| 摘要: |
| 目的：体外重组表达人肝再生增强因子（augmenter of liver regeneration，ALR)并进行功能研究。方法：全基因合成ALR序列并插入原核表达载体pET28a+ ，转化入BL21菌内进行诱导表达，表达产物经过纯化后利用MTT法观察表达产物对人肝细胞的刺激增殖活性；利用小鼠急性四氯化碳损伤模型观察表达产物的肝功能保护作用。结果：酶切鉴定及测序结果提示表达产物正确；纯化后表达产物具有明确的促进肝细胞增殖作用，并在中、低剂量具有降低小鼠急性化学性损伤后转氨酶水平的作用。结论：重组表达的ALR结构正确，具有促进肝细胞再生的作用。 |
| 关键词: 肝再生增强因子 DNA，重组 肝细胞 细胞增殖 转氨酶类 |
| DOI：10.3724/SP.J.1008.2007.01284 |
| 投稿时间：2007-09-21修订日期：2007-10-18 |
| 基金项目:国家自然科学基金（30571774）. |
|
| Genetic optimization, recombinant expression and functional study of human augmenter of liver regeneration |
| HUANG Ying-feng1，2,ZHOU Fei-guo3, GAO Chun-fang1*, WANG Hao4, GAO Zhi-yuan1, WANG Kun1, CHEN Jie2 |
| (1.Department of Laboratory Medicine, Eastern Hepatobiliary Hospital, Second Military Medical University, Shanghai 200438, China; 2.Shanghai You-Heng Biotech Inc., Shanghai 201413; 3.Department of Hepatic Surgery, Eastern Hepatobiliary Hospital, Second Military Medical University, Shanghai 200438; 4.Department of Laboratory Medicine, Changzheng Hospital, Second Military Medical University, Shanghai 200003) |
| Abstract: |
| Objective：To construct recombinant expression vector of human augmenter of liver regeneration (ALR) and study its protective effect on liver function. Methods: ALR cDNA was synthesized and inserted into expression vector pET28a+. The recombinant plasmid was tranformed into BL21 and the expression of ALR was induced by isopropyl-β-D-thiogalactoside(IPTG). MTT method was used for cell proliferation assay; the protective effect of recombinant product on liver function was observed in CCl4-induced acute toxic mouse model. Results: Recombinant expression plasmid of ALR was confirmed correct by restriction enzyme digestion and sequencing. The purified expression product had strong stimulatory effect on hepatocyte proliferation. Low and medium dosages of expression product decreased aminotransferase level in acute chemical injury mouse model.Conclusion: The recombinant expression vector of ALR has been correctly constructed and the expressed rALR can simulate hepatocyte regeneration. |
| Key words: augmenter of liver regeneration DNA,recombinant hepatocytes cell proliferation transaminases |
