大鼠成体干细胞食管下段移植的实验研究

Study on transplantation of adult stem cells to rat lower esophagus: a potential treatment for gastroesophageal reflux disease

目的研究成体干细胞注射至食管下段的可行性,探索利用成体干细胞[骨骼肌卫星细胞(SC)和骨髓间充质干细胞(MSC)]移植治疗胃食管反流病(GERD))的新方法。方法选取体质量 160～240 g的Wistar大鼠,雌雄各半,不同实验组分别于体外培养并扩增骨骼肌SC或MSC, 5-溴-2-脱氧尿嘧啶核苷(BrdU)标记后,经腹腔将大鼠自体骨骼肌SC悬液植入食管下段肌层;MSC悬液植入同种异体大鼠食管下段肌层。1周和4周后分批处死动物,取食管下段组织并使用组织学方法评价植入细胞生长分化情况。结果 BrdU的细胞标记率接近100％。接受干细胞移植的大鼠生长良好。干细胞移植后第1周和第4周,SC组和MSC组大鼠食管下段均可见大量分化为骨骼肌细胞的 BrdU阳性细胞,SC组第4周较第1周BrdU阳性细胞核数显著增多(P<0．01)。MSC组第4周较第1周 BrdU阳性细胞核计数增多,但差异无统计学意义(P>0．05)。SC组与MSC组比较,第1周BrdU阳性细胞核数差异无统计学意义(P>0．05),第4周SC组较MSC组显著增多(P<0．05)。注射局部未见炎症反应,与对照组病理学分析无差异。结论成体干细胞移植至大鼠食管下段安全、可行,能在食管下段长期存活并分化为骨骼肌。骨骼肌SC或MSC移植可能成为治疗GERD的新方法。

Objective To explore the feasibility and safety of the transplantation of adult stem cells, skeletal muscle satellite cells （SMSC） and bone marrow mesenchyrnal stem cells （BMMSC）, into lower esophagus of rats as a new potential method to treat gastroesophageal reflux disease. Methods SMSC and BMMSC were procured from tibialis anterior muscles or the whole hind limbs of adult Wistar rats respectively, which were cultured, proliferated and labelled with 5-bromo-2-deoxyuridine （BrdU） in vitro. SMSCs were autologously implanted into the lamina muscularis of lower esophagus, and BMMSCs were injected into the same position of allogenic rats. The esophagus tissue was harvested 1 and 4 weeks later for histological study. Resets The labelling efficiency of the ceils with BrdU was nearly 100%. One and four weeks after cell transplantation, a lot of BrdU positive cells were observed in the lamina muscularis of lower esophagus in both of the groups, which had differentiated into esophageal skeletal muscle cells. In SMSC group, there were more BrdU-labelled cells at the fourth week than those at the first week（P〈0. 01）, but there was only a slight increase of BrdU-labelled cells at the fourth week as compared with the first week in BMMSC group（P〉0.05）. However, there were no significant differences in SMSC and BMMSC groups at the first week（P〉0. 05）. No obvious local inflammation was found. Conclusions The study proved the feasibility and persistence of injecting SMSC and BMMSC into the lower esophagus. Transplantation of adult stem cells may serve as a potential new treatment for gastroesophageal reflux disease.