Carpenter K L, Ball R Y, Carter N P, Woods S E, Hartley S L, Davies S, Enright J H, Mitchinson M J
Department of Pathology, University of Cambridge, U.K.
Adv Exp Med Biol. 1989;266:333-43. doi: 10.1007/978-1-4899-5339-1_24.
Mouse resident peritoneal macrophages (MPM) cultured with artificial lipoprotein consisting of cholesteryl linoleate complexed with bovine serum albumin (CL/BSA) rapidly accumulate ceroid in the form of rings. Experiments with various phenolic radical scavenger antioxidants and derivatives showed that the radical scavengers which are strongly lipophilic, and possess a free (i.e. non-esterified) phenolic hydroxyl group are inhibitors of ceroid ring formation. Time-course experiments with MPM and CL/BSA in which either or both of the components of the artificial lipoprotein have been oxidised before feeding showed that such oxidation accelerated ceroid accumulation, and suggested that oxidation of the lipoprotein is rate-determining in ceroid accumulation. Copper appeared to be a good catalyst for this. Agents able to activate the respiratory burst production of reactive oxygen species appeared to have no accelerating effect on ceroid accumulation from CL/BSA by MPM in a time-course. A novel method has been attempted for quantitating ceroid in MPM by means of its autofluorescence, using a Fluorescence-activated Cell Sorter (FACS). The results from FACS agree qualitatively with those from alcohol-xylene treatment followed by oil-red-o staining (AX/ORO). MPM cultured with CL/BSA for up to 4 days showed a 2.7-4.6-fold increase in mean fluorescence (at wavelengths greater than 490 nm) over MPM cultured with cholesteryl oleate/BSA (CO/BSA), with CL/BSA/butylated hydroxytoluene (CL/BSA/BHT), with CL/BSA/probucol, and with no artificial lipoprotein. The implications of the findings with respect to human atherosclerosis are discussed.
用由与牛血清白蛋白复合的亚油酸胆固醇（CL/BSA）组成的人工脂蛋白培养的小鼠腹膜常驻巨噬细胞（MPM）会迅速以环状形式积累类脂褐素。用各种酚类自由基清除剂抗氧化剂及其衍生物进行的实验表明，具有强亲脂性且带有游离（即未酯化）酚羟基的自由基清除剂是类脂褐素环形成的抑制剂。在用MPM和CL/BSA进行的时程实验中，在投喂前人工脂蛋白的一种或两种成分已被氧化，结果表明这种氧化加速了类脂褐素的积累，并表明脂蛋白的氧化在类脂褐素积累中起速率决定作用。铜似乎是对此的良好催化剂。能够激活活性氧呼吸爆发产生的试剂在时程上似乎对MPM从CL/BSA中积累类脂褐素没有加速作用。已经尝试了一种通过其自发荧光，使用荧光激活细胞分选仪（FACS）对MPM中的类脂褐素进行定量的新方法。FACS的结果在定性上与乙醇 - 二甲苯处理后油红O染色（AX/ORO）的结果一致。用CL/BSA培养长达4天的MPM与用油酸胆固醇/BSA（CO/BSA）、CL/BSA/丁基羟基甲苯（CL/BSA/BHT）、CL/BSA/普罗布考培养的MPM以及未用人工脂蛋白培养的MPM相比，平均荧光（在波长大于490nm处）增加了2.7 - 4.6倍。讨论了这些发现对人类动脉粥样硬化的意义。
