MacIntyre S S, Schultz D, Kushner I
Biochem J. 1983 Mar 15;210(3):707-15. doi: 10.1042/bj2100707.
The synthesis and secretion of the acute-phase protein C-reactive protein by rabbit primary hepatocyte cultures was investigated. Hepatocytes prepared from animals that had received inflammatory stimuli 18-24 h before cell isolation were found to incorporate radiolabelled amino acids into C-reactive protein throughout the 48 h culture period. Intracellular C-reactive protein was found to be in steady state and there was no significant degradation of extracellular C-reactive protein, permitting direct estimation of rate of synthesis from rate of extracellular accumulation. Both C-reactive protein and total secreted protein were synthesized at constant rates for at least 24 h in culture. Mean rate of accumulation of newly synthesized total proteins in medium of cultures from six stimulated animals was 40% greater than was found in cultures from nine control (unstimulated) animals; this difference did not achieve statistical significance (0.05 less than P less than 0.10). Mean rate of C-reactive-protein synthesis represented 3.9% of total secreted-protein synthesis in cultures prepared from stimulated animals compared with 0.3% in cultures from control animals (P less than 0.001). Further, there was a correlation between C-reactive-protein synthesis by cultured hepatocytes and serum C-reactive-protein concentration at time of hepatocyte isolation (P less than 0.001). Rates of C-reactive-protein synthesis by hepatocyte cultures from stimulated animals were in good agreement with those previously measured in isolated perfused livers and those calculated from results of studies in vivo.
研究了兔原代肝细胞培养物中急性期蛋白C反应蛋白的合成与分泌。发现从细胞分离前18 - 24小时接受炎症刺激的动物制备的肝细胞,在整个48小时培养期内都将放射性标记的氨基酸掺入C反应蛋白中。发现细胞内C反应蛋白处于稳定状态,细胞外C反应蛋白没有明显降解,从而可以根据细胞外积累速率直接估算合成速率。在培养中,C反应蛋白和总分泌蛋白至少24小时以恒定速率合成。来自6只受刺激动物的培养物培养基中新合成的总蛋白的平均积累速率比来自9只对照(未受刺激)动物的培养物中高40%;这种差异未达到统计学显著性(0.05<P<0.10)。在受刺激动物制备的培养物中,C反应蛋白的平均合成速率占总分泌蛋白合成的3.9%,而在对照动物的培养物中为0.3%(P<0.001)。此外,培养的肝细胞合成C反应蛋白的速率与肝细胞分离时的血清C反应蛋白浓度之间存在相关性(P<0.001)。受刺激动物的肝细胞培养物中C反应蛋白的合成速率与先前在离体灌注肝脏中测得的速率以及根据体内研究结果计算出的速率高度一致。