide itself is not toxic, the shunting of electrons into the futile reduction of oxygen to hydrogen peroxide can be considered as a toxic process. In the present study, the effect of paraquat on lipid peroxidation in rabbit erythrocyte was investigated, Incubation of a buffered aerobic mixture of erythrocyte in the presence of Fe^(2+) and glucose or NADH resulted in the formation of lipid peroxide as determined as malonydialdehyde. Paraquat inhibited Fe^(2+) induced lipid peroxidation in rabbit erythrocyte. In contrast, NADH and glucose stimulate the Fe^(2+) induced lipid peroxidation of erythrocyte in the presence of paraquat and the levels of erythrocyte NADH and NADPH were decreased by paraquat. These results suggest that the inhibitory effect of paraquat on FeU induced lipid peroxidation in rabbit erythrocyte is mainly due to a decrease in the level of NADH and NADPH.

만성폐쇄성 폐질환 환자에서 적혈구 항산화효소의 변화

국기용,이만재,조희충,임영국,박일,김원식,이승일,이병래 朝鮮大學校 附設 醫學硏究所 1993 The Medical Journal of Chosun University Vol.18 No.1

Toxic oxygen free radicals have been implicated as important pathologic mediators in many clinical disordesrs. Enhandng the intracellular content of antioxidant enzymes can provide means of limiting biological damage caused by oxygen free radicals. Many investigations have examined the potential of various oxyradical scavengers to modulate oxyradical injury. The oxygen free radicals play a role in the pathogenesis of chronic obstructive pulmonary disease. To investigate the pulmonary oxyradical injury and the protective role of antioxidant enzymes, author measured the amount of thiobarbituric acid reactants, the activiteies of antioxidant enzymes (superoxide dismutase, glutathione peroxidase, and catalase) and the sulfhydryl groups in serum and red blood cells from the chronic obstructive pulmonary disease groups and the normal controls. The thiobarbituric acid reactant in chronic obstructive pulmonary disease groups was increased than the controls in both serum and red blood cells, and the superoxide dismutase activity in red blood cells was no statistically difference in both groups. But, the glutathione peroxidase and catalase activities were lowered in chronic obstructive pulmonary disease groups than the controls. The sulfhydryl groups in serum and red blood cells were no statistically difference in both groups. These results suggest that the increased thiobarbituric acid reactant in chronic obstructive pulmonary disease groups mean oxygen radical toxicity, and the decreased glutathione peroxidase and catalase activities may play a role in cellular injury.

칼로리량 급여 제한이 OLETF 흰쥐의 콜레스테롤량에 미치는 영향

박재윤,김태형,이병래,차종희 조선대학교 부설 의학연구소 2002 The Medical Journal of Chosun University Vol.27 No.1

본 실험은 식이 제한 급여에 따른 비만형 당뇨병 환자의 체중 및 혈중 지질량의 변화에 대한 연구의 일환으로 사람의 비만형 당뇨병과 유사한 병리소견을 나타내는 OLETF 흰쥐를 이용하여 칼로리 급여량 비 제한군, 칼로리 급여량 20% 제한군과 40% 제한군으로 나누어 12주간 사육한 후 체중과 혈당, 혈액 cholesterol 및 중성지질량을 측정하여 칼로리 급여량제한에 따른 혈액 지질량의 변화를 관찰하였다. 체중은 칼로리 급여량 20% 제한군에서는 체중증가 억제 비율이 LETO와 OLETF흰쥐가 비슷하였고, 칼로리 급여량 40% 제한군에서는 OLETF 흰쥐에서 체중 증가 억제가 더 크게 나타났다. 칼로리 급여량 제한에 의한 혈당량의 저하는 비만형 당뇨쥐인 OLETF 증 흰쥐가 LETO 흰쥐에 비하여 더 크게 나타났다. 칼로리 급여량 제한으로 OLETF 종 흰쥐에서 혈액 중성지질량, 총 콜레스테롤량 및 LDL- 콜레스테롤량이 감소되었고, HDL-콜레스테롤량은 증가되므로, 비만형 당뇨병환자에서 칼로리 제한 식사는 OLETF 종 흰쥐에서와 유사한 혈액 콜레스테롤량의 변화를 유발시켜서 혈관 합병증을 억제할 수 있을 것으로 예상된다. 이상의 실험결과로서 칼로리 급여량 제한에 의한 체중, 혈당, 혈액 총 콜레스테롤량, LDL-콜레스테롤 및 HDL-콜레스테롤량의 변화가 비당뇨 흰쥐보다 당뇨흰쥐에서 더 큰 변화가 있음이 확인되어 칼로리 제한 급여는 당뇨병에서 체중, 혈당, 콜레스테롤량의 조절에 유의한 것으로 생각되고, 칼로리 급여 20% 제한군과 40% 제한군 사이에서 체중, 혈당, 콜레스테롤량의 변화는 칼로리 제한량의 변화에 비하여 적기 때문에 적절한 체중, 혈당, 혈중 지질량의 유지를 위해서는 칼로리 급여량을 적절히 유지하는 것이 중요할 것으로 생각된다. 또한 칼로리 급여 제한에 의한 체중, 혈당, 혈중 중성지질과 콜레스테롤량의 감소는 혈관 합병증 발생이 예방 할 수 있을 것으로 추측되는데, 이에 대해서는 계속적인 연구가 있어야 할 것으로 생각된다. Objective: To determine the effects of caloric restriction on blood cholesterol levels of obese type 2 diabetes, we measured blood glucose and cholesterol level in Otsuka Long Evans Tokushima Fatty(OLETF) rats after caloric restriction. Methods: OLETF rats(obese diabetic rats) and Long Evans Tokushima Otsuka(LETO) rats(nondiabetic control rats) were grouped into 3 groups, control group, 20% caloric restriction(20% CR) group and 40% caloric restriction(40% CR) group, respectively. The blood glucose and cholesterol level in OLETF and LETO rats were determined after 12 weeks caloric restriction. The levels of triglyceride(TG) and cholesterol(total cholesterol, LDL-cholesterol, HDL-cholesterol) in blood of rats were determined by automatic biochemical analyser(Hitachi 747 Japan). Results: Blood glucose level of LETO rats were decreased by 12% on 40% CR compared to control group but the differences of blood glucose levels between control group and 20% CR group was not observed. OLETF rats blood glucose levels were decreased by 20% in 40% CR group and by 15% in 20% CR group. The levels of blood triglyceride in 20% CR and 40% CR groups were decreased by 20%, 15% in LETO rats and by 37%, 32% in OLETF rats respectively. Total cholesterol level was not changed by dietary restriction in LETO rats, but in OLETF rats, total cholesterol levels was decreased significantly by caloric restriction. The HDL-cholesterol levels were increased while LDL-cholesterol levels were decreased by dietary restriction in both LETO and OLETF rats. Conclusion: These results suggested that OLETF rats may be more susceptive to changes of body weight and blood glucose, triglyceride and cholesterol levels than LETO rats by caloric restriction. Reduced LDL-cholesterol and triglyceride levels in diabetes by caloric restriction may have some roles on prevention of diabetic vascular complication.

흰쥐에서 paraquat독성에 미치는 tumor necrosis factor-α의 영향

차종희,박정수,양성훈,박재윤,이병래 조선대학교 2000 The Medical Journal of Chosun University Vol.25 No.2

Background : The toxicity of paraquat is due to the oxygen-derived radicals formed by the reaction of oxygen with bipyridylium radical cations. Tumor necrosis factor (TNF)-α induces the synthesis of mitochondrial manganese superoxide dismutase (Mn-SOD) and other protective proteins in a variety of tumor cells and these protein contribute to a protect tumor cells from free radical injury. In this study, the effects of TNF-α on paraquat-induced toxicity were investigated in rats. Methods : Experimental animal used was Sprague-Dawley male rat, body weight 150-200g. The paraquat toxicity was determined by survival rate. The SOD, catalse, and glutathione peroxidase(GSHPx) activity were spectrophtomerically determined by Crapo method, Aebi method and Flohe method, respectively. Results : The survival rate of paraquat-intoxicated rats was decreased by subctaneous injection of TNF-α. However, the survival rate of paraquat injected rats were increased by pretreatment of TNF-α, comparing to saline pretreatment. The liver SOD activity was increased significantly by TNF-α treatment, but kidney and lung enzyme was unaltered in TNF-α treated-rats, comparing to normal control rats. The increment of liver SOD activity by TNF-α treatment was dose- and time-dependant SOD activity of kidney and lung of rats were not changed by TNF-α injection. The activities of catalase and glutathione peroxidase and contents of thiobarbituric acid reactive substance(TBARS) in liver were not changed by TNF-α treatment. Conclusion : From these result, TNF-α potentiate paraquat toxicity in rats but TNF-α pretreatment reduce paraquat induced toxicity. Protective effect may be resulted from the increased liver SOD activities by TNF-α.

Paraquat 투여 백서에서 Cyclophosphamide와 Dexamethasone의 치료 효과

신정철,신양수,유병전,임현성,정태흥,박찬국,오금탁,이병래 朝鮮大學校 附設 醫學硏究所 1991 The Medical Journal of Chosun University Vol.16 No.1

Paraquat (N,N'-dimethy 1-4,4'bipyrydinium : Gramoxone^(R), a widely used herbicide, is extremely toxic to all green plants and many eukaryotic organisms. In human, paraquat intoxication leads to multiple organ failure, it preferentially damages the lung, kidney and liver and may result in death. The participation of superoxide in initiating tissue damage by administration of paraquat is well known in which one electron reduction of paraquat leads to the formation of radical species which react with molecular oxygen to give superoxide. This process initiates immunological changes with the activation of neutrophilic leukocytes, which is related to further production of superoxide. In this experiment, therapeutic effects of cyclophosphamide and dexamethasone on blood cell count, malondialdehyde as a indicator of lipid peroxidation, and antioxidant enzymes levels in the lung, liver and kidney of paraquat intoxicated rats were inbestigated. The WBC count were significantly decreased by combined treatment of cyclophosphamide and dexamethasone in paraquat intoxicated rats at 3 and 5 days but RBC count were not changed. The contents of malondialdehyde were significantly decreased in the lung and Kidney by combined treatment at 10 days after paraquat intoxication. The catalase acitvities were markedly increased in the lung and liver of rats at 10 days after paraquat intoxication, and superoxide dismutase activities showed minimal changes in the lung, liver and kidney of rats by combined treatment. These results suggest that paraquat induced tissue damage in rats can be redeced by combined administration of cyclophosphamide and dexamethasone.

녹차 및 결명자 추출물의 교정용 브라켓과 치면 사이의 경계부에서 분리된 mutans streptococci에 대한 항균작용

임성훈,서정순,윤영주,김광원,유소영,김화숙,국중기,이병래,차종희,박재윤 대한치과교정학회 2003 대한치과교정학회지 Vol.33 No.5

본 연구는 치아우식증에 관련된 Mutans streptococci 표준균주 및 임상분리 균주의 성장억제를 유도할 수 있는 천연 생약추출물을 검색하기 위하여 실시되었다. 녹차추출물은 CHMC-2032를 사용하였으며, 결명자 추출물은 50% 에탄올을 이용하여 얻었다. 이들 추출물의 Mutans streptococci 표준균주 및 교정환자들에서 브라켓과 치면 사이의 경계부에서 분리된 각각 10 균주씩의 S. mutans 및 S. sobrinus에 대한 최소성장억제농도를 액체배지 희석법으로 구하였다. 그 결과 CHMC-2032의 S. mutans 및 S. sobrinus의 표준 균주, S. sobrinus의 대부분 임상분리 균주에 대한 최소성장억제농도는 5 mg/ml이었다. 그러나 결명자 추출물에 의한 S. mutans 및 S. sobrinus에 대한 세균 성장억제 효과는 미미하였다. 본 연구 결과 치아우식증의 예방적 측면에서 결명자차보다는 녹차를 마시는 것이 유리하며, CHMC-2032를 이용하여 구강양치용액을 제조하여 사용할 경우 교정환자를 포함하여 대부분의 사람에게서 치아우식증 예방 효과가 있으리고 추정된다. Mutans streptococci is the major causative factor in dental caries. Especially, orthodontic patients with fixed appliance are a risk group for dental caries. Because fixed appliances attached on teeth may change the environment of dental plaque, the enamel decalcification or dental caries around the bracket and band is a major side effect of orthodontic treatment. The aim of this study was to search plant extracts that have antimicrobial effect on mutans streptococci. Seed-extract of Casio tora were prepared with ethanol and CHMC-2032, the leaf-extracts from Camellia sinensis extract, was obtained extract, 2 type strains and 20 clinical isolates of mutans streptococci isolated from the interface between orthodontic brackets and tooth surfaces in the orthodontic patients were used in this study. The minimal inhibitory concentration of CHMC-2032 was 5 mg/ml on the S. mutans KCTC 3065, S. sobrinus KCTC 3088, and 8 clinical isolates of S. sobrinus. However, there was no antibacterial effect of seed-extract of C. tora on mutans streptococci. These data suggest that green tea may be more effective than the tea prepared from C. tora in the prevention of enamel decalcification or dental caries around brackets.

Hypoglycemic effect of hot-water and ethanol extract of Pleurotus nebrodensis

Byoung-Rai Lee,Hyun-Jae Shin,Wol-Suk Cha 한국생물공학회 2007 한국생물공학회 학술대회 Vol.2007 No.10

사람 폐암 세포주에서 시스플라틴이나 독소루비신의 세포독성에 미치는 녹차 추출물의 영향

이병래(Byoung-Rai Lee),박재윤(Jae-Yoon Park),박평심(Pyoung-Sim Park) 한국식품영양과학회 2011 한국식품영양과학회지 Vol.40 No.5

항암 화학요법제의 항암작용을 증가시키거나, 부작용을 감소시켜 항암 치료를 효과적으로 할 수 있는 항암치료 보조제(modulator)에 대한 개발의 일환으로 녹차 추출물의 이용 가능성을 추정하기 위하여 사람 폐암 세포주인 A549 세포를 배양하여 시스플라틴과 독소루비신의 항암성에 미치는 녹차 추출물과 EGCG의 영향을 비교 관찰하였다. A549 세포에 독성을 나타나는 농도는 녹차 추출물 400 ㎍/mL, EGCG 300 ㎍/mL, 시스플라틴 10 ㎍/mL 및 독소루비신 8 ㎍/mL로, 녹차 추출물이 세포독성을 나타내는 농도는 시스플라틴이나 독소루비신에 비하면 낮았다. A549 세포에서 시스플라틴 10 ㎍/mL 이상의 농도에서 세포활성이 감소되었고, EGCG나 녹차 추출물 100 ㎍/mL를 첨가하면 시스플라틴 6 ㎍/mL 이상의 농도에서 세포활성이 감소되어 EGCG나 녹차 추출물 첨가로 시스플라틴의 세포독성이 증가되었다. A549 세포에서 독소루비신 8 ㎍/mL 이상의 농도에서 세포활성이 감소되었고, EGCG나 녹차 추출물 100 ㎍/mL를 첨가하면 독소루비신 4 ㎍/mL 이상의 농도에서 세포활성이 감소되어 EGCG나 녹차 추출물 첨가로 독소루비신의 세포독성이 증가되었다. A549 세포에서 녹차추출물 투여 후 p53 및 caspase-3에 대한 Western blot을 시행한 결과 p53및 caspase-3의 유전자 발현이 증가되었다. 이상의 실험결과 녹차추출물은 광범위 항암제 시스플라틴이나 독소루비신의 세포독성을 증강시키는 효과가 있고, 녹차추출물에 의한 p53이나 caspase-3 등과 같은 세포자살유도 단백질의 발현 증가는 녹차추출물에 의한 세포독성 증강효과와 연관이 있을 것으로 추측된다. 녹차추출물의 시스플라틴이나 독소루비신 세포독성 증강효과는 항암화학요법제의 용량을 늘리지 않고 항암력을 증대시킬 수 있기 때문에 항암화학요법 보조제로서 이용될 수 있는 가능성이 높은 것으로 생각되며, 이러한 효과를 규명하기 위한 연구가 필요할 것으로 사료된다. Tea extract (TE) has been shown to have anti-tumor properties in a wide variety of experimental systems. We evaluated green tea extract (GTE) as a biochemical modulator for the antitumor activity of cisplatin and doxorubicin in the treatment of human lung cancer A549 cells. Cells were grown in RPMI-1640 medium supplemented with 10% (v/v) heat-inactivated fetal bovine serum and two antibiotics (100 units/mL penicillin and 100 ㎍/mL streptomycin). Two types of TE, epigallocatechin galate (EGCG) and GTE, were used in this experiment. The cells were seeded at 1×10⁴ cells/well in the RPMI-1640 media with or without TE (100 ㎍/mL) and then treated with different concentrations of doxorubicin (0~14 ㎍/mL) or cisplatin (0~35 ㎍/mL). After incubation in 5% CO₂ at 37℃ for 24 hr, cell viability was determined with a MTT assay. We used a Western blot to detect the influence of EGCG and GTE on the expression of p53 and caspase-3 genes in the A549 cells. A549 cell viability decreased to 15% with a 10 ㎍/mL concentration of cisplatin, and to 21% with a 8 ㎍/mL concentration of doxorubicin, as measured with the MTT assay. However, pre-treatment of the cells with EGCG (100 ㎍/mL) or GTE (100 ㎍/mL) resulted in decreased cell viability with 6 ㎍/mL of cisplatin and 4 ㎍/mL of doxorubicin. There was no apparent change in cell viability between EGCG or GTE administration in cisplatin- or doxorubicin-induced cytotoxicity in A549 cells. The levels of p53 and caspase-3 in the A549 cells increased with both EGCG and GTE treatment. We found that GTE could potentially affect cisplatin- or doxorubicin-induced cytotoxicity of A549 cells, which may be useful in the chemotreatment of cancer.