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Fusarium poae와 Fusarium sporotrichioides간의 원형질체 융합
하경란,장성렬,민병례 한국미생물학회 1991 미생물학회지 Vol.29 No.2
In order to develop the protoplast fusion method of the strains of Fusarium, the interspecific protoplast fusion was attempted between Fusarium poae and F. sporotrichioides. Various auxotrophic mutants were isolated by the treatment of N-Methyl-N'-Nitro-N-Nitrosoguanidine. The optimal conditions for the formation and regeneration of protoplasts were examined and the characteristics of a fusant were studied. As a results, protoplasts were readily obtained from 18 hours cultured mycelia by the treatment of driselase for 3 hours and 0.6 M KCl as a best osmotic stabilizer at pH 6.0 for the formation of protoplast. Sucrose was the most suitable for the regeneration. Polyetylene glycol (M.W. 8,000) in $CaCl_{2}$-glycine solution was used to induce the protoplast fusion. The interspecific fusion frequency between protoplasts among the auxotrophic mutants of the two strains ranged from $2.7*10^{-2}$ to $5.7*10^{-3}$ . DNA content and cellulase activity were rather increased in the interspecific fusant. The lag phase of growth curve was slightly elongated in the fusant.
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Cisplatin이 흰쥐 신장에서의 hydroxyl radical 생성과 지질 과산화에 미치는 영향
서대규,신인철,서경용,고현철,하경란 한양대학교 의과대학 1994 한양의대 학술지 Vol.14 No.1
Cisplatin(cis-diamminedichloroplatinum Ⅱ) is a chemotherapeutic agent which is important in the treatment of several human cancers. The major limitation in the use of cisplatin is nephrotoxicity which may occur acutely or after several courses of treatment. THe mechanism of cisplatin nephrotoxicity is unknown. For the determination of the role of free radical in cisplatin-induced nephrotoxicity, we examined the effects of cisplatin on the activities of catalase and content of malondialdehyde(MDA) in rats. The Wistar rats(n=40), weighing about 200gm, were injected intraperitonealy with a single dose of cisplatin(6mg/kg). The rats were decapitated and the left kidney were perfused with an ice cold phosphate buffered saline(pH 7.3). The renal cortex was separated mudullar from the kidney and homogenized in a 10mM potassium phosphate buffer(pH 7.3) and then the cell membranes were disrupted by sonication. THe catalase activity was determined by the method of Cohen which involves titration of KMnO₄and the MDA contents were determined by the method of thiobarbituric acid assay. The catalase activity and the MDA contents were expressed on a mg protein. The results obtained were as follows. 1.Catalase activity(k/mg protein) In control group(n=5), the catalase acivity(means±S.D.) resulted in 43.80 4.26 and the catalase activity of group treated with cisplatin(n=15) was increased 10.1% at the 1st day but decreased 60.4 and 39.8%(p<0.01) respectively on the 2nd day and the 3rd day after the injection as compared with the catalase activity of control(p<0.01). 2.Malondialdehyde(MDA) content(nmol/mg protein) In control group(n=5), MDA content(means±S.D.) resulted in 2.11±0.35 and significantly the content of groups treated with cisplatin("n=15) increased 46.4(p<0.05), 20.9, 35.1%(p<0.01) respectively on the 1st, the 2nd, the 3rd day after the injection as compared with MDA content of control. These results suggest that hydroxyl radicals are involved in pathogenesis of cisplatin-induced nephrotoxicity.
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흰쥐에서의 Ethanol 유발 간독성에 대한 Biphenyldimethyl dicarboxylate(DDB)의 보호효과
신인철,하경란,정낙은,고현철 한양대학교 의과대학 1995 한양의대 학술지 Vol.15 No.1
In an attempt to define the effects of Biphenyldimethyl dicarboxylate(DDB) on the lipid peroxidation and oxygen free radical scavenging enzymes activities in ethanol-induced hepatotoxic rats, we studied malondialdehyde(MDA) level and the activities of catalase and superoxide dismutase(SOD) in liver of the rats at 24hr after the injection of ethanol. Sprague-Dawley albino rats weighing 250 to 280gm were injected intraperitoneally with ethanol(2.5gm/kg) only and ethanol plus DDB. DDB(200mg/kg/day, P.O.) is administered for 4days prior to 3days from the injection of ethanol. The result obtained can be summarized as follows : 1. The group treated with ethanol showed significantly higher MDA level and lower catalase and SOD activities at 24hr after the injection as compared with that of control group. 2. The group treated with ethanol plus DDB showed significantly lower MDA level and higher catalase and SOD activities at 24hr after the injection as compared with that of ethanol group. These results suggest that the excessive oxygen free radicals resulting from the depression of the activities of catalase and superoxide dismutase is an important determinant in pathogenesis of ethanol-induced hepatotoxicity and DDB has antioxidant effects.
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염화 제2수은이 흰쥐 신장에서의 지질 과산화와 Catalase 활성도에 미치는 영향
이숭구,하경란,고현철,신인철,서대규 한양대학교 의과대학 1994 한양의대 학술지 Vol.14 No.2
In an attempt to define the early biochemical determinants that participate in the pathogenesis of mercuric chloride-induced nephrotoxicity, especially focusing on oxygen free radicals, we studies malondialdehyde(MDA) level and catalase activity in renal cortex of the rats at 24, 48 and 72 hr after the injection of mercuric chloride. Wistar albino rats weighing 180 to 220gm were injected subcutaneously with mercuric chloride(HgCl₂, 2mg/kg). The result obtained can be summarized as follows: 1.The group treated with mercuric chloride showed significantly lower MDA level at 24, 48 and 72hr after the infection as compared to that of control group. 2.The group treated with mercuric chloride showed significantly higher catalase activity at 24hr and lower catalase activity at 72hr after the infection as compared to that of control group. These results suggest that the excessive oxygen free radicals resulting from the depression of catalase activity is an important determinant in pathogenesis of mercuric chloride-induced nephrotoxicity.
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흰쥐에서의 Puromycin Aminonucleoside-유발 단백뇨에 대한 선택적 Thromboxane A₂수용체길항제, KT2-962의 효과
서대규,신인철,고현철,하경란,강주섭 한양대학교 의과대학 1994 한양의대 학술지 Vol.14 No.1
The administration of puromycin aminonucleoside(PAN) to rats caused to nephrotic syndrome which characterized ascites, proteinurisa, hypoalbuminemia, and hyperlipidemia similar to those observed in human minimal change disease. Recently, several studies indicate that renal endogenous thromboxane(Tx) A₂may have an important role in pathophysiology of various renal disease. In this sutdy, we hafve examined the protective effct of a selective TxA₂receptor antagonist, KT2-962(KT2) on PAN-induced proteinuria in rats. Thus, male Wistar rats were given either daily subsutaneous injection of PAN, 20mg/kg, for 10 consecutive days from 3 days before to 7 days with PAN treatment. Urine was collectd, and body weight was measured in interval of 2 days during 2 weeks and urinary N-acetyl-β-Dglucosaminidase(NAG) activity as an index of renal tubular cell damage and urine protein were measured. In addition to measuring BUN, serum creatinine and creatinine clearance were measured to assess the degree of renal functional damage in 14th day. The results(Means SE) otained can be summarized as follows: 1)Body weight(gm) was progressively increased and gained about 46.4gm and 39.2gm on 2 weeks of treatment in the control and KT2 groups respectively. In constrast, there was weight loss about 27.4gm in the PAN group. But, it was increased about 23.2 gm in KT2+PAN grou and means that KT2 has significantly(p<0.05) suppressed weight loss by PAN. 2)Urine flow (ml/24 hours) was slightly increased in both control and KT2 groups during 2 weeks. But, it was significantly(p<0.05) increased after 7th groups during 2 weeks. But, it was significantly(p<0.05) increased after 7th day. But, concurrent administration of KT2 significantly(P<0.05) suppressed PAN-induced polyuria in KT2+PAN group. 3)Urinary protein(mg/24 hours0 was slightly increased in both control and KT2 groups. But, it was progressively increased and reached at the maximal level, 3.2 folds of initial level to 11th day and thereafter slightly reduced proteinuria to 14th day in the PAN group. In contrast, KT2 cotreatment with PAN was significantly(P<0.05) suppressed PAN-induced proteinuria in the KT2+PAN group. 4)Urinary NAG activity was markedly increased and reached to maximal level, 122.03 18.53 U/mg of urine creatinine, 12.7 folds of initial by day 9 and thenafter progressively decreased to 5.4 folds of initial level by day 14 in the PAN group. But, when KT2 was administered with PAN, it was significantly depressed its increment to day 13. But, it was reached to maximal level, 99.05 42.55, 12.7 folds of inital level much than PAN group. This result indicated that KT2 had a partial preventive effect on PAn-induced renal tubular cell damage. 5)The BUN and serum creatinine level(mg/dl) were significantly(p<0.05) increased from initial level, 18.48 1.28 and 0.50 0.03 to 118.42 41.34 and 1.66 0.27 respectively, and creatinine clearance(ml/min) was significantly(P<0.05) decreased from initial level, 0.44 0.02 to 0.28 0.07 by day 14 by PAN treatment. But, when PAN was given together with KT2, the increment of BUN and serum creatinine except for creatinine clearance were significantly(P<0.05) inhibited in the KT2+PAN group. Based on all these results obtained in this study, it is concluded that the coadministration of KT2-962 with PAN can be ingibited protein excretion in urine and suggested that endogenous TxA₂would take part in PAN-induced proteinuria in rats.
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송호연(Ho-Yeon Song),하경란(Kyung-Ran Ha),고현철(Hyun-Chul Koh),신인철(In-Chul Shin),서대규(Tae-Kyu Suh) 대한약리학회 1994 대한약리학잡지 Vol.30 No.2
In an attempt to define the effects of Biphenyldimethyl dicarboxylate(DDB) on the lipid peroxidation, oxygen free radical scavenging enzymes activities and hepatic functions in ethanol-induced hepatotoxic rats, we studies malondialdehyde(MDA) level and the activities of catalse, superoxide dismutase(SOD), glutamic-oxaloacetic transaminase(GOT) and glutamic-pyruvic transaminase(GPT) in liver of the rats at 24, 48 and 72 hr after the injection of ethanol and DDB. Sprague-Dalwey albino rats weighing 250 to 280gm were injected intraperitoneally with ethanol(2.5 gm/kg ) only and ethanol plus DDB(300mg/kg ). The result obtained can be summarized as follows : 1) The group treated with ethanol showed significantly higher MDA level and lower catalase and SOD activities at 24, 48 and 72hr after the injection as compared with that of control group. 2) The group treated with ethanol showed significantly higher GOT and GPT activities at 24, 48 and 72hr after the injection as compared with that of control group. 3) The group treated with ethanol plus DDB showed significantly lower MDA level and higher catalase and SOD activities at 24, 48 and 72 hr after the injection as compared with that of ethanol group. 4) The group treated with ethanol plus DDB showed significantly lower GOT and GPT activities at 24, 48 and 72 hr after the injection as compared with that of ethanol group. These results suggest that the excessive oxygen free radicals resulting from the depression of the activities of catalase and superoxide dismutase is an important determinant in pathogenesis of ethanol-induced hepatotoxicity and DDB has antioxidant effects.
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