Evaluation of smooth muscle bioassays for prostaglandins E 1 and F 1 alpha.
Evaluation of smooth muscle bioassays for prostaglandins E 1 and F 1 alpha.
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前列腺素 E 1 和 F 1 α 平滑肌生物测定的评估。
DOI：
10.1152/jappl.1968.25.6.783
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发表时间：
1968
影响因子：
3.3
通讯作者：
W. E. Brown
中科院分区：
文献类型：
--
作者：
J. Weeks;J. R. Schultz;W. E. Brown
方法将所有肌肉固定在25-ml平滑肌浴（ME-5500，Metro Scientific Co.，Farmingdale，纽约）（3）.使用Grass FT在Grass测谎仪上记录收缩。03力传感器和等张弹簧”（兔十二指肠为1 g/cm，所有其他肌肉为0.3 g/cm）。在收缩范围内，与等渗性的偏差小于10%。传感器安装在支架上，支架可以通过转动螺杆垂直升降。测谎仪基线设置在50 mm宽图表底部上方5 mm处，将等于肌肉静息张力的砝码悬挂在传感器上，并将笔调整到该基线。在加入每种供试化合物之前，通过升高或降低传感器使注射笔达到该基线（即，使肌肉恢复至其初始静息张力）。这个过程模仿了传统的记波记录与肌肉杠杆。沙鼠结肠和大鼠胃底通常返回到初始基线，但豚鼠回肠，特别是兔十二指肠经常需要调整换能器，以使笔达到基线。然而，这些调整相对于收缩的幅度来说是很小的。一次最多测试四块肌肉。为了方便和均匀，暴露于测试化合物、洗涤和回收的时间由一个ERCE定时程序定时器（RC-5型，Industrial Timer Corporation，Parsipanny，纽约）控制，该定时器打开和关闭一个电磁阀（S90 A180 M型，Hoke，Inc.，克雷斯基尔，新泽西）在肌肉浴和盐水溶液储液器之间。在循环的洗涤部分期间，使100至125 ml的盐水溶液通过浴。选择暴露和恢复时间，以使化合物的作用最大化，并在清洗后松弛。在肌肉固定并稳定后，加入中等浓度的前列腺素并冲洗。包括该步骤是因为PGEr在几种平滑肌制剂中增强了多种激动剂，包括乙酰胆碱（参考文献和讨论见参考文献4）。然后，加入接近最大浓度的乙酰胆碱。当收缩最大时，调整多导生理记录仪的增益，使记录笔偏移高于基线40 mm。因此，所有随后的收缩都与乙酰胆碱的初始反应有关。对于兔十二指肠中的PGEl测试，测谎仪上的增益加倍，
METHODSGeneral. All muscles were mounted in 25-ml smooth muscle baths (ME-5500, Metro Scientific Co., Farmingdale, New York)(3). Contractions were recorded on a Grass polygraph with Grass FT. 03 force transducers and cCisotonic springs”(1 g/cm for rabbit duodenum, 0.3 g/cm for all other muscles). Over the range of contraction, the deviation from isotonicity was less than 10%. The transducers were mounted on supports which could be raised or lowered vertically by cranking threaded rods. The polygraph base line was set 5 mm above the bottom of the 50-mm wide chart, a weight equal to the resting tension for the muscle was hung on the transducer, and the pen adjusted to this base line. Before each test compound was added, the pen was brought to this base line by raising or lowering the transducer(ie, the muscle was returned to its initial resting tension). This procedure mimicked conventional kymographic recording with muscle levers. Gerbil colon and rat stomach fundus generally returned to the initial base line, but guinea pig ileum and especially rabbit duodenum frequently required adjustment of the transducer to bring the pen to base line. These adjustments, however, were small in relation to the magnitude of the contraction. Up to four muscles were tested at one time. For convenience and uniformity the time of exposure to test compounds, wash, and recovery was controlled by a singlecycle program timer (model RC-5, Industrial Timer Corporation, Parsipanny, New York) which opened and closed a solenoid valve (type S90A180M, Hoke, Inc., Cresskill, New Jersey) between the muscle bath and saline solution reservoir. From 100 to 125 ml of saline solution passed through the baths during the wash portion of the cycle. Exposure and recovery times were selected to allow maximal effect of the compound and relaxation after washing.After the muscles were mounted and allowed to stabilize, a midrange concentration of prostaglandin was added and washed out. This step was included since PGEr potentiates a variety of agonists, including acetylcholine, in several smooth muscle preparations(see ref. 4 for references and discussion). Then, a near-maximal concentration of acetylcholine was added. When the contraction was greatest the gain on the polygraph was adjusted so that the pen excursion was 40 mm above base line. Thus, all subsequent contractions were related to this initial response to acetylcholine. For tests on PGEl in rabbit duodenum, the gain on the polygraph was doubled so