Lee Ae Kyung, Kang Keon Wook, Kim Yoon Gyoon, Cho Min Kyung, Lee Myung Gull, Shim Chang-Koo, Chung Suk Jae, Kim Sang Geon
College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University, South Korea.
Mol Cell Biochem. 2002 Feb;231(1-2):163-71. doi: 10.1023/a:1014481319278.
Protein-calorie malnutrition (PCM), as one of global health problems, arises during protein and/or energy deficit due to disease and nutritional inadequacy. Previously, we showed that PCM elicited oxidative stress with activation of the phase II detoxifying gene expression, which was reversed by cysteine supplementation. As part of the attempts to identify the cellular adaptive responses and the associated gene expression during PCM, the current study was initiated to analyze the genes differentially expressed in the rat during PCM. Among 1,916 bands amplified, 85 putative differentially amplified bands were enhanced by PCM in the liver, while the expression of 64 bands was suppressed. Northern and/or reverse transcription-polymerase chain reaction (RT-PCR) analyses revealed that PCM increased the expression of fibrinogen B beta chain, B cell translocation gene I (BTGI) and thyroid hormone responsive protein (THRP) mRNAs. The increase in the hepatic fibrinogen B beta chain mRNA was not prevented by cysteine supplementation, whereas cysteine decreased the enhancement in the rGSTA2 and microsomal epoxide hydrolase mRNA expression. Cysteine was also active in reversing the increase in BTG1 mRNA during PCM. This was supported by the increase in BTG1 mRNA in H4IIE cells exposed to sulfur amino acid-deprived medium. Northern blot analysis revealed that THRP, highly expressed in the brain in a tissue-specific manner, was induced by PCM and that cysteine supplementation abolished the THRP induction. Conversely, the level of hepatic albumin mRNA was markedly decreased by PCM, which was partially restored by cysteine supplementation. Differential display RT-PCR analysis allowed us to identify the genes that are responsive to oxidative stress during PCM and to characterize the differential role of cysteine on the expression of the fibrinogen B beta chain, BTG1 and THRP genes as a homeostatic adaptive response during protein deficiency.
蛋白质 - 热量营养不良（PCM）作为全球健康问题之一，是由疾病和营养不足导致蛋白质和/或能量缺乏时出现的。此前，我们发现PCM通过激活II期解毒基因表达引发氧化应激，而补充半胱氨酸可逆转这种情况。作为识别PCM期间细胞适应性反应及相关基因表达的部分尝试，本研究旨在分析PCM大鼠中差异表达的基因。在扩增的1916条条带中，85条假定的差异扩增条带在肝脏中因PCM而增强，而64条条带的表达受到抑制。Northern印迹和/或逆转录 - 聚合酶链反应（RT-PCR）分析表明，PCM增加了纤维蛋白原Bβ链、B细胞易位基因I（BTG1）和甲状腺激素反应蛋白（THRP）mRNA的表达。补充半胱氨酸并不能阻止肝脏中纤维蛋白原Bβ链mRNA的增加，而半胱氨酸可降低rGSTA2和微粒体环氧化物水解酶mRNA表达的增强。半胱氨酸在逆转PCM期间BTG1 mRNA的增加方面也有作用。暴露于硫氨基酸缺乏培养基的H4IIE细胞中BTG1 mRNA的增加支持了这一点。Northern印迹分析显示，THRP在大脑中以组织特异性方式高度表达，由PCM诱导，补充半胱氨酸可消除THRP的诱导。相反，PCM使肝脏白蛋白mRNA水平显著降低，补充半胱氨酸可部分恢复。差异显示RT-PCR分析使我们能够识别PCM期间对氧化应激有反应的基因，并表征半胱氨酸对纤维蛋白原Bβ链、BTG1和THRP基因表达的不同作用，作为蛋白质缺乏期间的一种稳态适应性反应。
