硫酸酯酶2型基因体内表达促进化疗药物诱发的小鼠骨髓抑制恢复作用研究

采用半定量RT-PCR和重组基因体内表达法观察了硫酸酯酶2基因（Sulfatase 2,Sulf2）在5-氟脲嘧啶（5-Fluorouracil, 5-Fu）诱发的小鼠骨髓抑制和再生过程中作用。结果表明：Sulf2在小鼠骨髓抑制和再生过程中呈现先上升，后下降的动态表达；电转pcDNA3.1-Sulf2基因实验组外周血白细胞数和血小板数在5-Fu注射后第7天分别为(1216.7±457.9)/μl和(8.1±5.4)万/μl，明显低于对照组［分别为(1691.7±228.9)/μl和(14.7±2.1)万/μl］，实验组单条腿骨髓细胞总数在第7天为(94.2±21.1)万，显著低于对照组（173±59.9）万，但在第11天为(585±337.9)万，又显著地高于对照组（255±65.3）万，实验组第7天10000个骨髓细胞总集落形成数为(9±8.4)，显著低于对照组（39±12.2)，统计均有显著性差异（p<0.05）。这些结果提示Sulf2可能对5 Fu诱发的小鼠骨髓抑制后的再生具有促进作用。

Promotive Effect of Sulfatase 2 Gene in vivo Expression during mouse bone marrow depression and regeneration induced by Chemotherapy

Abstract The aim of the study is to observe the regulation role of sulfatase 2 gene fragment in vivo expression on mouse myelodepression induced by 5-fluorouracil injection. The recombinant pcDNA3.1-Sulf2 eukaryotic expression vector was constructed using gene cloning method，mice bone marrow depression and regeneration model was established by injection of 5-fluorouracil (250mg/kg) via mice tail vein. 50μg of pcDNA3.1-Sulf2 recombinant eukaryotic expression vector (experimental group) was injected into the tibial muscle of each mouse, then introduced into muscle cells for expression by electroporation on the first day after 5-fluorouracil injection, so did the control group which injected and electroporated with the same quantity of pcDNA3.1 blank plasmid . Mice peripheral blood cells and platelets were calculated via automatic blood cells analyzer, and bone marrow cells per leg of each mouse was counted by hemacytometer on day 0 before injection of 5-fluorouracil and on day 3, day 7,day 11,day 14 after injection of 5-fluorouracil respectively. Colony forming test were carried out between the experimental group and control group on day 7 and day 14. The numbers of peripheral blood cells and platelets in experimental group on day 7 were significantly lower than those in the control group respectively ［(1216.7±457.9)μl, n=6 versus(1691.7±228.9)μl, n=6; and (8.1±5.4)×104/μl, n=6 versus (14.7±2.1)×104/μl, n=6, respectively, p<0.05］.The total bone marrow cells per single leg on day 7 in experimental group were (94.2±21.1)×104 (n=6)，and significantly lower than the control group (173±59.9)×104(n=6, p<0.05), but there was a significantly higher numbers of bone marrow cells per leg on day 11 in experimental group (585±337.9)×104(n=6) than in the control group (255±65.3)×104(n=6, p<0.05). Colony forming tests showed that the total colonies per 10000 of bone marrow cells on day 7 of experimental group was at 9±8.4 (n=4), significantly lower than that in control group 39±12.2(n=4, p<0.05), while there was no statistical significance between experimental group (23.3±7.6) per 1000 of bone marrow cells(n=4) and control group (28±6.1) per 1000 of bone marrow cells (n=3, p>0.05) on day 14. The results indicated that there is a protective effect of sulfatase 2 gene expression on the regeneration of mouse bone marrow depressed by 5-fluorouracil.