骨髓来源细胞移植对实验性肾小球肾炎小鼠的作用  

No Significant Benefits Can Bone Marrow Derived Cells Bring to Experiment Glomerular Nephritis of Mice

作　　者：张懿[1] 谌卫[1] 刘先锋[1] 房振宇[1] 于建平[1] 郭志勇[1] 梅长林[2] 程涛 

机构地区：[1]第二军医大学附属长海医院肾内科,上海200433 [2]第二军医大学附属长征医院肾内科,上海200003 [3]Hillman Cancer Center,University of Pittsburgh,Pittsburgh

出　　处：《中国中西医结合肾病杂志》2009年第5期382-385,I0001,共5页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：国家自然科学基金资助项目(No.30270620;30370664)

摘　　要：目的:利用骨髓移植的方法观察骨髓来源细胞在小鼠实验性肾小球肾炎中的作用。方法:以绿色荧光蛋白(GFP)标记的C57BL/6转基因小鼠为供体,同种无标记小鼠为受体鼠,受体鼠在接受供体骨髓细胞前经亚致死剂量^(60)Co照射,并于移植5周后以尾静脉注射的方式接受竹叶青蛇毒注射;蛇毒注射后7、14、28、56d利用免疫组织化学和免疫荧光双重染色方法观察受体鼠肾脏内的GFP阳性细胞及受体鼠肾脏病理变化,其中一组受体鼠在蛇毒注射后接受连续1周的粒巨系-集落刺激因子(GM-CSF)腹腔注射,观察GM-CSF对骨髓来源细胞及受体鼠的影响。结果:无论有无蛇毒损伤,荧光显微镜下受体鼠肾脏内均未见明亮的绿色荧光。GFP免疫组织化学可见所有受体鼠的肾小管上皮内均有少量细胞质棕褐色、细胞核大且紫蓝的GFP阳性细胞,包括那些骨髓移植后无蛇毒损伤的受体鼠;肾小球内也可见上述细胞,但在无蛇毒损伤的受体鼠内肾小球内未见上述细胞;激光共聚焦显微镜未见到明显双染色阳性的细胞,从形态上证实肾小球内的这些GFP阳性细胞大多为血细胞。那些接受GM-CSF注射的受体鼠肾脏内,GFP阳性细胞没有明显增加,而且肾脏损伤恶化,表现为蛇毒注射后1周出现了明显的局灶性节段性肾小球硬化。结论:骨髓来源细胞在蛇毒诱导的系膜增生型肾小球肾炎小鼠肾脏内向肾小管和肾小球细胞均可少量转化,但由于转化率低及肾小球结构的复杂性,向肾小球细胞的转化更加不易被识别;GM-CSF注射未能增加外源性骨髓细胞向肾脏实质细胞的转化而使受体鼠肾脏损伤恶化,于病程早期出现了局灶性节段性肾小球硬化。Objective： To investigate the role of bone marrow derived cells in the experiment glomerularnephritis of mice through the method of bone marrow transplantation. Methods：Green fluorescent protein （GFP） labeled C57BL/6 mice were donators, and homogeneity mice without GFP labeled were recipients, who accepted donators＇ bone marrow cells transplantation after 60Co sub - lethal dose irradiation, and five weeks after transplantation, received Habu - snake venom injection through vena caudalis. 7,14,28 and 56 days after the venom injection, GFP positive cells and pathological changes of the recipients＇ kidneys were investigated by immunohistochemistry and double staining immunofluorescence. Among the total recipients,some recipients in one group received GM - CSF intraperitoneal injection for successive one week after the venom injection to observe the effect of GM - CSF to the bone marrow cells and the recipients. Results： No bright green fluorescence was seen in all the recipients＇ kidneys no matter whether they were received the venom injection by fluorescence microscope or not. Immunohistochemistry of GFP showed that some foreign cells whose cytoplasm was brown and the nucleolus was dark blue and bigger than normal presented in tubule epithdium in all the recipients＇ kidneys even in those of no venom injection mice, some above-mentioned cells also presented in the recipients＇ glomerulus except those no venom injection mice, but these GFP positive cells were almost blood cells confirmed by morphology under confocal microscopy because they were not double staining positive cells. The number of GFP positive cells was not increased in those recipients who received GM- CSF injection, and the kidney injuries of these recipients were deteriorated as presented significant focal segmental glomerulosclerosis one Week after venom injection. Conclusion： Bone marrow derived cells can differentiate to tubules and glomerulus cells in a small quantity in snake venom induced mesenterium proliferative glomerulon

关 键 词：骨髓细胞 绿色荧光蛋白 竹叶青蛇毒 系膜增生型肾小球肾炎 

分 类 号：R692.3[医药卫生—泌尿科学]