外源性肝再生增强因子重组质粒在大鼠肝组织中的表达研究  

Study of the expression of exogenous augmenter of liver regeneration recombinant plasmid in liver tissue of rat

作　　者：李青[1] 刘殿武[1] 何海艳[1] 肖永红[1] 刘英丽[1] 

机构地区：[1]河北医科大学流行病学教研室,石家庄050017

出　　处：《卫生研究》2007年第1期66-68,共3页Journal of Hygiene Research

基　　金：河北省自然基金资助项目(No.302489)

摘　　要：目的研究外源性肝再生增强因子(ALR)重组质粒在免疫性肝纤维化大鼠肝组织中的表达情况。方法猪血清腹腔注射制备免疫性肝纤维化大鼠模型,pcDNA3-ALR重组质粒静脉注射对其进行干预,通过免疫组化、Western bolting检测大鼠肝组织ALR蛋白质的表达情况,通过RT-PCR检测ALR mRNA的表达情况。结果所有组的大鼠肝组织均有ALR蛋白的表达,其中,以正常组表达最弱,ALR处理组表达最强。图像分析细胞染色密度,正常对照组与模型组的结果分别为(0.109±0.01)和(0.159±0.02),二者相比差异有显著性(P<0.01);ALR处理组为(0.198±0.04),与模型组相比差异也有显著性(P<0.01)。Western bloting测定肝组织ALR的表达的结果与免疫组化相似。ALR mRNA在各组大鼠肝组织的表达pcDNA3-ALR处理组在约550bp处出现了用特定引物扩增的特异基因条带,此条带与直接从大肠杆菌中制备的pcDNA3-ALR重组质粒的扩增条带相同。正常对照组和模型组未出现用特定引物扩增的条带。结论ALR在正常大鼠肝组织的表达较低,在免疫损伤后的肝组织的表达明显增强。ALR重组质粒可在免疫性肝纤维化大鼠肝组织中发生表达,pcDNA3-ALR重组质粒处理组ALR蛋白表达明显增强是由于外源性ALR与内源性ALR表达“叠加”的结果。Objective To observe the expression of exogenous ALR recombinant plasmid in liver tissue of rats suffered from immune hepatic fibrosis. Methods Immune hepatic fibrosis model of rats were made by using porcine serum, which were treated by pcDNA3-ALR recombinant plasmid. The expression of ALR proteins in liver tissue were detected by immunohistochemistry staining and Western blotting and the expression of ALR mRNA were detected by RT-PCR. Results ALR were expressed in liver tissue in all group rats. ALR expressions were lower in normal control groups , and were higher in ALR treat groups. The results of image analysis by immunohistochemistry were （0. 109 ± 0.01 ）, （0. 159 ± 0.02） and （0.198 ± 0.04） in normal groups, model groups and ALR groups respectively. There were significant differences between model and normal groups （ P 〈 0.01 ） as well as ALR groups and model groups （ P 〈 0.01 ） . The results of Western blotting were similar to immunohistochemistry. The specific 550bp gene fragments that were amplified by RT-PCR with a couple of especial primers were found in ALR group but not in normal and model groups, which was coincident with fragments amplified by PCR with ALR recombinant plasmid extracted direcdy from competent E. coli DHS. Conclusion The expression of ALR was relatively low abundance in normal rat liver tissue, and increased markedly in immune injured rat liver tissue. The expression of the pcDNA3-ALR recombinant plasmid can be found in rat liver tissue suffered from immune hepatic fibrosis. The reinforcements of expression of ALR protein in liver tissue of pcDNA3-ALR recombinant plasmid treatment group was resulted from the overlap of endogenous and exogenous ALR expression.

关 键 词：肝纤维化 肝再生增强因子 基因表达 大鼠 

分 类 号：R575[医药卫生—消化系统] Q593.1[医药卫生—内科学]