Miller L, Qureshi M A
Department of Poultry Science, North Carolina State University, Raleigh 27695-7608.
Poult Sci. 1992 Jun;71(6):988-98. doi: 10.3382/ps.0710988.
Synthesis of heat-shock proteins (HSP) in chicken macrophages, in response to thermal and nonthermal stressors, was determined. Cornell K-strain 6-wk-old White Leghorn females were injected with Sephadex and approximately 42 h later subjected to elevated temperatures in order to achieve a core body temperature (CBT) of 44 C. Peritoneal macrophages were isolated at 30 and 60 min after heat treatment. A parallel group of chickens, maintained at the normal CBT of 41 C, was used as controls and peritoneal macrophages were isolated after 60 min of treatment. For in vitro study of HSP response, cells of a chicken macrophage cell line (MQ-NCSU) were subjected to 45 C ambient temperature to produce heat shock (HS, thermal stress), lipopolysaccharide (LPS, 15 micrograms), and lead acetate (nonthermal stress) exposure for varying time periods. The HSP profiles of macrophages following various treatments were determined by one- and two-dimensional gel electrophoresis. The results showed that macrophages isolated from the 44 C CBT group synthesized HSP90, HSP70, HSP23, and a heat-inducible P32 protein. This HSP synthesis profile was similar to the HSP expression by MQ-NCSU cells exposed in vitro to 45 C conditions. Exposure to MQ-NCSU cells to lead acetate induced the same four proteins previously expressed by macrophages after in vivo or in vitro heat treatment. Two-dimensional analysis of lysates from cells treated with LPS, HS, or LPS plus HS treatments revealed a doublet protein molecule (70a and 70b) with identical molecular mass of 70 kDa. However, the pI value (isoelectric point) of 70b was higher (5.1) than that of 70a, which, along with HSP90 and HSP23, focused more toward the acidic side with a pI value of less than 4.6. The present study is the first to report pI profiles of chicken macrophage HSP. The in vitro and in vivo studies suggest that chicken macrophages respond to thermal and nonthermal stressors by producing similar kinds of "stress proteins".
研究了鸡巨噬细胞在热应激和非热应激条件下热休克蛋白（HSP）的合成情况。选用6周龄康奈尔K系白来航雌性母鸡，注射葡聚糖，约42小时后使其体温升高，以达到44℃的核心体温（CBT）。热处理后30分钟和60分钟分离腹腔巨噬细胞。将一组维持在正常核心体温41℃的鸡作为对照，处理60分钟后分离腹腔巨噬细胞。为了体外研究热休克蛋白反应，将鸡巨噬细胞系（MQ-NCSU）的细胞置于45℃环境温度下进行热休克（HS，热应激）、脂多糖（LPS，15微克）和醋酸铅（非热应激）处理，处理时间不同。通过一维和二维凝胶电泳确定各种处理后巨噬细胞的热休克蛋白谱。结果表明，从44℃核心体温组分离的巨噬细胞合成了HSP90、HSP70、HSP23和一种热诱导P32蛋白。这种热休克蛋白合成谱与体外暴露于45℃条件下的MQ-NCSU细胞的热休克蛋白表达相似。醋酸铅处理MQ-NCSU细胞诱导出与体内或体外热处理后巨噬细胞先前表达的相同的四种蛋白。对用LPS、HS或LPS加HS处理的细胞裂解物进行二维分析，发现一种双重蛋白分子（70a和70b），分子量相同，均为70 kDa。然而，70b的等电点（pI值）较高（5.1），高于70a，70b与HSP90和HSP23一起，更集中在酸性一侧，pI值小于4.6。本研究首次报道了鸡巨噬细胞热休克蛋白的pI谱。体外和体内研究表明，鸡巨噬细胞通过产生相似种类的“应激蛋白”对热应激和非热应激作出反应。
