电脉冲介导红细胞生成素基因肌肉转移治疗大鼠肾性贫血

研究电脉冲介导的质粒红细胞生成素基因肌肉转移效率以及对肾性贫血的治疗作用。方法用腺嘌呤诱导大鼠慢性肾性贫血模型,以电脉冲介导质粒ＥＰＯ基因肌肉转移对６０只大鼠实施基因治疗,治疗期间监测血红蛋白、红细胞压积血清肌酐、血清尿素氮,并用ＥＰＯ酶联免疫试剂盒测定血清ＥＰＯ水平,

The effects of electroporation-mediated erythropoietin (EPO) gene transfer into skeleton muscle on renal anemia

Objective　To investigate the effects of erythropoietin (EPO) gene transfer into skeleton muscle mediated by electroporation on renal anemia. Methods　Renal failure models were created by adenine-excessive diet (150 mg per day). Plasmid vectors encoding EPO were transferred by electroporation after 80 days when mean blood urea nitrogen level (BUN) had increased from 3.4 mmol/L±1.3 mmol/L to 18.1 mmol/L±4.1 mmol/L and the hematocrit had decreased from 45.6%±2.1% to 25.4%± 3.7%. During the process of treatment, adenine-excessive diet was given. Hb, HCT, BUN and Cre in blood were tested by automatic analyzer; EPO level in the serum was tested by EPO ELISA kit, EPO gene expression was proved by RT/PCR.The survival rate was caculated. Results　Hematocrit increased to 34.4%±7.5% only 7 days after the treatment and reached 91.4% of normal level (46%±2%) after 5 weeks. The survival rate of test models after 9 weeks was 77.8%, which was remarkably higher than that of controls (16.7%). mRNA level of EPO gene expression was indicated by RT/PCR. Conclusion　Electroporation can increase the efficiency of EPO gene transfer and thus greatly improve hematocrit in mice and prolong the life-span of chronic renal anemia models. This method can provide a new way for treatment of EPO-responsive anemias.