| 摘要: |
| 目的:观察生理情况下骨髓细胞能否分化成肾小管上皮细胞.方法:绿色荧光蛋白(GFP)标记的C57BL/6转基因小鼠提供骨髓,同种无荧光标记的C57BL/6小鼠分为3组,每组各15只.正常对照组未做任何处理;全身照射组经致死剂量γ射线全身均匀照射后不接受骨髓移植,1周内处死;骨髓移植组照射后接受骨髓移植,分别于移植后56 d、84 d分2批处死小鼠.取3组小鼠血液行血常规、血肌酐、尿素氮检测,同时取其肾脏标本行H-E染色.流式细胞仪检测骨髓移植组骨髓细胞中GFP阳性细胞比例,免疫组织化学法观察GFP阳性细胞在其肾脏内的分布及数量.结果:与正常对照组相比,放射组和骨髓移植组的血尿素氮和肌酐无明显变化.肾脏病理检查示全身致死剂量γ射线照射未造成小鼠肾脏组织结构和生理功能的明显改变,但血常规提示其三系减少,与其他2组相比有统计学差异(P＜0.05).荧光显微镜和激光共聚焦显微镜发现骨髓移植后56、84 d的受体鼠肾小管中有少量GFP阳性细胞存在,流式细胞仪检测发现其骨髓中GFP阳性细胞的比例分别为(78.75±5.99)％和(79.58±4.60)％,免疫组化定量结果显示GFP阳性细胞所占肾小管上皮细胞的比例分别为(4.17±1.66)％和(3.88±1.35)％,两亚组间比较无明显差别.结论:在无肾小管损伤的生理情况下,骨髓细胞也可以参与肾小管上皮细胞的更新. |
| 关键词: 骨髓细胞、肾小管、绿色荧光蛋白、激光共聚焦显微镜 |
| DOI：10.3724/SP.J.1008.2006.00607 |
| 投稿时间：2006-03-01修订日期：2006-06-02 |
| 基金项目:国家自然科学基金（30270620,30370664）. |
|
| Role of bone marrow cells in renewal of renal tubular epithelial cells under physiological condition |
| 李保春,张懿,刘先峰,崔若兰,程涛,LI Bao-chun,ZHANG Yi,LIU Xian-feng,CUI Ruo-lan,CHENG Tao |
| () |
| Abstract: |
| Objective：To observe whether bone marrow cells （BMCs） can differentiate into the renal tubular epithelial cells under physiological condition. Methyls： Transgenic C57BL/6 mice labeled with green flouresent protein （GFP） served as bone marrow donors and C57BL/6 mice without fluorescence labeling were divided into 3 groups： normal group （N, n = 15）, total body irradiation group （TBI, n= 15）, and bone marrow transplantation （BMT, n= 15） group. The mice in Group N received no treatment, those in Group TBI were irradiated with lethal dose of γ rays but received no transplant （sacrificed within 1 week after irradiation）, and those in Group BMT were transplanted with BMCs after the irradiation （sacrificed at 58 and 84 days after transplantation）. Blood samples were obtained from all 3 groups for routine analysis and analysis of urea nitrogen and plasma creatinine. Meanwhile, mice kidneys were harvested for H-E staining. The amount of GFP positive BMCs in mice of BMT group was determined by flow cytometry; the distribution and amount of GFP positive cells in renal tissues were observed by immunohistochemistry. Results： There was no difference in the levels of urea nitrogen and plasma creatinine between the 3 groups. Lethal-dose total body irradiation resulted in no significant change in renal histological structure and function, but RBC, WBC and PLT of the mice in TBI group were markedly reduced compared with those in the other 2 groups （P〈0.05）. Green fluorescence was seen in renal tubular epithelium of mice on 56 and 84 days after transplantation by fluorescent microscope and laser scanning confocal microscope. Flow cytometry result showed that GFP positive BMCs were （78. 75 ± 5. 99）% and （79. 58±4.60）% in recipient mice on 56 and 84 days after transplantation, respectively; and immunohistochemistry staining showed that GFP positive cells in renal tubular epithelial of the recipient mice were （4.17±1.66）% and （3. 88±1.35）%, respectively （P〉0.05）. Conclusion： Bone marrow cells may participate in renewal of tubular epithelial cells even without the renal tubular damage. |
| Key words: bone marrow cells renal tubule green fluorescent protein laser scanning confocal microscope |
