硫酸酯酶2型基因体内表达促进5-氟脲嘧啶诱发的小鼠骨髓抑制恢复作用研究

Protective Effect of Sulfatase 2 Gene Expression on Mice Myelodepression Induced by 5-Fluorouracil Injection

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。

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 estabhshed 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）×10^4/μl, n = 6 versus （ 14.7±2.1 ）×10^4/μl, n = 6, respectively, p 〈 0.05 ]. The total bone marrow ceils per single leg on day 7 in experimental group were （94.2±21.1 ）×10^4 （n = 6 ）, and significantly lower than the control group （ 173±59.9 ）×10^4 （ 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）×10^4（n =6） than in the control group （255±65.3 ）×10^4 （ 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.