大鼠体内气管损伤修复过程及气管干细胞的定位研究  

THE WOUND-REPAIR PROCESS OF RAT TRACHEA IN VIVO AND THE LOCALIZATION OF TRACHEAL STEM CELLS

作　　者：廖爱军[1] 贾心善[2] 曲杨[2] 

机构地区：[1]中国医科大学附属第二医院血液内科,沈阳110004 [2]中国医科大学病理学教研室,沈阳110001

出　　处：《中国组织化学与细胞化学杂志》2005年第3期349-352,共4页Chinese Journal of Histochemistry and Cytochemistry

基　　金：国家自然科学基金资助项目(30170407)

摘　　要：目的观察大鼠体内气管损伤修复过程,进行气管干细胞的定位。方法应用氟尿嘧啶(5FU)诱发在体气管上皮损伤,动态观察修复过程;对损伤后气管上皮细胞行Hoechst33342荧光染色,并用RTPCR法检测ABC转运蛋白ABCG2/bcrp1基因。结果1.5FU作用30min后大鼠气管上皮细胞绝大部分脱落,可见少量间隔分布的类似裸核的细胞呈钉状位于基底膜上,免疫组化检测增殖细胞核抗原阴性,证明为G0期细胞。其中部分细胞Hoechst33342染色阴性,为侧群(sidepopulation,SP)细胞。2.将5FU去除3-6h后,上皮细胞形态变为扁平,9-12h后细胞变为立方,细胞数目逐渐增多,24h上皮细胞数更多,连接成片,可见纤毛,48h接近恢复假复层纤毛柱状上皮。3.RTPCR检测ABCG2/Bcrp1阳性反应产物长度为272bp。结论5FU打击后,残余的G0期气管上皮细胞中含有干细胞。Objective To explore the wound-repair process in vivo of rat tracheal epithelium induced by Fluorouracil (5-FU) and to localize tracheal stem cells. Methods The trachea was injured by 5-FU in vivo and the wound-repair process was dynamically observed under LM. Proliferating cell nuclear antigen (PCNA) expression was analyzed by immunohistochemistry (SP methods) and Hoechst33342 staining in injured tracheal epithelial cells was observed under fluorescence microscope. ABCG2/Bcrp1 was detected by RT-PCR. Results 1. Most epithelial cells exfoliated after a 30 minutes' treatment with 5-FU. A few nail-like cells which looked like naked nuclei were randomly distributed on the basement membrane. PCNA negative staining suggested that these were G_ 0 phase cells. Among them, there were some Hoechst33342 negative cells-SP (side population) cells. 2. After 5-FU was removed, the tracheal epithelium recovered. At 3-6h the cells became flat in morphology. At 9-12h, cuboidal and columnar epithelium appeared, and the number of the cells increased.At 24h, the epithelial cells connectedwith each other and the ciliatedcells conld be seen. Finally, at 48h the pseudostratified mucociliary epithelium was restored. 3.The product of ABCG2/Bcrp1(272bp)was detected by RT-PCR. Conclusion There were tracheal stem cells (SP cells) among G_ 0 phase cells which were left on the basement membrane after the injury by 5-FU.

关 键 词：气管 干细胞 氟尿嘧啶 ABC转运蛋白 

分 类 号：R364[医药卫生—病理学]