X Lu1, A Zhou, C Jin. 1. Department of Biochemistry, Beijing Medical University, Beijing 100083, China.
Abstract
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 calculated. 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.
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 ureanitrogen 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 calculated. 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.