막지질 과산화와 간세포내 마이크로솜 및 리덕타제 기능과의 상관성에 관한 연구

박상열,조종후,Park, Sang-Youel,Cho, Jong-Hoo 대한수의학회 2004 大韓獸醫學會誌 Vol.44 No.2

The effects of membrane lipid peroxidation and retinyl palmitate on rat liver microsomal functions were investigated in vitro. Rat liver homogenates exposed to oxygen tension for 0, 3, 6, 9 or12 hours and lipid peroxidation levels were evaluated by the measurements of fluorescence intensity, malondialdehyde (MDA) and retinyl palmitate. The fluorescence intensity of homogenates and microsomes were elevated and retinyl palmitate concentrations were decreased. But the concentration of MDA was not affected to exposure time. Therefore, fluorescence intensity and retinyl palmitate concentration were used to analyze the correlation between lipid peroxidation and microsomal functions. To investigate the liver microsomal functions, the microsome was isolated from rat liver homogenates exposed to oxygen. The concentration of cytochrome P450 and the activity of NADPH-cytochrome P450 reductase in liver microsomes were gradually decreased with increasing the exposure time. The correlation between fluorescence intensity of microsomes showed a very high inverse correlation of -0.97 and -0.93, respectively. The decrease of cytochrome P450 concentration was due to the regeneration of cytochrome P450 to cytochrome P420. Also, the activities of cytochrome P450-dependent aminopyrine demethylase and benzpyrene hydroxylase of liver microsomes were gradually decreased with increasing the exposure time. The correlation with fluorescence intensity of microsome showed a high inverse correlation of -0.97 and -0.91, respectively. The retinyl palmitate concentrations of rat liver homogenates were decreased with increasing the exposure time. The decrease of retinyl palmitate concentration was followed by a low concentration of cytochrome P450 and activity of NADPH-cytochrome P450 reductase. The correlation indicated high direct correlation of 0.92 and 0.93, respectively. The decrease of retinyl palmitate concentration was also accompanied by the reduction of aminopyrine demethylase and benzpyrene hydroxylase activities. The correlation was analyzed a high direct correlation of 0.90 and 0.85, respectively. In conclusion, these studies have shown that the membrane lipid peroxidation of rat liver microsome proportionally decreased microsomal enzyme activities in vitro experiments.

TRAIL 유도 세포사멸에 있어서 IFN-γ의한 증가 기전 연구: IRF-1과의 관련성

박상열,설재원,이유진,강석진,김인식,강형섭,채준석,조종후,Park, Sang-Youel,Seol, Jae-Won,Lee, You-Jin,Kang, Seog-Jin,Kim, In-shik,Kang, Hyung-sub,Chae, Joon-seok,Cho, Jong-Hoo 대한수의학회 2004 大韓獸醫學會誌 Vol.44 No.2

Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) is a member of the TNF family and potent inducer of apoptosis. TRAIL has been shown to effectively limit tumor growth in vivo without detectable cytotoxic side effects. Interferon (IFN)-${\gamma}$ often modulates the anti-cancer activities of TNF family members including TRAIL. We previously reported that IFN-${\gamma}$ enhanced TRAIL-induced Apoptosis in HeLa cells without the unknown mechanism. In this study, we investigated whether IRF-1 involves in IFN-${\gamma}$-enhanced TRAIL-induced apoptosis. We exposed HeLa cells to IFN-${\gamma}$ for 12 hours and then treated with recombinant TRAIL protein. No apoptosis was induced in cells pretreated with IFN-${\gamma}$, and TRAIL only induced 30% apoptosis after 3 hours treatment. In HeLa cells pretreated with IFN-${\gamma}$, TRAIL induced cell death to more than 75% at 3 hours, showed that IFN-${\gamma}$-pretreatment enhanced HeLa cell death to TRAIL-induced apoptosis. To investigate the functional role of IRF-1 in IFN-${\gamma}$-enhanced TRAIL-induced apoptosis, IRF-1 was overexpressed by using an adenoviral vector AdIRF-1. IRF-1 overexpression increased apoptotic cell death and significantly enhanced apoptotic cell death induced by TRAIL when infected cells were treated with TRAIL. Our findings show that IFN-${\gamma}$ enhances TRAIL-induced apoptosis by IRF-1 in HeLa cells.

한우 무손상 적혈구의 superoxide 및 과산화수소 제거능력

조종후,박상열,Cho, Jong-hoo,Park, Sang-youel 대한수의학회 1998 大韓獸醫學會誌 Vol.38 No.2

The ability of bovine intact red blood cells to scavenge superoxide and hydrogen peroxide by superoxide dismutase, catalase and glutathione peroxidase was investigated. Intact red cells(up to 0.4%) suspensions did not inhibit ferricytochrome c reduction by superoxide in the superoxide generating system. On the other hand, intact red cell(0.4%) suspensions almost completely inhibit ferrocytochrome c oxidation by hydrogen peroxide. The ability of intact red cells to scavenge hydrogen peroxide was mainly attributed to either membrane bound catalase or glutathione peroxidase. The scavenge of hydrogen peroxide by 0.1~0.2% intact red cells showed a trend of dependence on mainly glutathione peroxidase. However, at blood cell concentration higher than 0.3%, the process depended upon peroxidase-independent scavengers like catalase. Enhancement of ferrocytochrome c oxidation by red cells treated with aminotriazole proved that the protection against hydrogen peroxide was due to catalase, while the protection in the presence of glutathione indicated scavenging effect of glutathione peroxidase against hydrogen peroxide.

Proteasome 억제에 의한 P53의 발현과 미토콘드리아 막 전압의 소실로 TRAIL에 저항하는 폐암세포의 사멸 강화

설재원,박상열,Seol, Jae-Won,Park, Sang-Youel 대한수의학회 2009 大韓獸醫學會誌 Vol.49 No.1

The ubiquitin-proteasome mediated protein degradation pathway plays an important role in regulating both cell proliferation and cell death. Proteasome inhibitors are well known to induce apoptosis in various human cancer cell lines. We investigated the effect of combined treatment with proteasome inhibitor and TRAIL, and a possible mechanism of the enhancing apoptosis by the both treatment, on TRAIL-resistant non-small cell lung cancer. A549 cells were exposed to the N-Acetyl-Leu-Leu-Norleu-al (ALLN) as a proteasome inhibitor and then treated with recombinant TRAIL protein. In A549 cells under proteasome inhibition conditions by pretreatment with ALLN, TRAIL treatment significantly decreased cell viability compared to that ALLN and TRAIL alone treatment. Also, the both treatment induced cell damage through DNA fragmentation and p53 expression. In addition, the combined treatment of both markedly increased caspase-8 activation, especially the exposure for 2 h, and Bax expression and induced the dissipation of mitochondrial transmembrane potential in A549 cells. Taken together, these findings showed that proteasome inhibition by ALLN enhanced TRAIL-induced apoptosis via DNA degradation by activated P53 and mitochondrial transmembrane potential loss by caspase-8 activation and bax expression. Therefore, our results suggest that proteasome inhibitor may be used a very effectively chemotherapeutic agent for the tumor treatment, especially TRAIL-resistant tumor cell.

연령증가에 따른 마이크로솜 막지질 과산화수준의 변화와 해독효소계의 관계

조종후,황대우,박상열,Cho, Jong-Hoo,Hwang, DaeWoo,Park, Sang-Youel 대한수의학회 2003 大韓獸醫學會誌 Vol.43 No.4

The studies were carried out on the correlation between microsomal lipid peroxidation level and drug metabolizing enzyme activities in rat liver microsomal suspensions on various ages (2-week-old, 2, 4, 8, and 12-month-old). The lipid peroxidation levels of liver homogenates tended to be elevated in a 4-month-old rat livers, but it was a little decreased in 8 and 12-month-old rat livers. The lipid peroxidation levels of microsomal suspension was not shown any significant differences by ages. Lipid peroxidation levels and microsomal cytochrome P450 and NADPH-cytochrome c reductase activity showed a direct correlation (r=0.72 and r=0.64), respectively. The activities of cytochrome P450-dependent aminopyrine-N-demethylase and benzpyrene hydroxylase in rat liver microsomes were increased by ages up to 8-month-old rats and maintained in 12-month-old rats. The correlation between lipid peroxidation levels and these cytochrome-dependent enzyme activities showed a high direct correlation (r=0.97 and r=0.81), respectively.

한우(韓牛) 송아지의 혈청단백분획(血淸蛋白分劃)에 관(關)한 연구(硏究)

김홍섭,이주묵,권오덕,박진호,박상열,이승옥,Kim, Heung-seop,Lee, Joo-mook,Kwon, Oh-deog,Park, Jin-ho,Park, Sang-youel,Lee, Sung-ok 대한수의학회 1997 大韓獸醫學會誌 Vol.37 No.2

120 heads of korean native calves were examined of serum protein by using cellulose acetate electrophoresis. During 20 days since the calves were purchased, serum protein, fibrinogen values and plasma : fibrinogen ratio were examined in the calves with respiratory and diarrheal disease. The result obtained in this investigation were as follows : 1. Among the 120 heads of calves that were purchased from market, 14 heads(22%) of calves were occurred respiratory disease, and 12 heads(20%) of calves are occurred diarrhea. Occurrence of respiratory disease was 14.5(4~20) days in average and diarrhea was 9.6(2-15) days after they had been purchased. 2. Serum protein fraction were analyzed by cellulose acetate electrophoresis. ${\beta}-globulin$, A/G ratio and ${\beta}_2-globulin$ values were decreased in the calves with respiratory disease. Especially, ${\beta}_2-globulin$ were significantly decreased. In calves with diarrhea, there was no change in ${\beta}-globulin$ values. ${\beta}_2-globulin$ values were higher than that of the normal and respiratory diseased calves. 3. ${\alpha}-globulin$ values were increased in both of calves with diarrhea and respiratory disease. This tendency was due to increase ${\alpha}_2-globulin$ values. 4. The $\gamma$-globulin value of calves with diarrhea was the lowest among the 3 groups. 5. The total protein values of normal calves were $7.0{\pm}1.1g/dl$ and that of respiratory and diarrheal diseased calves were $6.9{\pm}0.9g/dl$ and $6.6{\pm}0.8g/dl$, respectively. Total protein value of calves with diarrhea was lower than that of normal and respiratory diseased calves. Globulin value of calves with diarrhea was the lowest among them. The low value of total protein in diarrheal diseased calves was due to decrease globulin values. 6. The fibrinogen values of calves with respiratory disease ($643{\pm}189mg/dl$) were significantly higher than that of normal calves($533{\pm}135mg/dl$) and calves with diarrhea($572{\pm}188mg/dl$). The plasma : fib. ratio of respiratory diseased calves was $12.0{\pm}4.9$, normal calves was $13.8{\pm}3.5$ and diarrheal diseased calves was $12.8{\pm}4.6$. The ratio of the calves with respiratory disease was significantly decreased.

토끼에서 출생 후 고환간질세포의 발생에 관한 연구

태현진,박영재,강형섭,김남수,박상열,양홍현,안동춘,김인식,Tae, Hyun-Jin,Park, Young-Jae,Kang, Hyung-Sub,Kim, Nam-Soo,Park, Sang-Youel,Yang, Hong-Hyun,Ahn, Dong-Choon,Kim, In-Shik 대한수의학회 2005 大韓獸醫學會誌 Vol.45 No.3

Changes in the rabbit Leydig cell from birth to adulthood were studied in New Zealand white rabbits of 1, 7, 21, 35, 49, 70, 105, 147, 196, and 252 days (n = 8 rabbits per group) of age. The objectives of this study were to understand the fate of the fetal Leydig cells, to determine the changes in serum testosterone levels, and leutenizing hormone-stimulated testosterone production per testis in vitro, and to quantify adult Leydig cells by number and average volume with age. Testes of rabbits were fixed by whole body perfusion using a fixative containing 2.5% glutaraldehyde in cacodylate buffer, processed and embedded in Epon-araldite. Using $1{\mu}m$ sections stained with methylene blue-azure II, qualitative and quantitative (stereological) morphological studies were performed. Testosterone levels in the incubation medium of luteinizing hormone-stimulated (100 ng/ml) testosterone secretion per testis in vitro, and in serum were determined by radioimmunoassay. The average volume of a testis of 1-day-old rabbits was determined as $0.0073cm^3$ and the parameter increased linearly from birth to 252 days ($3.93cm^3$). The volume density of the seminiferous tubules increased with age from 33.76% at day 1 to 88.2% at day 252. The volume density of the interstitium represents 66.24% of the testicular parenchyma at day 1. This proportion progressively diminished during development to reach a value of 11.8% at day 252. The volume density of Leydig cells increased almost linearly from birth (0.001%) to 252 days (2.62%). Leydig cell mass per testis increases from 0.0012 mg to 0.25 mg between days 1 and 35, from 2.66 mg to 44.3 mg between days 49 and 105 and from 65.42 mg and 102.9 mg between days 147 and 252. The absolute numbers of adult Leydig cells per testis increased linearly from birth to 252 days. The average volume of adult Leydig cell on days 1, 7, 21 and 35 was not significantly different; a gradual and continued increase was observed thereafter, reaching a 3-fold increase at 196 and 252 days. Serum testosterone concentrations were not significantly different at day 1 compared days 7, 21, 35. Significant increases were observed at days 49 and 70. Values at days 70 and 105 and days 147, 196, and 252 were not significantly different. LH-stimulated testosterone production per testis in vitro was significantly different at day 1 compared days 7, 21, 35. Significant increases were observed at days 49 and 70. Hormonal values at days 105, 147, 196, and 252 were not significantly different. These data suggested Leydig cell developmental phase can be classified: a neonatal phase (1-7 days), a prepubertal phase (14-49 days) and an adult phase (70-252 days). Immature and mature adult Leydig cells, initially detected at days 7 and 49, respectively, and mature adult Leydig cells were abundant Leydig cell type according to the number and absolute volume per testis form day 49 onwards.

Gemcitabine의 세포사멸 기전 연구

설재원,이유진,강동원,강형섭,김남수,김인식,박상열,Seol, Jae-Won,Lee, You-Jin,Kang, Dong-Won,Kang, Hyung-Sub,Kim, Nam-Soo,Kim, In-Shik,Park, Sang-Youel 대한수의학회 2005 大韓獸醫學會誌 Vol.45 No.4

The nucleoside analogue gemcitabine (2', 2-difluorideoxycytide) is potential against a wide variety of solid tumors and considered to be one of the most active drugs in the treatment of non-small cell lung cancer (NSCLC). In this study, we investigated the signals of gemcitabine-induced apoptosis, especially in point of caspase pathway in A549. We exposed A549 cells to gemcitabine for dose/time dependent manner and the results showed that gemcitabine induced apoptotic cell death in a time/dose-dependent manner. We also treated to gemcitabine and Z-VAD-fmk as a pan-caspase inhibitor for 24 hours. Gemcitabine alone induced 35.3% cell death, and co-treatment with gemcitabine and Z-VAD-fmk induced 15.1% apoptotic cell death. Our results demonstrated that Z-VAD-fmk as a pan-caspase did not completely block the gemcitabine-induced apoptosis. Western blotting analysis showed that gemcitabine increased caspase-3, active caspase-8, p21 and p53 protein expressions in A549. Co-treatment with Z-VAD-fmk completely blocked caspase-3 and active caspase-8 protein expressions, but did not change the level of p21 and p53 protein expressions. Our data indicate that gemcitabine induced apoptosis through caspase-dependent and -independent pathways in A549.

산삼배양액 급여에 따른 육계의 생산성 및 질병 저항성 효과

설재원,박재홍,채준석,강형섭,류경선,강춘성,박상열,Seol, Jae-Won,Park, Jae-Hong,Chae, Joon-Seok,Kang, Hyung-Sub,Ryu, Kyeong-Seon,Kang, Chun-Seong,Park, Sang-Youel 대한수의학회 2010 大韓獸醫學會誌 Vol.50 No.2

The large amount of tissue culture medium (TCM), which contains some of the active secretory components of Korean wild ginseng (KWG; Panax ginseng) such as saponins, is usually discarded after harvest of KWG. The present study was aimed to investigate the efficacy of oral administration of the TCM-KWG on growth performance and diseases resistance in broiler chickens. A day old broiler chickens randomized in 6 groups (n = 60/groups) were administered orally with 0, 2, 4, 8, 16 and 32 mL/L TCMKWG through drinking water for 5 weeks and examined the change of weight gain, feed intake and blood components. Also, five weeks old broiler chickens (n = 15/groups) were challenged orally with Salmonella (S.) gallinarum and investigated the mortality in broiler chickens. An average weight gain and feed intake significantly didn't change in TCM-KWG administration groups as compared to control group. The concentration of calcium (Ca), phosphate (Pi) and potassium (K) in serum were increase by TCM-KWG administration in broiler chickens. We also found that oral administration of TCM-KWG through drinking water significantly reduced the mortality in broiler chickens experimentally infected with virulent S. gallinarum. The results of this study indicated that TCM-KWG administration may elevate the resistance on disease and improved the skeleton formation and body homeostasis of chickens, and TCM-KWG can be used as a cost-effective and environmentally alternative additives to control of the disease and growth.

산삼 배양액을 급여한 육계에서 근육의 프로테옴 분석

설재원,황인호,채준석,강형섭,류경선,강춘성,박상열,Seol, Jae-Won,Hwang, In Ho,Chae, Joon-Seok,Kang, Hyung-Sub,Ryu, Kyeong-Seon,Kang, Chun-Seong,Park, Sang-Youel 대한수의학회 2005 大韓獸醫學會誌 Vol.45 No.2

Proteomics is a useful approach to know protein expression, post-translational modification and protein function. We investigated the protein expression pattern and identity in chickens fed with the tissue culture medium waste after harvest of Korean wild ginseng (TCM-KWG) (Panax ginseng). Two groups (n=60/group) of day old broiler chickens were administered with 0 (control) and 0.8% (treatment) TCM-KWG through drinking water. After 5 weeks, we examined the protein expression pattern of fibularis longus and superficial pectoral muscle by Two-dimensional electrophoresis analysis. Interestingly, TCM-KWG treatment significantly increased five spot's density, and markedly reduced five spot's density in the muscles. We identified 10 proteins (desmin, myosin light chain 1, heat shock 25 kDa protein, collapsin response mediator protein-2A, alpha enolase, vimentin, actin alpha 1, my023 protein, pyruvate kinase and troponin T) by the matrix-assisted laser desorption ionization time of flight (MALDI-TOF).