Effect of temperature and inhibitors on serotonin-14C release from human platelets.
Effect of temperature and inhibitors on serotonin-14C release from human platelets.
复制标题
温度和抑制剂对人血小板 5-羟色胺-14C 释放的影响。
DOI：
10.1152/ajplegacy.1971.220.1.105
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发表时间：
1971
期刊：
影响因子：
--
通讯作者：
M. Zucker
中科院分区：
文献类型：
--
作者：
J. F. Valdorf;M. Zucker
材料和方法14 C-标记的血清素肌酸酐硫酸盐，约40 pc/pmole（Nuclear-Chicago Corp.，伊利诺伊州芝加哥），溶于8 ml的70%乙醇中，并保持在-20 ℃。除非另有说明，否则将聚集剂和抑制剂溶于0.15 M NaCl中。我们使用：腺苷5-二磷酸（ADP）（d 'rso rum salt）和腺苷（Sigma d' Chemical Co.，St. Louis，MO.）;盐酸肾上腺素（1 mg/ml，Parke，Davis & Company，Detroit，MI）;乙酰水杨酸（阿司匹林）（Amend Drug and Chemical Co.，纽约市）;乙二胺四乙酸二钠（Na 2-EDTA）（GeigyPharmaceuticals，Ardsley，NY）; N-乙基马来酰亚胺（MannResearchLaboratories，纽约市）;和二胍基二苯砜（由MerckSharp & Dohme，西点，PA，R博士巴勒斯·惠康公司的巴茨利，塔卡霍，纽约）。胶原蛋白，由Armour Pharmaceutical Company，坎卡基，IL.的William托马斯博士友情提供，是用乙醚、10%NaCl和缓冲液（28）提取的牛皮真皮残渣。用Teflon研杵手动均质化5 mg/ml的储备混悬液。高度纯化的人凝血酶由弗拉迈阿密的国家儿童心脏病医院的肯特米勒博士制备，并作为500 U/ml 0.1%人白蛋白的储备溶液储存在-20 ℃。用NaOH中和一碘乙酸制备碘乙酸钠。受试者为健康个体，至少1周未服用阿司匹林或含阿司匹林的药物。除非另有说明，否则通过16号针头和塑料管抽取的血液采集在九分之一体积的3.2%柠檬酸钠溶液中。其它抗凝剂是酸-柠檬酸盐-葡萄糖，配方A（> $ th体积）;柠檬酸盐-磷酸盐-葡萄糖（第36体积）;酸化柠檬酸盐（第36体积）（1）;或肝素（Lipo-Hepin，RikerLaboratories，Northridge，CA，100 U/ml等渗盐水，第36体积）。通过在室温下缓慢离心制备富血小板血浆（PRP），并置于融冰中。初步实验表明，在板没有变化-
MATERIALS AND METHODS14C-labeled serotonin creatinine sulfate, about 40 pc/pmole (Nuclear-Chicago Corp., Chicago, Ill.), was dissolved in 70% alcohol at 8 &ml and kept at-20 C. Unless otherwise noted, aggregating agents and inhibitors were dissolved in 0.15 M NaCl. We used: adenosine 5-diphosphate(ADP)(d’rso rum salt), and adenosine (Sigma d’Chemical Co., St. Louis, MO.); epinephrine HCl (1 mg/ml, Parke, Davis & Company, Detroit, Mich.); acetylsalicylic acid (aspirin)(Amend Drug and Chemical Co., New York City); disodium ethylenediaminetetraacetate(Na 2-EDTA)(Geigy Pharmaceuticals, Ardsley, NY); N-ethylmaleimide (Mann Research Laboratories, New York City); and diguanidinodiphenylsulfone(obtained through the courtesy of Merck Sharp & Dohme, West Point, Pa., and Dr. R. Baltzly of Burroughs Wellcome & Co., Tuckahoe, NY). Collagen, kindly provided by Dr. William Thomas of Armour Pharmaceutical Company, Kankakee, Ill., was the residue of bull-hide corium extracted with ether, 10% NaCl, and buffers (28). A stock suspension of 5 mg/ml was homogenized by hand with a Teflon pestle. Human thrombin, highly purified, was prepared by Dr. Kent Miller of the National Childrens Cardiac Hospital, Miami, Fla., and stored at-20 C as a stock solution of 500 U/ml 0.1% human albumin. Sodium iodoacetate was prepared by neutralizing monoiodoacetic acid with NaOH. The subjects were healthy individuals who had taken no aspirin or aspirin-containing drugs for at least 1 week. Blood, drawn through a 16-gauge needle and plastic tubing, was collected in one-ninth volume of 3.2% sodium citrate solution unless otherwise noted. Other anticoagulants were acid-citrate-dextrose, Formula A (> $ th volume); citratephosphate-dextrose(36th volume); acidified citrate (36th volume)(1); or heparin (Lipo-Hepin, Riker Laboratories, Northridge, Calif., 100 U/ml isotonic saline, 36th volume). Platelet-rich plasma (PRP) was prepared by slow centrifugation at room temperature and placed in melting ice. Preliminary experiments revealed no change in the plate-