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目的:观察骨髓来源的细胞能否分化成肾小管上皮细胞。方法:15只绿色荧光蛋白(GFP)标记的C57BL/6转基因小鼠提供骨髓细胞,64只同种无荧光标记的C57BL/6小鼠随机分为正常对照组(N组)、全身照射组(TBI组)、骨髓移植组(BMT组)、骨髓移植+甘油注射组(B+G组),每组16只。各组小鼠于不同时间点取血检测血常规、尿素氮及血肌酐,并取肾行H-E染色检查肾脏病理变化。流式细胞仪可以明确受体鼠骨髓细胞中GFP阳性细胞的比例,利用荧光显微镜及激光共聚焦显微镜采用荧光组织化学、免疫组织化学等方法观察GFP阳性细胞在受体鼠肾脏的分布及数量。结果:致死剂量60Co照射虽然引起血常规三系减少,但并未对肾脏的组织结构及功能造成损伤。BMT组受体鼠在骨髓移植后第56天、84天时,肾小管中有少量GFP阳性细胞的存在,B+G组受体鼠于上述同样时间点时肾小管中的GFP阳性细胞增多。激光共聚焦显微镜进一步证实了这些GFP阳性细胞位于肾小管上皮,并且荧光组织化学显示,这些GFP阳性细胞表达肾小管上皮细胞特异性功能蛋白Megalin。结论:骨髓细胞可以向肾小管上皮细胞分化,参与肾小管上皮细胞的更新,并且损伤可以使骨髓细胞的肾向分化率增加。
Objective: To observe whether bone marrow-derived cells can differentiate into renal tubular epithelial cells. METHODS: Fifteen green fluorescent protein (GFP) -based C57BL / 6 transgenic mice were provided bone marrow cells. Sixty-four non-fluorescent C57BL / 6 mice were randomly divided into normal control group (N group), whole body irradiation group TBI group), bone marrow transplantation group (BMT group) and bone marrow transplantation + glycerol injection group (B + G group), 16 in each group. The mice in each group were sacrificed at different time points to detect the blood, urea nitrogen and serum creatinine, and renal pathological changes were examined by H-E staining. Flow cytometry can determine the percentage of GFP-positive cells in recipient bone marrow cells. Fluorescent and confocal microscopy were used to observe the distribution and quantity of GFP-positive cells in the kidney of recipient mice by fluorescence histochemistry and immunohistochemistry. Results: The lethal dose of 60Co irradiation caused by blood three lines decreased, but did not damage the structure and function of the kidney. In BMT group, there were a few GFP positive cells in renal tubules on the 56th day and 84th day after bone marrow transplantation. The GFP positive cells in tubular in the B + G group increased at the same time point. Laser confocal microscopy further confirmed that these GFP-positive cells were located in the renal tubular epithelium, and fluorescence histochemistry showed that these GFP-positive cells expressed tubular epithelial cell-specific functional protein Megalin. CONCLUSION: Bone marrow cells can differentiate into renal tubular epithelial cells and participate in the regeneration of renal tubular epithelial cells. Injury can increase the renal differentiation rate of bone marrow cells.