大鼠肝再生增强因子真核表达质粒的构建及其抗急性肝功能衰竭作用的研究

CONSTRUCTION OF EUKARYOTIC EXPRESS PLASMID OF RAT AUGMENTER OF LIVER REGENERATION GENE AND EFFECT ON ACUTE HEPATIC FAILURE IN RATS

目的研究肝再生增强因子真核表达质粒对急性肝功能衰竭的保护作用。方法PCR扩增的大鼠肝再生增强因子基因与真核表达载体 pcDNA3重组 ,重组后转化到大肠杆菌DH5α细胞内 ,扩增后提取重组的pcDNA3质粒。将重组的 pcDNA3质粒 (2 0 0 μg/kg)和生理盐水分别注射到 5 0 %CCL4复制的急性肝功能衰竭大鼠腹腔。在CCL4染毒后 4h开始注射 ,存活超过 96h计为存活 ,观察肝再生增强因子对大鼠急性肝功能衰竭的保护作用。结果 pcDNA3 ALR重组质粒目的基因与文献报道的大鼠ALRcDNA序列相同。pcDNA3 ALR重组质粒应用后 ,肝功能衰竭大鼠存活率 4 0 % ,明显高于生理盐水对照组的存活率 (1 5 % ,P <0 .0 1 )。

ObjectiveTo study the effect of eukaryotic express plasmid of augmenter of liver regeneration (ALR) on acute hepatic failure of rats. MethodsAfter the ALR gene was amplified using PCR, it was recombined with pcDNA3 plasmid. The recombinant plasmid DNA was used to treat the rats with acute hepatic failure. After the acute hepatic failure of rat was induced by peritoneal injection of 50% CCL 4 of 4 ml/kg, the rats were randomly divided into ALR therapy group and saline control group. The recombinant plasmid DNA (200 μg/kg) and saline were peritoneally injected into the rats with acute hepatic failure after 4 h of CCL 4 administration respectively. Rats living over 96 h were considered survival. ResultsThe sequence of ALR gene of recombinant plasmid of pcDNA3 ALR was accordant with the sequence of rat ALR cDNA reported. After the rats with acute hepatic failure were treated using the recombinant pcDNA3 ALR plasmid, the survival rate (40%) of rats was much higher than that of the rats in control group (15%, P< 0.01 ). ConclusionThe ALR gene can remarkably enhance the survival rate of rats with acute hepatic failure by promoting proliferation of hepatic cells.