骨髓间充质干细胞和干细胞因子在老年炎症性肠病的实验研究  

Bone marrow mesenchymal stem cells and stem cell factor in the experimental study of inflammatory bowel disease

作　　者：辛国荣[1] 倪健[1] 刘洋[1] 吉志武[1] 赵斌[1] 于海涛[1] 朱晓文[1] 杨泽[2] 陈福军[1] 

机构地区：[1]佳木斯大学附属第一医院肛肠科,154002 [2]北京医院北京老年医学研究所,100730

出　　处：《中国老年保健医学》2017年第5期3-11,共9页Chinese Journal of Geriatric Care

基　　金：国家自然科学基金(81460203;8321120527;81370265;8147108;81400710);卫生部公益性研究基金(201192008);国家科技部十二五支撑计划(2012BAI10B01);黑龙江省中医药科研项目(ZHY12-Z190);佳木斯大学科学技术面上项目(L2012-049);黑龙江省卫生厅项目(2012-222)

摘　　要：目的探索骨髓间充质干细胞(Bone Marrow Mesenchymal Stem Cells,BMMSCs)和干细胞因子(Stem Cell Factor,SCF)促进肠黏膜细胞分化增殖和再生的机制,提供治疗老年炎症性肠病的认识。方法 (1)建立动物模型:由吲哚美辛诱导的胃肠黏膜损害,供体雄性SD老年大鼠皮下注射吲哚美辛处理(7.5mg/kg在5%碳酸氢钠)24小时连续3天,取4只吲哚美辛处理的SD老年大鼠使用戊巴比妥麻醉条件下处死(50mg/kg,腹腔注射)细胞组织学评估和确认老年炎症性肠病模型成功建立。(2)成功建立的老年炎症性肠病模型动物被分成4组,每组16只:BMMSCs-SCF治疗组,BMMSCs治疗组,SCF治疗组和生理盐水对照组。(3)使用PKH26荧光染色标记骨髓间充质基质细胞,输入炎症性肠病受试SD老年大鼠实验模型中,通过形态学观察测定肠黏膜的修复及再生,采用Western blot和PCR技术对动物模型体内骨髓间充质基质细胞中PKH26的表达情况。结果在受体老鼠胃肠黏膜中发现PKH26荧光染色标记的骨髓间充质基质细胞,如增殖细胞核抗原(proliferating cell nuclear antigen,PCNA),干细胞标记物(Lgr5,Musashi-1,ephrin-B3)等。在BMMSCs治疗组的SD老年大鼠肠黏膜中mRNA,PCNA的蛋白水平,Lgr5,Musashi-1和ephrin-B3增高,在BMMSCs-SCF混合治疗组中显著提高。在BMMSCs治疗组肠黏膜层和隐窝层浓度较高,更明显的在BMMSCs-SCF混合治疗组。结论在吲哚美辛诱导的肠道损伤实验中BMMSCs和SCF可能参与黏膜细胞再生并发挥协同作用。BMMSCs和SCF可能具有治疗价值。Objectives To explore the mesenchymal stromal cells and stem cell factor in the treatment of inflammatory bowel disease mechanism of bone marrow. Methods Use the PKH26 fluorescent dye labeled bone marrow mesenchymal stromal cells,input by indomethacin induced gastric mucosal damage of donor mice. The effects of bone marrow mesenchymal stromal cells transplantation were detected by the method of PKH26. The expression of PKH25 in bone marrow stromal cells in vivo was detected by blot Western and PCR. Results PKH26 fluorescent staining labeled MSCs were found in the gastrointestinal mucosa of rats, such as pro- liferating cell nuclear antigen （PCNA） ,stem cell marker （Lgr5）, Musashi-1 and tyrosine protein kinase B3 （ephrin-B3）. MRNA, LgrS, Musashi-1, ephrin-B3 and BMMSCs-SCF increased in the intestinal mucosa of the mice treated with bone marrow mesenchymal stromal cells. In the BMMSCs treatment group rats were significantly higher in the small intestine mucosal layer and the crypt layer, and more significantly in the BMMSCs-SCF mixed treatment group. Results：PKH26 fluorescent staining labeled MSCs were found in the gastrointestinal mucosa of rats, such as proliferating cell nuclear antigen （PCNA）, stem cell marker （LgrS）, Musashi-1 and tyrosine protein kinase B3 （ephrin-B3）. MRNA, LgrS, Musashi-1, ephrin-B3 and BMMSCs-SCF increased in the intestinal mucosa of the mice treated with bone marrow mesenchymal stromal cells. In the BMMSCs treatment group rats were significantly higher in the small intestine mucosal layer and the crypt layer, and more significantly in the BMMSCs-SCF mixed treatment group. Conclusion in the indomethacin induced intestinal injury in experimental bone marrow mesenchymal stromal cells and their soluble stem cell factor may participate in the mucosal cell regeneration and play a synergistic effect. Bone marrow stem cell therapy and stem cell factor management may be of therapeutic value.

关 键 词：髓间充质基质细胞 干细胞因子 炎症性肠病 

分 类 号：R574[医药卫生—消化系统]