骨髓细胞参与甘油引起的小鼠急性肾小管坏死修复的实验观察

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CFP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。

Effect of bone marrow cells in the recovery of acute tubular necrosis due to glycerine in mice

Objective:We aimed to investigate whether bone marrow cells can differentiate into renal tubular epithelial cells.Methodology:Fifteen transgenic C57BL/6 mice labeled with green fluorescent protein(GFP) served as bone marrow donors and sixty-four C57BL/6 mice without fluorescence labeled were randomly divided into 4 groups:normal group(N,n=16),total body irradiation group(TBI,n=16),bone marrow transplantation group(BMT,n=16),BMT plus glycerine intramuscular injection group(B G,n=16).The blood samples were obtained from these groups for routine analysis and examinations of urea nitrogen and plasma creatinine on the different times.Meanwhile,mice kidneys were harvested for H-E staining to observe the pathological changes.The amount of GFP positive bone marrow cells was determined by flow cytometry.The distribution and amount of GFP positive cells in renal tissues were observed by fluorescence histochemistry and immunohistochemistry used fluorescence microscope and laser scanning confocal microscope.Results:The lethally total body irradiation made no significant changes in renal histological structure and function though it made the red blood cells,white blood cells and platelet significant decrease.Some green fluorescence were seen in renal tubular epithelium of the recipients in BMI group on day 56 and 84 after BMT,while much green fluorescence were seen in renal tubular epithelium of the recipients in B G group on the same time.Furthermore,these cells were found in tubular epithelium by laser scanning confocal microscope,and fluorescence histochemistry,which indicated these cells express renal tubular epithelium specific protein-megalin.Conclusion:The bone marrow cells can differentiate into renal tubular epithelial cells and participate in regeneration of renal epithelium.The injury of renal tubular epithelial cells can enhance the rate of bone marrow cells transformation.