Yun Jing-Ping, Liew Choong-Tsek, Chew Eng Ching, Yin Xiao-Yu, Lai Paul Bo San, Fai Yam Hin, Li H K Richard, Jin Mei-Lin, Ding Ming-Xiao, Li Ming-Tao, Lin Han-Liang, Lau Wan Yee
Department of Pathology, Cancer Center of Sun Yat-Sen University, Guangzhou, China.
J Cell Biochem. 2004 Apr 15;91(6):1269-79. doi: 10.1002/jcb.20034.
We explored the feasibility of studying nuclear matrix protein (NMP) expressions of the hepatocytes in normal and cirrhotic rat livers with liver regeneration after partial hepatectomy. Sixteen Wistar healthy rats were studied with experimental liver regeneration and/or liver cirrhosis. Two-dimensional (2-D) gel electrophoresis was used to generate these NMP compositions from these rat liver samples. Several antibodies against cytokeratin, vimentin, actin, B23, HNF4alpha, and heat shock protein 70 were used for identification by Western blot. Totally, 41 strongly stained protein spots were characterized on the 2-D gels. Thirty-four protein spots were detected in all of these rat livers, of which, cytokeratin, vimentin, actin, HNF4alpha, and heat shock protein 70 were identified. B23 was detected in the regenerated livers. Three protein spots (s33, s34, and s35) were detectable only in NMP preparation extracted from the regenerating rat livers after hepatectomy. Another three protein spots (s36, s37, and s38) were detectable only in NMP preparation extracted from thioacetamide-induced cirrhotic rat livers. Under these conditions including experimental liver regeneration and/or liver cirrhosis, Over thirty higher abundance NMPs of hepatocytes were consistently expressed and considered as common and basic NMPs. Some of the NMPs are specific for liver regeneration and may play a critical role in cell proliferation and cell cycle, and some are specific for liver cirrhosis.
我们探讨了研究部分肝切除术后正常和肝硬化大鼠肝脏中肝细胞的核基质蛋白(NMP)表达以及肝脏再生情况的可行性。对16只健康的Wistar大鼠进行了实验性肝脏再生和/或肝硬化研究。采用二维(2-D)凝胶电泳从这些大鼠肝脏样本中生成这些NMP成分。使用几种针对细胞角蛋白、波形蛋白、肌动蛋白、B23、肝细胞核因子4α(HNF4α)和热休克蛋白70的抗体,通过蛋白质免疫印迹法进行鉴定。在二维凝胶上总共鉴定出41个强染色的蛋白质斑点。在所有这些大鼠肝脏中检测到34个蛋白质斑点,其中鉴定出了细胞角蛋白、波形蛋白、肌动蛋白、HNF4α和热休克蛋白70。在再生肝脏中检测到了B23。仅在肝切除术后再生大鼠肝脏提取的NMP制剂中可检测到三个蛋白质斑点(s33、s34和s35)。另外三个蛋白质斑点(s36、s37和s38)仅在硫代乙酰胺诱导的肝硬化大鼠肝脏提取的NMP制剂中可检测到。在包括实验性肝脏再生和/或肝硬化的这些条件下,超过三十种丰度较高的肝细胞NMP持续表达,并被视为常见和基本的NMP。其中一些NMP对肝脏再生具有特异性,可能在细胞增殖和细胞周期中起关键作用,而一些则对肝硬化具有特异性。