动员自身骨髓干细胞与缺血再灌注肾损伤细胞的凋亡与增殖

Mobilization of autologous bone marrow stem cells is involved in cell apoptosis and proliferation following renal ischemia-reperfusion injuries

背景:骨髓干细胞具有多项分化潜能,可分化为肾组织固有细胞、修复损伤肾组织。目的:探讨粒细胞集落刺激因子联合干细胞因子动员自身骨髓干细胞对大鼠缺血再灌注肾损伤细胞凋亡与增殖的影响。方法:160只大鼠尿筛阴性后随机均分为正常对照组、模型组、细胞因子治疗组、治疗对照组。模型组和细胞因子治疗组建立大鼠单侧肾脏缺血再灌注损伤模型;细胞因子治疗组和治疗对照组于造模后24 h开始皮下注射粒细胞集落刺激因子(50μg/kg,1次/d)和干细胞因子(200μg/kg,1次/d),连续5 d;模型组不给药,正常对照组不予干预。TUNEL法检测细胞凋亡;免疫组织化学法(SABC法)检测肾组织CD34+细胞、Caspase-3、Bcl-2、细胞增殖核抗原表达情况。结果与结论:细胞因子治疗组肾组织内CD34+细胞较正常对照组、模型组明显增多(P<0.05)。不同时间点模型组和细胞因子治疗组凋亡指数、Capase-3表达量均高于正常对照组和治疗对照组(P<0.05),且模型组均显著高于细胞因子治疗组(P<0.05)。不同时间点模型组和细胞因子治疗组Bcl-2阳性表达细胞均高于正常对照组和治疗对照组(P<0.05)。细胞因子治疗组显著高于模型组,然后随着时间推移Bcl-2表达量明显减少(P<0.05)。模型组和细胞因子治疗组均可见细胞增殖核抗原阳性表达细胞;模型组于第24天增殖指数达峰值,后逐渐下降。细胞因子治疗组在第10天即达到高峰,持续至第17天,然后逐渐下降。说明粒细胞集落刺激因子联合干细胞因子动员自身骨髓干细胞可以促进肾缺血再灌注损伤后肾小管上皮细胞的增殖和减少细胞凋亡,从而有利于肾小管损伤的恢复。

BACKGROUND：Bone marrow stem cells are defined by their multi-potential ability, and can be differentiated into intrinsic cells in the kidney. OBJECTIVE：To study the effects of mobilizing autologous bone marrow stem cells by granulocyte colony-stimulating factor plus stem cellfactor on cellapoptosis and proliferation of rats with renal ischemia-reperfusion injury. METHODS：Total y 160 male Sprague-Dawley rats were randomly divided into four groups：control group, model group, cytokine treatment group, cytokine control group. Rat models of unilateral renal ischemia-reperfusion injury were established in the model and cytokine treatment groups. Rats in the cytokine treatment group and cytokine control group received subcutaneous injection of granulocyte colony-stimulating factor （50μg/kg） and stem cellfactor （200μg/kg）, once a day, for 5 continuous days. Rats in the model and control groups had no treatment. Apoptotic cells were detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling method, and the expression of CD34-positive cells, Caspase-3, Bcl-2, proliferating cellnuclear antigen in the kidney were measured using immunohistochemistry staining. RESULTS AND CONCLUSION：The number of CD34-positive cells in renal tissue of the cytokine treatment group was significantly higher than that of the control group and model group （P〈0.05）. The apoptotic index and expression of Capase-3 in the model group and cytokine treatment group were higher than those in the control group and cytokine control group （P〈0.05）. The apoptotic index and expression of Capase-3 in the cytokine treatment group were lower than that in the model group （P〈0.05）. The expression of Bcl-2 in the model group and cytokine treatment group was higher than that in the control group and cytokine control group （P〈0.05）. The expression of Bcl-2 in the cytokine treatment group was higher than that in the model group （P〈0.05）;however, as time went on, Bcl-2 expression was obviously decreased. Proliferating cellnuclear antigen expressed both in the model group and in the cytokine treatment group. Additional y, the proliferative index reached peak at 24 days in the model group, and then decreased gradual y;while in the cytokine treatment group, it reached the peak at 10 days, maintained a high level until the 17th day, and then decreased gradual y. Mobilization of autologous bone marrow stem cells by combination of granulocyte colony-stimulating factor and stem cellfactor can increase proliferation and decrease apoptosis of renal tubular epithelial cells after renal ischemia-reperfusion injury, and thus, promote the recovery from renal tubular injury.