Objective: To investigate the thrombopoietic effect of recombinant human thrombopoietin (rhTpo) gene transferred by electric pulse to normal and experimental thrombocytopenia mice.
Methods: Eukaryotic high expressing plasmid pcDI/Tpo was constructed by gene recombinant technology. 200 microg of the recombinant plasmid was injected into quadriceps femoris muscle of normal and experimental thrombocytopenia mice. Six times of electric pulse at 100v, 1Hz, 40ms were given immediately after the injection.The expression of rhTpo gene and its protein were assayed by RT-PCR and Western Blotting, respectively. Serum Tpo concentration was assayed by ELISA method.
Results: The recombinant plasmid pcDI/Tpo was successfully constructed. The expression of mRNA and protein of rhTpo gene was detected in the skeletal muscle of mice after transfection. Serum Tpo level increased from 328 +/- 89 ng/L to 1185 +/- 264 ng/L, and the platelet level of transfected mice increased from (259 +/- 27) x 10(9)/L to (640 +/- 31) x 10(9)/L. After injection with carboplatin, the platelet level decreased, but the nadir point was higher in pcDI/Tpo group than that in control group, and the recovery time of platelet count in pcDI/Tpo group shortened.
Conclusion: The rhTpo gene could be effectively transfected to mice by electric pulse and played thrombopoietic role in vivo.