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MATERIALS AND METHODS Female ICR mice (Harlan, Houston, Tex.) weighing 18-23 g were ran- domly divided into 3 groups of 20 and given powdered feed (Purina Lab Chow 5002) supplemented with 0, 0.125, and 0.5% EQ HCI, respectively. Mice were killed by cervical dislocation, 4/group, after 2, 4, 6, and 10 wk of feeding; the liver, kidney, lung, and brain tissues were rapidly excised and weighed. Approximately 200 mg of each tissue sample with the ex- ception of the lung tissue was homogenized in 10 volumes (w/v) of ice- cold acetonitrile-water (7 : 3, v/v) with a Polytron (Brinckmann), then cen- trifuged for 10 min at 5000 x g. The clear supernatant was transferred into a screw-capped glass vial, centrifuged, then stored in a freezer (-20°C) for 2-3 h or until the two layers separated. Due to the small size of the lung tissue, most of it was homogenized in 20 volumes of solvent. The clear upper layers, approximately 60% of the total volume, contained most (>90%) of the EQ; therefore, only the upper layers were analyzed by HPLC detected by the fluoromonitor. In order to avoid possible diur- nal fluctuations, mice were killed at 8:00-10:00. Approximately 200 mg of another liver sample from each mouse was homogenized in 10 volumes (w/v) of ice-cold 0.25 M perchloric acid and centrifuged for 10 min at 5000 x g for the determination of GSH level. Three groups of mice, 4/group, were killed after 14 wk of feeding, 1/group/d from each feeding group, and the mitochondria were isolated from the liver, saving 2 samples of approximately 200 mg each to deter- mine the EQ residue and GSH levels. The liver mitochondria were iso- lated in 2.5 M sucrose buffer by differential centrifugaron Gohnson and Lardy, 1967) and the protein concentration was determined by the biuret method (Gornall et al., 1949). |
至尊木蟲 (職業(yè)作家)
木蟲 (正式寫手)
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雌性ICR鼠(18-23g)隨機分成三組,每組20只并正常喂養(yǎng),飼料中分別添加0,0.125和0.5%的EQ HCI,在第2,4,6,10周的時候分別取每組中的四只老鼠斷頸處死,迅速取出肝,腎,肺,腦組織并稱重。大約200mg的組織(除去肺組織)懸浮于10倍體積預(yù)冷的乙腈-水(7:3體積比),然后5000g離心10min,上清液轉(zhuǎn)移到有蓋子的玻璃小瓶中,儲存在-20°2-3h或者等到兩相分離。由于肺組織比較小,一般懸浮于20倍體積的溶劑,清透的上清液大約占總體積的60%左右,包括EQ(超過90%),因此,只有上相才有高效液相分析,為了避免日常的波動,老鼠仔8點到十點之間處死。大約200mg肝組織懸浮于10倍體積預(yù)冷的0.25M過氯酸,5000g離心10min以達(dá)到檢測谷胱甘肽水平。 第14周處死的老鼠,每組中取一只從肝組織分離線粒體,大約200mg來檢測EQ和GSH水平,肝線粒體用2.5M蔗糖不同離心力來分離,用雙縮脲法檢測蛋白濃度。(太老的文獻(xiàn)了) 大概是這樣吧! |

榮譽版主 (文壇精英)
| 雌性ICR小鼠(哈倫,休斯敦,得克薩斯州),體重18-23克然進(jìn)行隨機分為3組,每組20和粉狀飼料(普瑞納實驗室周5002)輔以0,0.125,0.5%EQ人機交互,分別。頸椎脫位,4/group,后2,4,6,和10周處死小鼠,喂養(yǎng);肝,腎,肺,腦組織迅速切除并稱重。大約200毫克的每個組織樣本前肺組織勻漿10卷(W / V)冰ception冷乙腈 - 水(7:3,V / V),然后與寶創(chuàng)(Brinckmann)中心trifuged為5000 XĞ10分鐘。清澈的上清液轉(zhuǎn)移到一個螺絲帽的玻璃瓶,離心,然后存放在冰箱(-20℃)為2-3小時,或直至兩層分離。由于規(guī)模小肺組織,其中大部分被勻漿溶劑,在20卷。明確的上層,總量約60%,載大部分的EQ(> 90%),因此,只有上層進(jìn)行了分析高效液相色譜法檢測由fluoromonitor。為了避免可能diur信號波動,處死小鼠在8:00-10:00。約200毫克。另外從各組小鼠肝勻漿樣品10卷冰冷的0.25 M過氯酸(W / V),離心10分鐘G為5000 X GSH水平的決心。 三組小鼠,4/group,喂養(yǎng)14周后處死,1/group/d每次喂食組,線粒體分離從肝,節(jié)省約200毫克,每2個樣品阻止挖掘情商殘留和GSH水平。肝線粒體是ISO-lated差centrifugaron Gohnson和2.5 M蔗糖緩沖拉迪,1967年)和蛋白質(zhì)濃度的縮二脲的測定方法(Gornall等,1949)。撤消修改Cíxìng ICR xiǎo shǔ (hā lún, xiūsīdūn, dekèsàsī zhōu), tǐzhòng 18-23 kè rán Jìnxíng suíjī fēn wéi 3 zǔ, měi zǔ 20 hé fěn zhuàng sìliào (pǔ ruì nà shíyàn shì Zhōu 5002) fǔ yǐ 0,0.125, 0.5% EQ rén jī jiāohù, fēnbié. Jǐngchuí tuōwèi, 4/group, hòu 2,4,6, hé 10 zhōu chǔsǐ xiǎo shǔ, Wèiyǎng; gān, shèn, fèi, nǎo zǔzhī xùnsù qiēchú Bìng chēng zhòng. Dàyuē 200 háokè de měi gè zǔzhī yàngběn qián Fèi zǔzhī yún jiāng 10 juǎn (W / V) bīng ception Lěng yǐjīng - shuǐ (7: 3, V/ V), ránhòu yǔ bǎo chuàng (Brinckmann) zhōngxīn Trifuged wèi 5000 X Ğ 10 fēnzhōng. Qīngchè de shàng qīng yè zhuǎnyí Dào yīgè luósī mào de bōlí píng, líxīn, ránhòu cúnfàng zài bīngxiāng ( -20℃) Wèi 2-3 xiǎoshí, huò zhízhì liǎng céng fēnlí. Yóuyú guīmó xiǎo Fèi zǔzhī, qízhōng dà bùfèn bèi yún jiāng róngjì, zài 20 juǎn. Míngquè de shàngcéng, zǒng liàng yuē 60% , zài Dà bùfèn de EQ (> 90% ), yīncǐ, zhǐyǒu shàngcéng jìnxíngle fēnxī Gāoxiào yè xiāng sèpǔ fǎ jiǎncè yóu fluoromonitor. Wèile bìmiǎn kěnéng diur Xìnhào bōdòng, chǔsǐ xiǎo shǔ zài 8:00-10:00. Yuē 200 háokè. Lìngwài cóng gè zǔ xiǎo shǔ gān yún jiāng yàngpǐn 10 juǎn Bīnglěng de 0.25 Mguò lǜ suān (W / V), líxīn 10 fēnzhōng G wèi 5000 X GSH shuǐpíng de juéxīn. Sān zǔ xiǎo shǔ, 4/group, wèiyǎng 14 zhōu hòu chǔsǐ, 1/Group/d měi cì wèishí zǔ, xiànlìtǐ fēnlí Cóng gān, jiéshěng yuē 200 háokè, měi 2 gè yàngpǐn zǔzhǐ Wājué qíngshāng cánliú hé GSH shuǐpíng. Gān xiànlìtǐ shì ISO- Lated chà centrifugaron Gohnson hé 2.5 M zhètáng huǎnchōng Lā dí,1967 nián) hé dànbáizhí nóngdù de suō èr niào de cèdìng Fāngfǎ (Gornall děng, 1949). |

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