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引用次数: 0
A useful model to study the effect of high sugar concentrations upon growth and enzymic activities of toad embryos and larvae 研究高糖浓度对蟾蜍胚和幼虫生长和酶活性影响的有效模型
Flavio Francini , Mariana Picasso , Oscar R Rebolledo , Alfredo Salibián , Juan José Gagliardino

The aim of this study was to develop an oviparous model suitable for studying the differential effects and mechanisms by which a high concentration of extracellular glucose and other sugars produce diabetes complications, particularly body growth retardation during development. Hence, we studied the experimental conditions necessary to obtain measurable effects of high sugar concentrations (5-mM glucose, mannitol, fructose and galactose) upon body growth and development of Bufo arenarum embryos and larvae, and upon the activity of aspartate aminotransferase (AST), γ-glutamyltransferase (GGT), and alkaline phosphatase (APP). Unfed animals kept in glucose showed lower body weight than controls at all stages, a condition only observed at stage 26 for animals kept in galactose and fructose. All animals reached the same stage of development regardless of the solution in which they were kept. Glucose and fructose significantly decreased the activity of all enzymes tested, while galactose only affected GGT activity. The model provides the first experimental evidence for the deleterious effect exerted in vivo by different sugars upon developing embryos and larvaes of Bufo arenarum. The results prove that this model might help to elucidate the effects and the pathogenic mechanisms of hyperglycemia upon growth and development of embryos exposed to environments with high sugar concentrations. It might also become a useful tool for testing the effectiveness of drugs designed to prevent the deleterious effect of such exposure.

本研究的目的是建立一个适合于研究高浓度细胞外葡萄糖和其他糖导致糖尿病并发症,特别是发育过程中身体生长迟缓的差异效应和机制的卵生模型。因此,我们研究了高糖浓度(5-mM葡萄糖、甘露醇、果糖和半乳糖)对砂蟾胚胎和幼虫身体生长发育以及对天冬氨酸转氨酶(AST)、γ-谷氨酰转移酶(GGT)和碱性磷酸酶(APP)活性的可测量影响所需的实验条件。葡萄糖喂养的未喂食动物在所有阶段的体重都低于对照组,这种情况只在第26阶段的半乳糖和果糖喂养的动物中观察到。所有的动物都达到了相同的发育阶段,不管它们被放在什么溶液中。葡萄糖和果糖显著降低了所有酶的活性,而半乳糖仅影响GGT活性。该模型首次提供了体内不同糖对斗牛鱼胚胎和幼虫的有害影响的实验证据。结果表明,该模型可能有助于阐明高血糖对高糖环境下胚胎生长发育的影响及其致病机制。它也可能成为一种有用的工具,用于测试旨在防止这种接触的有害影响的药物的有效性。
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引用次数: 1
Growth hormone inhibits growth hormone secretion from the rainbow trout pituitary in vitro 生长激素抑制虹鳟垂体生长激素的体外分泌
Thorleifur Ágústsson, Björn Thrandur Björnsson

Growth hormone (GH) secretion in salmonids and other fish is under the control of a number of hypothalamic factors, but negative feed-back regulation by circulating hormones can also be of importance for the regulation of GH secretion. Mammalian studies show that GH has a negative feed-back effect on its own secretion. In order to elucidate if GH levels present a direct ultra-short negative feedback loop at the pituitary level GH secretion was studied in intact pituitaries from 50 g fish in an in vitro perifusion system. Following an initial equilibrium period pituitaries were exposed to five increasing concentrations (1–1000 ng ml−1) of ovine GH (oGH) in 20-min steps, before being returned to a GH-free perifusion. Ovine GH caused a significant dose-dependant inhibition of GH secretion and it is concluded that GH can exert a direct negative feedback control on GH secretion at the pituitary level.

鲑鱼和其他鱼类的生长激素(GH)分泌受许多下丘脑因素的控制,但循环激素的负反馈调节也可能对GH分泌的调节起重要作用。哺乳动物研究表明,生长激素对其自身分泌有负反馈作用。为了阐明生长激素水平在垂体水平是否存在直接的超短负反馈回路,我们在体外灌注系统中研究了50 g鱼的完整垂体的生长激素分泌。在初始平衡期后,将垂体暴露于5种增加浓度(1 - 1000 ng ml - 1)的羊GH (oGH)中,分20分钟进行,然后返回无GH灌注。绵羊生长激素对生长激素分泌具有明显的剂量依赖性抑制作用,表明生长激素在垂体水平上对生长激素分泌具有直接的负反馈控制作用。
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引用次数: 29
Whole-body metabolism of the organophosphorus pesticide, fenthion, in goldfish, Carassius auratus 有机磷农药倍硫磷在鲫鱼体内的全身代谢
Shigeyuki Kitamura, Tomoko Kadota, Mayumi Yoshida, Norimasa Jinno, Shigeru Ohta

The in vivo metabolism of fenthion, an organophosphorus pesticide, and its sulfoxide (fenthion sulfoxide) was examined in goldfish (Carassius auratus). When goldfish were administered fenthion i.p. at a dose of 100 mg/kg, two metabolites were isolated from the tank water. They were identified as fenthion sulfoxide and fenthion oxon, in which >P=S of fenthion is transformed to >P=O, by comparing their mass and UV spectra, and their behavior in HPLC and TLC, with those of authentic standards. However, fenthion sulfone was not detected as a metabolite. The amounts of fenthion, fenthion sulfoxide and fenthion oxon excreted within 4 days were 2.7, 3.4 and 2.5% of the initial dose of fenthion, respectively. Unchanged fenthion was detected in the body of the fish to the extent of 42–50% of the dose after 10 days, but fenthion sulfoxide and fenthion oxon showed very low concentrations. When fenthion sulfoxide was administered to the fish, about 70% of the dose was excreted unchanged into the tank water within 24 h, but little of the reduced compound, fenthion, was found. In contrast, fenthion was detected at 2.1% of dose in the body of goldfish as a metabolite of fenthion sulfoxide. The fact that fenthion is metabolized to the toxic oxon form in fish presumably has environmental and health implication for its use as a pesticide.

研究了有机磷农药倍硫磷及其亚砜在鲫鱼体内的代谢情况。以100 mg/kg的剂量给金鱼喂食倍硫磷,从鱼缸水中分离出两种代谢物。通过与标准品的质谱、紫外光谱、HPLC和TLC行为比较,鉴定为倍硫磷亚砜和倍硫磷氧,其中倍硫磷的>P=S转化为>P=O。但代谢物未检出倍硫磷砜。4 d内的倍硫磷、倍硫磷亚砜和倍硫磷氧的排泄量分别为初始剂量的2.7、3.4和2.5%。10天后,在鱼体内检测到未变化的倍硫磷,其浓度为剂量的42-50%,但倍硫磷亚砜和倍硫磷氧的浓度极低。当给鱼注射倍硫磷亚砜时,约70%的剂量在24小时内原原本本地排泄到水箱水中,但很少发现被还原的化合物倍硫磷。相比之下,在金鱼体内检测到的倍硫磷是倍硫磷亚砜的代谢物,剂量为2.1%。倍硫磷在鱼类体内代谢为有毒的氧氧形式,这一事实可能对其作为杀虫剂的使用具有环境和健康影响。
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引用次数: 20
Metabolism of benzo(a)pyrene by duck liver microsomes 鸭肝微粒体对苯并(a)芘的代谢
Sangeet Honey , Patrick O’Keefe , Adam T. Drahushuk , James R. Olson , Subodh Kumar , Harish C. Sikka

The metabolism of benzo(a)pyrene [BP], a model carcinogenic PAH, by hepatic microsomes of two duck species, mallard (Anas platyrhynchos) and common merganser (Mergus merganser americanus) collected from chemically-contaminated and relatively non-contaminated areas was investigated. The rate of metabolism of BP by liver microsomes of common merganser and mallard collected from polluted areas (2650±310 and 2200±310 pmol/min per mg microsomal protein, respectively) was significantly higher than that obtained with liver microsomes of the two species collected from non-polluted areas (334±33 and 231±30 pmol/min per mg microsomal protein, respectively). The level of cytochrome P-450 1A1 was significantly higher in the liver microsomes of both duck species from the polluted areas as compared to the ducks from the non-polluted areas. The major BP metabolites, including BP-9, 10-diol, BP-4, 5-diol, BP-7, 8-diol, BP-1, 6-dione, BP-3, 6-dione, BP-6, 12-dione, 9-hydroxy-BP and 3-hydroxy-BP, formed by liver microsomes of both duck species from polluted and non-polluted areas, were qualitatively similar. However, the patterns of these metabolites were considerably different from each other. Liver microsomes of ducks from the polluted areas produced a higher proportion of benzo-ring dihydrodiols than the liver microsomes of ducks from the non-polluted areas, which converted a greater proportion of BP to BP-phenols. The predominant enantiomer of BP-7,8-diol formed by hepatic microsomes of the two duck species had an (−)R,R absolute stereochemistry. The data suggest that duck and rat liver microsomal enzymes have different regioselectivity but similar stereoselectivity in the metabolism of BP.

研究了从化学污染地区和相对非污染地区采集的绿头鸭(Anas platyrhynchos)和秋沙鸭(Mergus merganser americanus)两种鸭的肝脏微粒体对典型致癌性多环芳烃苯并(a)芘(BP)的代谢。污染地区沙鸭和绿头鸭肝脏微粒体对BP的代谢率(分别为2650±310和2200±310 pmol/min / mg微粒体蛋白)显著高于非污染地区沙鸭和绿头鸭肝脏微粒体对BP的代谢率(分别为334±33和231±30 pmol/min / mg微粒体蛋白)。污染地区鸭肝微粒体细胞色素P-450 - 1A1水平显著高于非污染地区鸭。污染地区和非污染地区鸭肝微粒体中BP的主要代谢产物BP-9、10-二醇、BP-4、5-二醇、BP-7、8-二醇、BP-1、6-二酮、BP-3、6-二酮、BP-6、12-二酮、9-羟基BP和3-羟基BP的质量相似。然而,这些代谢物的模式彼此有很大的不同。来自污染地区的鸭肝微粒体产生的苯并环二氢二醇的比例高于来自非污染地区的鸭肝微粒体,后者将BP转化为BP-酚的比例更高。两种鸭肝微粒体形成的bp -7,8-二醇的优势对映体具有(−)R,R绝对立体化学。结果表明,鸭和大鼠肝微粒体酶在BP代谢中具有不同的区域选择性,但具有相似的立体选择性。
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引用次数: 17
Inhibition of complement activation by water-soluble polysaccharides of some far-eastern brown seaweeds 一些远东褐藻水溶性多糖对补体活化的抑制作用
Tatiana N. Zvyagintseva , Natalia M. Shevchenko , Irina V. Nazarova , Andrey S. Scobun , Pavel A. Luk’yanov , Ludmila A. Elyakova

Fucoidans and laminarans from Laminaria cichorioides, Laminaria japonica, Fucus evanescens, laminaran from Laminaria gurjanovae, other β-d-glucans (translam, pustulan and zymosan) and λ-carrageenan from Chondrus armatus were used to study the effect of water-soluble polysaccharides from seaweeds on the alternative pathway of complement (APC). β-d-Glucans and fucoidans under study differed appreciably from each other by structural characteristics, and also by degree of purification. β-d-glucans, on ability to bind complement, ranked in a line according to a degree of their purification. Highly purified β-d-glucans under study did not reveal an ability to bind complement. The fucoidans were divided conventionally into three groups according to their action on APC. Highly sulfated α-l-fucan from L. cichorioides with the greatest activity toward APC and caused 50% inhibition of reaction of activation (RA) of APC in a concentration of 0.5–0.7 mg/ml. Opposite 50% of inhibition of lysis of erythrocytes by sulfated heterogeneous fucoidan from L.japonica was achieved with 20 mg/ml. All other fucoidans and λ-carrageenan have activity at 6–10 mg/ml concentration. Decreasing the sulfate content from 36% up to 9% in sample fucoidans under study was not reflected practically in the 50% inhibition concentration. Apparently, the degree of sulfating of fucoidans did not influence their action on APC. But the positive influence of fucose in structure of polysaccharide was obvious.

利用海带水溶性多糖对补体替代途径(APC)的影响,研究了海带水溶性多糖、日本海带水溶性多糖、海带水溶性多糖、海带水溶性多糖、海带水溶性多糖、海带水溶性多糖、海带水溶性多糖、海带水溶性多糖、海带水溶性多糖、海带水溶性多糖和海带水溶性多糖。所研究的β-d-葡聚糖和岩藻糖聚糖在结构特征和纯化程度上存在明显差异。β-d-葡聚糖结合补体的能力根据其纯化程度排列成一行。高纯度的β-d-葡聚糖在研究中没有显示出与补体结合的能力。岩藻多糖根据其对APC的作用通常分为三组。在0.5 ~ 0.7 mg/ml浓度下,对APC的活化反应(RA)有50%的抑制作用,对APC的活性最大。相反,20 mg/ml的硫酸酸化褐藻糖聚糖对红细胞裂解的抑制作用为50%。所有其他岩藻胶和λ-卡拉胶在6-10 mg/ml浓度下具有活性。在研究的岩藻胶样品中,硫酸盐含量从36%降低到9%,实际上并没有反映在50%的抑制浓度上。显然,岩藻胶的硫酸酸化程度不影响其对APC的作用。但焦点对多糖结构的积极影响是明显的。
{"title":"Inhibition of complement activation by water-soluble polysaccharides of some far-eastern brown seaweeds","authors":"Tatiana N. Zvyagintseva ,&nbsp;Natalia M. Shevchenko ,&nbsp;Irina V. Nazarova ,&nbsp;Andrey S. Scobun ,&nbsp;Pavel A. Luk’yanov ,&nbsp;Ludmila A. Elyakova","doi":"10.1016/S0742-8413(00)00114-6","DOIUrl":"10.1016/S0742-8413(00)00114-6","url":null,"abstract":"<div><p>Fucoidans and laminarans from <em>Laminaria cichorioides</em>, <em>Laminaria japonica,</em> <em>Fucus evanescens,</em> laminaran from <em>Laminaria gurjanovae</em>, other β-<span>d</span>-glucans (translam, pustulan and zymosan) and λ-carrageenan from <em>Chondrus armatus</em> were used to study the effect of water-soluble polysaccharides from seaweeds on the alternative pathway of complement (APC). β-<span>d</span>-Glucans and fucoidans under study differed appreciably from each other by structural characteristics, and also by degree of purification. β-<span>d</span>-glucans, on ability to bind complement, ranked in a line according to a degree of their purification. Highly purified β-<span>d</span>-glucans under study did not reveal an ability to bind complement. The fucoidans were divided conventionally into three groups according to their action on APC. Highly sulfated α-<span>l</span>-fucan from <em>L. cichorioides</em> with the greatest activity toward APC and caused 50% inhibition of reaction of activation (RA) of APC in a concentration of 0.5–0.7 mg/ml. Opposite 50% of inhibition of lysis of erythrocytes by sulfated heterogeneous fucoidan from <em>L.japonica</em> was achieved with 20 mg/ml. All other fucoidans and λ-carrageenan have activity at 6–10 mg/ml concentration. Decreasing the sulfate content from 36% up to 9% in sample fucoidans under study was not reflected practically in the 50% inhibition concentration. Apparently, the degree of sulfating of fucoidans did not influence their action on APC. But the positive influence of fucose in structure of polysaccharide was obvious.</p></div>","PeriodicalId":10586,"journal":{"name":"Comparative Biochemistry and Physiology Part C: Pharmacology, Toxicology and Endocrinology","volume":"126 3","pages":"Pages 209-215"},"PeriodicalIF":0.0,"publicationDate":"2000-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S0742-8413(00)00114-6","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21876699","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 43
Effect of three structurally related antimalarial drugs on liver microsomal components and lipid peroxidation in rats 三种结构相关抗疟药对大鼠肝微粒体成分和脂质过氧化的影响
E.Olatunde Farombi, Babatunde I Olowu, Godwin O Emerole

Changes in microsomal drug oxidizing enzymes, microsomal lipids, hepatic glutathione (GSH), glutathione S-transferase (GST) and malondialdehyde (MDA) formation following administration of rats with therapeutic doses of three structurally related antimalarial drugs, amodiaquine (AQ), mefloquine (MQ) and halofantrine (HF) were investigated. There was a significant decrease in the activities of aniline hydroxylase, p-nitroanisole O-demethylase and pentoxyresorufin O-dealkylase in AQ, MQ and HF treated rats. AQ elicited the greatest effect with 50, 37 and 67% reductions in the activities of aniline hydroxylase, p-nitroanisole O-demethylase and pentoxyresorufin O-dealkylase, respectively. All the drugs prolonged hexobarbital-sleeping time to varying extents. The three drugs increased significantly the cholesterol per phospholipid ratio. AQ, MQ and HF decreased significantly the GSH level, GST activity and increased the formation of MDA. The results indicate that the alterations in hepatic microsomal components and lipid peroxidation caused by the antimalarials are related to the structural differences in the compounds.

研究了大鼠给药后微粒体药物氧化酶、微粒体脂质、肝谷胱甘肽(GSH)、谷胱甘肽s -转移酶(GST)和丙二醛(MDA)形成的变化。AQ、MQ和HF处理大鼠的苯胺羟化酶、对硝基苯甲醚o -去甲基化酶和戊氧基间苯二酚o -脱烷基酶活性均显著降低。AQ对苯胺羟化酶、对硝基苯甲醚o -去甲基化酶和戊氧基间苯二酚o -脱烷基酶活性分别降低50%、37%和67%,影响最大。所有药物都不同程度地延长了六巴比妥睡眠时间。三种药物均显著提高胆固醇/磷脂比值。AQ、MQ和HF显著降低GSH水平、GST活性,增加MDA的形成。结果表明,抗疟药引起的肝微粒体成分和脂质过氧化的改变与化合物的结构差异有关。
{"title":"Effect of three structurally related antimalarial drugs on liver microsomal components and lipid peroxidation in rats","authors":"E.Olatunde Farombi,&nbsp;Babatunde I Olowu,&nbsp;Godwin O Emerole","doi":"10.1016/S0742-8413(00)00116-X","DOIUrl":"10.1016/S0742-8413(00)00116-X","url":null,"abstract":"<div><p>Changes in microsomal drug oxidizing enzymes, microsomal lipids, hepatic glutathione (GSH), glutathione <em>S</em>-transferase (GST) and malondialdehyde (MDA) formation following administration of rats with therapeutic doses of three structurally related antimalarial drugs, amodiaquine (AQ), mefloquine (MQ) and halofantrine (HF) were investigated. There was a significant decrease in the activities of aniline hydroxylase, <em>p</em>-nitroanisole <em>O</em>-demethylase and pentoxyresorufin <em>O</em>-dealkylase in AQ, MQ and HF treated rats. AQ elicited the greatest effect with 50, 37 and 67% reductions in the activities of aniline hydroxylase, <em>p</em>-nitroanisole <em>O</em>-demethylase and pentoxyresorufin <em>O</em>-dealkylase, respectively. All the drugs prolonged hexobarbital-sleeping time to varying extents. The three drugs increased significantly the cholesterol per phospholipid ratio. AQ, MQ and HF decreased significantly the GSH level, GST activity and increased the formation of MDA. The results indicate that the alterations in hepatic microsomal components and lipid peroxidation caused by the antimalarials are related to the structural differences in the compounds.</p></div>","PeriodicalId":10586,"journal":{"name":"Comparative Biochemistry and Physiology Part C: Pharmacology, Toxicology and Endocrinology","volume":"126 3","pages":"Pages 217-224"},"PeriodicalIF":0.0,"publicationDate":"2000-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S0742-8413(00)00116-X","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21876700","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 56
Accumulation of protoporphyrin-IX (PpIX) in leukemic cell lines following induction by 5-aminolevulinic acid (ALA) 5-氨基乙酰丙酸诱导白血病细胞系中原卟啉- ix (PpIX)的积累
Jitka Bartošová, Zbyněk Hrkal

We investigated the amounts of protoporphyrin IX (PpIX) accumulated in noninduced cells and following 5-aminolevulinic acid (ALA)-induction. Following ALA administration PpIX increased in all leukemic cell lines under investigation (HEL 26-fold, HL60 6-fold, Jurkat 3-fold, ML2 2-fold) but not in lymphocytes. Compared to other cell lines studied, HEL cells showed the lowest basal level of PpIX and the largest relative increase in PpIX. Despite a high increase following ALA treatment, the PpIX level in HEL cells is almost as low as in lymphocytes. It is in agreement with their relatively low sensitivities of ALA-induced photodynamic therapy (ALA-PDT) shown previously [(Grebeňová, D., Cajthamlová, H., Bartošová, J., Marinov, J., Klamová, H., Fuchs, O., Hrkal, Z., 1998. Selective destruction of leukemic cells by photo-activation of 5-aminolevulinic acid –induced protoporphyrin IX. J. Photochem. Photobiol. B: Biol. 47, 74–81)]. The ferrochelatase activities in the individual cell lines are in good inverse correlation with PpIX amounts accumulated in the ALA-induced cells, but not with the relative increase (ratio) of PpIX levels from basal to ALA-induced ones. This is most apparent in HEL cells and lymphocytes. There is probably different regulation of heme biosynthesis in erythroid cells, which are therefore not suitable for the studies of ALA-PDT mechanism. PpIX was accumulated more extensively in absence of fetal calf serum than in its presence. The amounts of PpIX accumulated in cells decreased exponentially with increasing fetal calf serum concentration.

我们研究了非诱导细胞和5-氨基乙酰丙酸(ALA)诱导后原卟啉IX (PpIX)的积累量。在接受ALA治疗后,PpIX在所有白血病细胞系中均增加(HEL 26倍,HL60 6倍,Jurkat 3倍,ML2 2倍),但在淋巴细胞中没有增加。与研究的其他细胞系相比,HEL细胞PpIX的基础水平最低,PpIX的相对增幅最大。尽管ALA治疗后PpIX水平高,但HEL细胞中的PpIX水平几乎与淋巴细胞一样低。这与先前显示的ala诱导的光动力疗法(ALA-PDT)相对较低的灵敏度一致[(Grebeňová, D., cajthamlov, H., Bartošová, J., Marinov, J., klamov, H., Fuchs, O., Hrkal, Z., 1998.]。5-氨基乙酰丙酸诱导的原卟啉IX光激活对白血病细胞的选择性破坏。j . Photochem。Photobiol。[B].生物学报。47,74-81。单个细胞系中铁螯合酶活性与ala诱导细胞中PpIX的积累量呈良好的负相关,但与PpIX水平从基础到ala诱导的相对增加(比率)无关。这在HEL细胞和淋巴细胞中最为明显。红细胞血红素生物合成可能有不同的调控机制,因此不适合ALA-PDT机制的研究。在没有胎牛血清的情况下,PpIX的积累比有胎牛血清的情况下更广泛。随着胎牛血清浓度的增加,细胞中PpIX的积累量呈指数下降。
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引用次数: 27
The house musk shrew (Suncus murinus): a unique animal with extremely low level of expression of mRNAs for CYP3A and flavin-containing monooxygenase 麝香鼩(Suncus murinus):一种独特的动物,其CYP3A和含黄素单加氧酶mrna的表达水平极低
Taisei Mushiroda , Tsuyoshi Yokoi , Kunio Ito , Ken-ichi Nunoya , Tetsuya Nakagawa , Megumi Kubota , Eiji Takahara , Osamu Nagata , Hideo Kato , Tetsuya Kamataki

Expression of drug-metabolizing enzymes including cytochrome P450 (CYP) and flavin-containing monooxygenase (FMO) in various tissues of Suncus murinus (Suncus) were examined. Northern blot analysis showed that mRNAs hybridizable with cDNAs for rat CYP1A2, human CYP2D6, rat CYP2B1, human CYP2C8, human CYP2136, rat CYP2E1, human CYP3A4 and rat CYP4A1 were expressed in various tissues from Suncus. The mRNA level of CYP2A in the Suncus lung was very high. Furthermore, it was found that the level of CYP2A mRNA in the Suncus lung was higher compared to the Suncus liver. The expression level of mRNA hybridizable with cDNA for human CYP3A4 was very low. The presence of CYP3A gene in Suncus was proven by the induction of the CYP with dexamethasone. Very low expression levels of mRNAs hybridizable with cDNAs for rat FMO1, rat FMO2, rat FMO3 and rat FMO5 were also seen in Suncus liver. No apparent hybridization band appeared when human FMO4 cDNA was used as a probe. The hepatic expression of mRNAs hybridizable with cDNAs for UDP-glucuronosyltransferase 1*6, aryl sulfotransferase, glutathione S-transferase 1, carboxyesterase and microsomal epoxide hydrolase in the Suncus were observed. These results indicate that the Suncus is a unique animal species in that mRNAs for CYP3A and FMO are expressed at very low levels.

研究了鼠子(Suncus murinus)各组织中细胞色素P450 (CYP)和含黄素单加氧酶(FMO)等药物代谢酶的表达。Northern blot分析显示,与大鼠CYP1A2、人CYP2D6、大鼠CYP2B1、人CYP2C8、人CYP2136、大鼠CYP2E1、人CYP3A4和大鼠CYP4A1的cdna杂交的mrna在Suncus各组织中均有表达。Suncus肺组织中CYP2A mRNA水平非常高。此外,我们还发现Suncus肺中CYP2A mRNA的水平高于Suncus肝。人CYP3A4可与cDNA杂交的mRNA表达量很低。通过地塞米松诱导Suncus中CYP3A基因的存在证实了CYP3A基因的存在。大鼠FMO1、FMO2、FMO3和FMO5的mrna与cdna的杂交表达水平也很低。以人FMO4 cDNA为探针,未发现明显的杂交带。观察了苏家鱼肝脏中udp -葡萄糖醛基转移酶1*6、芳基磺基转移酶、谷胱甘肽s -转移酶1、羧化酶和微粒体环氧化物水解酶的mrna表达。这些结果表明,Suncus是一种独特的动物物种,CYP3A和FMO的mrna表达水平非常低。
{"title":"The house musk shrew (Suncus murinus): a unique animal with extremely low level of expression of mRNAs for CYP3A and flavin-containing monooxygenase","authors":"Taisei Mushiroda ,&nbsp;Tsuyoshi Yokoi ,&nbsp;Kunio Ito ,&nbsp;Ken-ichi Nunoya ,&nbsp;Tetsuya Nakagawa ,&nbsp;Megumi Kubota ,&nbsp;Eiji Takahara ,&nbsp;Osamu Nagata ,&nbsp;Hideo Kato ,&nbsp;Tetsuya Kamataki","doi":"10.1016/S0742-8413(00)00113-4","DOIUrl":"10.1016/S0742-8413(00)00113-4","url":null,"abstract":"<div><p>Expression of drug-metabolizing enzymes including cytochrome P450 (CYP) and flavin-containing monooxygenase (FMO) in various tissues of <em>Suncus murinus</em> (<em>Suncus</em>) were examined. Northern blot analysis showed that mRNAs hybridizable with cDNAs for rat CYP1A2, human CYP2D6, rat CYP2B1, human CYP2C8, human CYP2136, rat CYP2E1, human CYP3A4 and rat CYP4A1 were expressed in various tissues from <em>Suncus.</em> The mRNA level of CYP2A in the Suncus lung was very high. Furthermore, it was found that the level of CYP2A mRNA in the Suncus lung was higher compared to the <em>Suncus</em> liver. The expression level of mRNA hybridizable with cDNA for human CYP3A4 was very low. The presence of CYP3A gene in <em>Suncus</em> was proven by the induction of the CYP with dexamethasone. Very low expression levels of mRNAs hybridizable with cDNAs for rat FMO1, rat FMO2, rat FMO3 and rat FMO5 were also seen in <em>Suncus</em> liver. No apparent hybridization band appeared when human FMO4 cDNA was used as a probe. The hepatic expression of mRNAs hybridizable with cDNAs for UDP-glucuronosyltransferase 1<sup>*</sup>6, aryl sulfotransferase, glutathione <em>S</em>-transferase <em>1</em>, carboxyesterase and microsomal epoxide hydrolase in the <em>Suncus</em> were observed. These results indicate that the <em>Suncus</em> is a unique animal species in that mRNAs for CYP3A and FMO are expressed at very low levels.</p></div>","PeriodicalId":10586,"journal":{"name":"Comparative Biochemistry and Physiology Part C: Pharmacology, Toxicology and Endocrinology","volume":"126 3","pages":"Pages 225-234"},"PeriodicalIF":0.0,"publicationDate":"2000-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S0742-8413(00)00113-4","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21876701","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 4
Further immunochemical and biocatalytic characterization of CYP1A1 from feral leaping mullet liver (Liza saliens) microsomes 野生跳乌鱼肝微粒体CYP1A1的进一步免疫化学和生物催化特性研究
Alaattin Şen , Emel Arinç

CYP1A is known to play important roles in the metabolism, detoxification and bioactivation of carcinogens and other xenobiotics in animals including fish. In our laboratory, CYP1A1 was obtained in a highly purified form with a specific content of 15–17 nmol P450 per mg protein from liver microsomes of feral fish, leaping mullet (Liza saliens). Purified mullet CYP1A1 showed a very high substrate specificities for 7-ethoxyresorufin and 7-methoxyresorufin in a reconstituted system containing purified fish P450 reductase and lipid. In addition, effects of each individual components of the reconstituted system, i.e., CYP1A1 and P450 reductase on 7-methoxyresorufin O-demethylase (MROD) activity were studied. 7-ethoxyresorufin O-deethylase (EROD) activity was strongly inhibited by α-naphthoflavone (ANF). At 0.5 and 2.5 μM, ANF inhibited EROD activity by 90 and 98%, respectively. Mullet CYP1A1 did not catalyze monooxygenations of other substrates such as aniline, ethylmorphine, N-nitrosodimethylamine and p-nitrophenol. Antibodies produced against CYP1A1 orthologues in fish such as trout and scup showed strong cross-reactivity with the purified mullet CYP1A1. In addition, anti-L. saliens liver CYP1A1 produced in our laboratory inhibited both the EROD and MROD activities catalyzed by L. saliens liver microsomes but stronger inhibition was observed with EROD activity. On the other hand, anti-mullet CYP1A1 antibodies showed very weak cross-reactivity with two proteins (presumably CYP1A1 and CYP1A2) in 3MC-treated rat liver microsomes. Moreover, 3MC-treated rat liver microsomal EROD activity was weakly inhibited by the anti-L. saliens liver CYP1A1. These results strongly suggested that the purified mullet CYP1A1 is structurally, functionally and immunochemically similar to the CYP1A1 homologues purified from other teleost species but functionally and immunochemically distinct from mammalian CYP1A1.

CYP1A在包括鱼类在内的动物体内的致癌物和其他外源物的代谢、解毒和生物活化中发挥重要作用。在我们的实验室中,CYP1A1以高度纯化的形式从野生鱼跳跃鲻(Liza saliens)的肝微粒体中获得,其特定含量为每毫克蛋白质15-17 nmol P450。在含有纯化鱼P450还原酶和脂质的重组体系中,纯化的鲻鱼CYP1A1对7-乙氧基间苯二酚和7-甲氧基间苯二酚具有非常高的底物特异性。此外,还研究了重组体系中各组分CYP1A1和P450还原酶对7-甲氧基间苯二酚o -去甲基化酶(MROD)活性的影响。α-萘黄酮(ANF)明显抑制7-乙氧基间苯二酚o -去乙基酶(EROD)活性。在0.5 μM和2.5 μM下,ANF分别抑制了90%和98%的EROD活性。Mullet CYP1A1不催化其他底物如苯胺、乙基吗啡、n -亚硝基二甲胺和对硝基苯酚的单氧作用。在鳟鱼和鲫鱼中产生的抗CYP1A1同源物的抗体与纯化的鲻鱼CYP1A1具有很强的交叉反应性。此外,anti-L。本实验室产生的saliens肝脏CYP1A1对L. saliens肝微粒体催化的EROD和MROD活性均有抑制作用,但对EROD活性的抑制作用更强。另一方面,抗鲻鱼CYP1A1抗体与3mc处理的大鼠肝微粒体中的两种蛋白(可能是CYP1A1和CYP1A2)表现出非常弱的交叉反应性。此外,3mc处理的大鼠肝微粒体EROD活性被抗- l弱抑制。肝脏CYP1A1。这些结果强烈表明,纯化的乌鱼CYP1A1在结构、功能和免疫化学上与从其他硬骨鱼物种纯化的CYP1A1同源物相似,但在功能和免疫化学上与哺乳动物的CYP1A1不同。
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引用次数: 17
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Comparative Biochemistry and Physiology Part C: Pharmacology, Toxicology and Endocrinology
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