CHARACTERIZATION OF THE EXTRACELLULAR BACTERICIDAL FACTORS OF RAT ALVEOLAR LINING MATERIAL
CHARACTERIZATION OF THE EXTRACELLULAR BACTERICIDAL FACTORS OF RAT ALVEOLAR LINING MATERIAL
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DOI：
10.1172/jci111537
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发表时间：
1984-01-01
影响因子：
15.9
通讯作者：
HSU, LC
中科院分区：
文献类型：
--
作者：
COONROD, JD;LESTER, RL;HSU, LC
无白细胞大鼠支气管肺泡灌洗液的表面活性剂部分（55,000 g 颗粒）含有快速杀死和溶解肺炎球菌的因子。通过使用定量杀菌测试来监测分级过程，对这些因子进行纯化和生化鉴定。用氯仿-甲醇（Bligh-Dyer 程序）提取大鼠表面活性剂后，在氯仿中恢复了大鼠表面活性剂 91% 的抗肺炎球菌活性。用氯仿、丙酮和甲醇进行硅酸层析后，所有可检测到的抗菌活性（约初始活性的80%）均用氯仿中的中性脂质洗脱。当用己烷、己烷-氯仿和氯仿在硅酸上重新色谱分析时，抗菌活性用游离脂肪酸(FFA)洗脱。 TLC 确定抗菌活性仅限于 FFA 部分。 GLC显示脂肪酸部分含有长链FFA(C12-C22)的混合物，其中66.7％是饱和的，32.4％是不饱和的。在标准化条件下（1 个杀菌单位）杀死 108 种肺炎球菌的 50% 所需的 TLC 纯化 FFA 的量为 10.6 +/-。 0.5 微克纯化的 FFA 充当去污剂，导致肺炎球菌细胞壁释放 [3H] 胆碱，并增加细菌细胞膜的通透性，3-O-[3H]甲基-D-葡萄糖的快速卸载证明了这一点。 FFA 作为去污剂似乎可以解释大鼠肺表面活性剂对肺炎球菌的杀菌和溶菌活性。
The surfactant fraction (55,000-g pellet) of leukocyte-free rat bronchoalveolar lavage fluid contains factors that rapidly kill and lyse pneumococci. These factors were purified and identified biochemically by using a quantitative bactericidal test to monitor fractionation procedures. Ninty-one percent of the antipneumococcal activity of rat surfactant was recovered in chloroform after extraction of rat surfactant with chloroform-methanol (Bligh-Dyer procedure). After chromatography on silicic acid with chloroform, acetone and methanol, all detectable antibacterial activity (.apprx. 80% of the initial activity) eluted with the neutral lipids in chloroform. When rechromatographed on silicic acid with hexane, hexane-chloroform and chloroform, the antibacterial activity eluted with free fatty acids (FFA). TLC, established that the antibacterial activity was confined to the FFA fraction. GLC showed that the fatty acid fraction contained a mixture of long-chain FFA (C12-C22) of which 66.7% were saturated and 32.4% were unsaturated. The quantity of TLC-purified FFA needed to kill 50% of 108 pneumococci under standardized conditions (1 bactericidal unit) was 10.6 .+-. 0.5 .mu.g. Purified FFA acted as detergents, causing release of [3H]choline from pneumococcal cell walls and increased bacterial cell membrane permeability, evidenced by rapid unloading of 3-O-[3H]methyl-D-glucose. FFA acting as detergents appear to account for the bactericidal and bacteriolytic activity of rat pulmonary surfactant for pneumococci.