Pipkin J L, Hinson W G, Feuers R J, Lyn-Cook L E, Burns E R, Duffy P H, Hart R, Casciano D A
Department of Health and Human Services, Food and Drug Administration, Jefferson, AR 72079-9502.
Aging (Milano). 1994 Apr;6(2):121-31. doi: 10.1007/BF03324225.
A single intraperitoneal injection of the human therapeutic drug bleomycin (BL) was administered to three groups of male Fischer 344 rats at time 0, and the incorporation of [35S]methionine ("synthesis") and phosphorylation patterns of stress proteins (sps/hsps) from bone marrow cells were analyzed over time by two-dimensional electrophoresis and fluorography. Two groups of rats, young ad libitum (Y/AL--3 months) and old ad libitum (O/AL--28 months), had free access to rat chow, and a third group of old rats (O/CR--28 months) were maintained on a caloric restricted intake (60% of the AL diet). The administration of BL in Y/AL, O/AL and O/CR animals activated the 35S-labeling of sp 90 which reached a peak at 4 hours. Labeling of sp 90 was significantly greater in Y/AL compared to O/AL, and the incorporation pattern of O/CR was intermediate to Y/AL and O/AL animals. All labeling of sp 90 in each group had disappeared by 10 hours after BL administration. Stress protein 70x (inducible form) in these three animal groups displayed a similar pattern of 35S-incorporation, but the amount of labeling was less than that of sp 90. No labeling of sp 70x remained by 13 hours after BL administration. Phosphorylation ([32P] phosphate incorporation) of sp 90 reached a maximum level at 2 hours in all animals, and 32P-labeling in Y/AL was significantly increased over O/AL and O/CR with an intermediate level found in O/CR animals. The turnover rate (phosphorylation/dephosphorylation) of sp 90 induced by BL was significantly suppressed and temporarily extended in O/AL as compared with O/CR, which implied that CR not only increased incorporation of sp 90, but also enhanced a utilization of the phosphate pool very similar to that seen in Y/AL animals.
在时间点0对三组雄性Fischer 344大鼠进行单次腹腔注射人类治疗药物博来霉素（BL），并通过二维电泳和荧光自显影技术，随时间分析骨髓细胞中[35S]甲硫氨酸的掺入情况（“合成”）以及应激蛋白（sps/hsps）的磷酸化模式。两组大鼠，即自由摄食的年轻大鼠（Y/AL - 3个月）和自由摄食的老年大鼠（O/AL - 28个月），可自由获取大鼠饲料，第三组老年大鼠（O/CR - 28个月）维持热量限制摄入（自由摄食组饮食的60%）。在Y/AL、O/AL和O/CR动物中注射BL激活了sp 90的35S标记，该标记在4小时达到峰值。与O/AL相比，Y/AL中sp 90的标记显著更高，且O/CR的掺入模式介于Y/AL和O/AL动物之间。在注射BL后10小时，每组中sp 90的所有标记均消失。这三组动物中的应激蛋白70x（诱导型）呈现出相似的35S掺入模式，但标记量少于sp 90。在注射BL后13小时，sp 70x不再有标记。sp 90的磷酸化（[32P]磷酸盐掺入）在所有动物中于2小时达到最高水平，Y/AL中的32P标记相对于O/AL和O/CR显著增加，O/CR动物中处于中间水平。与O/CR相比，BL诱导的sp 90周转率（磷酸化/去磷酸化）在O/AL中显著受到抑制且暂时延长，这意味着热量限制不仅增加了sp 90的掺入，还增强了磷酸盐池的利用，这与Y/AL动物中所见非常相似。
