Metabolic factors affecting protein synthesis by lung in vitro.
Metabolic factors affecting protein synthesis by lung in vitro.
复制标题
影响肺体外蛋白质合成的代谢因素。
DOI：
10.1152/jappl.1971.30.1.1
复制
发表时间：
1971
影响因子：
3.3
通讯作者：
H. Steinkamp
中科院分区：
文献类型：
--
作者：
D. Massaro;M. Simon;H. Steinkamp
肺切片的材料与方法。研究人员给体重约2公斤的新西兰白化雄性兔子随意喂食和饮水，并将其击晕，然后将肺、心脏和附着结构从胸部整体切除。肺外表面冷冲洗0.85，在0.5-1.0 mm厚度的鼻窦和胸膜处切片。37℃下，在含有2.5离子（KRB）（33）（另有说明）的橡胶盖烧瓶中进行孵育，或者对氨基)均匀标记的l -亮氨酸的研究j4c是放射性底物。α -氨基异丁酸-1 （IB）被用作非代谢底物来估计肝脏组织中氨基酸的积累。在14co2生成的研究中，d -葡萄糖-l- j4c、d -葡萄糖- 6j4c和棕榈酸-1- r4c与白蛋白络合（25）是底物。（所有放射性化学品都是从马萨诸塞州波士顿的新英格兰核能公司（New England Nuclear Corp.）购买的。）二氧化碳被困在0.4毫升的Hyamine氢氧化物中，放置在聚乙烯储存器中。在37℃孵育结束时，用针和注射器通过橡胶帽将二氧化碳吸收器放入储液器中。随后，向反应混合物中加入2.5 ml冷的20三氯乙酸（TCA），并将烧瓶置于室温下放置1小时。然后将中心孔置于含有10 ml甲苯的计数瓶中，其中含有1 mg/ml的PPO（2,5 -苯基恶唑）和0.25 mg/ml的POPOP (p-bis[2-(5-苯基恶唑-苯])，并进行Beckman LS-100液体闪烁系统计数。初步实验表明，该技术允许从Na14到克雷布斯-林格-比卡波的放射性
MATERIALS AND METHODSIncubation of lung slices. New Zealand albino male rabbits weighing about 2 kg were given food and water ad libitum3 sacrificed by stunning, and the lungs, heart, and adherent structures removed from the thorax en bloc. The external surface of the lungs were rinsed in cold 0.85 sliced at 0.5-1.0 mm thickness nchial structures and pleura. Incubations were erformed at 37 C in rubber-capped flasks containing 2.5 ion (KRB)(33) ss stated otherwise or studies on amino) uniformly labeled L-leucineJ4C was the radioactive substrate. Alpha-aminoisobutyric-1 IB) was used as a nonmetaboliz substrate to estimate mino acid accumulation by lu tissue.For studies on 14C02 production, D-glucose-l-J4C, D-glucose-6J4C, and palmitate-1-r4C acid complexed to albumin (25) were the substrates.(All radioactive chemicals were purchased from New England Nuclear Corp., Boston, Mass.) Carbon dioxide was trapped in 0.4 ml of hydroxide of Hyamine placed in a polyethylene reservoir. The CO2 absorber was placed in its reservoir, at the end of the 37 C incubation, through the rubber cap using needle and syringe. Following this, 2.5 ml of cold 20 trichloroacetic acid (TCA) were added to the reaction mixture and the flasks allowed to stand at room temperature for 1 hr. The center well was then placed in a counting vial with 10 ml of toluene, which contained 1 mg/ml of PPO (2, 5-diphenyloxazole) and 0.25 mg/ml of POPOP (p-bis [2-(5-phenyloxazolylj-benzene]), and was counted Beckman LS-100 liquid scintillation system. Prelirnishowed that this technique alw of the radioactivity from Na14 to the Krebs-Ringer-bicarbo