干细胞因子联合粒细胞集落刺激因子对单侧输尿管梗阻大鼠骨髓干细胞及内皮祖细胞的动员作用

Mobilization effects of SCF along with G-CSF on bone marrow stem cells and endothelial progenitor cells in rats with unilateral ureteral obstruction

目的探讨干细胞因子(SCF)联合粒细胞集落刺激因子(G-CSF)对单侧输尿管梗阻(UUO)大鼠骨髓干细胞及内皮祖细胞的动员作用。方法56只Wistar大鼠随机分为7组:正常对照组、SCF组、G-CSF组、SCF联合G-CSF组(SCF-G组)、假手术组、UUO组、SCF联合G-CSF用于UUO组(UUO+SCF-G组)。实验第5天采集血标本后:①流式细胞仪检测静脉血单个核细胞中CD34+、CD34+/CD133+细胞;②检测血清谷丙转氨酶、谷草转氨酶、尿素氮、肌酐水平。结果①对照组大鼠静脉血单个核细胞中CD34+细胞为(0.13±0.01)%,假手术组CD34+细胞为(0.24±0.06)%与对照组比较差异无显著性(P>0.05);其余各组与对照组比较CD34+细胞百分率均明显增高(P<0.05),以UUO+SCF-G组(3.04±0.42)%及SCF-G组(2.10±0.28)%增高最为明显;②对照组大鼠静脉血单个核细胞中CD34+/CD133+细胞为(0.02±0.01)%,假手术组CD34+/CD133+细胞(0.05±0.02)%与对照组比较差异无显著性(P>0.05);其余各组与对照组比较CD34+/CD133+细胞百分率均明显增高(P<0.05),以UUO+SCF-G组(0.73±0.17)%增高最为明显;③7组血清尿素氮,肌酐、谷丙转氨酶水平无明显增高,UUO组谷草转氨酶水平较其他6组增高,差异有显著性。结论SCF和G-CSF对干细胞和内皮祖细胞的动员效果并非完全呈平行关系,联合使用可提高内皮祖细胞和干细胞的动员率,短期内未见肝、肾毒副作用。

Objective To study the mobilization effects of stem cell factor （SCF） along with granulocyte colonystimulating factor （G-CSF）on bone marrow stem cells and endothelial progenitor cells in rats with unilateral ureteral obstruction （UUO）. Methods Fifty-six healthy male Wistar rats were randomly divided into seven groups ： control, SCF, G-CSF, SCF ＋ G-SCF, Sham-operated, UUO and UUO ＋ SCF ＋ G-CSF groups （ n = 8 each）. The rats from the control, SCF, G-CSF and SCF ＋ G-CSF groups were hypodermically injected with normal saline （2 mL/kg）, SCF （200 μg/kg）, G-CSF （200 μg/kg） and SCF along with G-CSF respectively for 5 days. The rats from the UUO and UUO ＋ SCF ＋ G-CSF groups were subjected to the ligation of right ureter and then were hypodermically injected with normal saline （2 mL/kg） and SCF （200 μg/kg） ＋ G-CSF （200 μg/kg） respectively for 5 days. The sham-operated group had the same operative approach as the UUO and the UUO ＋ SCF ＋ G-CSF groups but the right ureter was not ligated. After operation they received a hypodermical injection of 2 mL/kg normal saline for 5 days. Five days later blood samples were collected. The percentages of CD34^＋ and CD34 ^＋/CD133 ^＋ cells in intravenous blood mononuclear cells were detected by flow cytometry. Serum contents of glutamate-pyruvate transaminase （ GPT）, glutamic oxalacetic transaminase （ GOT）, urea nitrogen and creatinin were measured. Results Except for the sham-operated group, the other five groups （ SCF, G-CSF, SCF ＋ G- SCF, UUO and UUO ＋ SCF ＋ G-CSF groups） had significantly higher percentage of CD34^＋ cells and CD34^＋/CD133 ^＋ cells in intravenous blood mononuclear cells than the control group （ P 〈 0. 05 ）. There were significant differences in the percentage of CD34 ^＋ cells and CD34 ^＋/CD133 ^＋ cells among the five groups （P 〈 0.05）. The UUO ＋ SCF ＋ G-CSF group showed the highest percentage of CD34 ^＋ cells and CD34 ^＋/CD133 ^＋ cells, followed by the SCF ＋ G-CSF group. There wereno significant differences in serum contents of GPT, urea nitrogen and creatinin among the seven groups. Except the UUO group showed higher GOT contents, there were no significant differences in the GOT contents among the other six groups. Conclusions The mobilization effects of SCF and G-CSF on bone marrow stem cells and endothelial progenitor cells were not always in paraller. A combination of SCF and G-CSF can effectively mobilize stem cells and endothelial progenitor cells, and side effects were not found in the liver and the kidney.