摘要
目的:探讨骨髓间充质干细胞在胰腺生理更新和病理再生中的作用. 方法:将60只SD大鼠随机分为5组,每组12 只．A组为正常阴性对照组,不作任何处理,B 组为自体骨髓回输对照组,C组为雨蛙肽致轻度胰腺炎(MAP)模型组,D组为L-精氨酸致重度胰腺炎(SAP)模型组,E组为粒细胞集落因子(G-CSF)预处理组(SAP+GSF):在造模前3 d,sc G-CSF 40μg(kg·d),共3 d,其余处理同 D组．造模前3 d用核染料Hoechst33258标记自体骨髓间充质干细胞(MSC),并回输到自体骨髓腔．于骨髓干细胞回输后的2 wk和8 wk,分批处死大鼠,采集胰腺,立即进行冰冻切片, 荧光显微镜下直接观察胰腺组织中是否有呈现黄绿色荧光的细胞,选择有上述黄绿色荧光的冰冻切片进行免疫荧光染色,观察有无 Cytokeratin 19、胰岛素和胰高血糖素染色呈阳性的细胞．结果:A组的胰腺冰冻切片未见有黄绿色荧光．造模后2 wk,B、C、E和D组存活大鼠可见标记的MSC出现在正常胰腺组织和损伤胰腺组织中,在正常胰腺组织中偶见,而在损伤胰腺组织中多见,尤以GSF预处理组最多见, C、D、E组一直持续到8 wk仍可见到黄绿色荧光．A和D组死亡大鼠未见Cytokeratin 19、胰岛素和胰高血糖素染色呈阳性的细胞．造模后2 wk,B、C、E和D组存活大鼠掺入损伤胰腺中的MSC均有分化为Cytokeratin 19染色阳性的细胞,未见胰岛素和胰高血糖素染色呈阳性的细胞．造模后8 wk,胰岛素和胰高血糖素染色呈阳性的细胞在C、D组偶见,E组散见．结论:自体骨髓间充质干细胞参与胰腺的生理更新和病理再生．
AIM： To investigate the role of bone marrow mesenchymal stem cells （MSC） in pancreatic self-restoration and pathological regeneration. METHODS： Sixty Sprague-Dawley （SD） rats were randomly assigned into 5 groups （n =12）. Group A acted as the normal negative control without any treatment, and group B received labeled autologous bone marrow MSC only. Group C was induced mild acute pancreatitis by subcutaneous injection of caerulein 20 μg/ （kg·h） 4 times, and group D was induced severe acute pancreatitis by intraperitoneal injection of L-arginine （2 g/kg） twice. Group E was the stem cell mobilized group treated by injection of granulocyte-colony stimulating factor （G-CSF） into rats for 3 d at a dose of 40 μg/（kg· d） 3 d before the induction of SAP. MSC were stained with Hoechst33258 and transplanted into their original cavity. Two and eight weeks after transplantation, the rats were sacrificed, and pancreatic tissues were harvested. Samples were snap-frozen and sectioned on a cryostat. The presence of labeled MSC in the cryostat prepared was examined directly by fluorescent microscopy. The positive sections were selected for further immunofluorescence assay. Anti-Cytokeratin（CK）19, anti-glucagon and antiinsulin immunofluorescence staining were performed on the pancreatic sections to determine whether incorporated MSC differentiated into mature pancreatic cells. RESULTS： Frozen section of pancreas in group A didn＇t appear yellow-green fluorescence. The labeled MSC were detected in normal pancreatic tissues in group B and in injured pancreatic tissues in group C, D and E, especially in group E, which lasted 8 wk. The results of immunofluorescence analysis were as follows： All tests were negative in controls, including spontaneous fluorescence control, fluorescence antibody control and inhibitory control. No positive cells with CK19, insulin and glucagons were visualized in group A and D （died rats）. Positive cells with CK19, which differentiated from the labeled MSC, were found in normal pancreas in group B and injured pancreatic models in group C, E and D （survival rats）. No positive cells with insulin and glucagons were observed 2 wk after modeling, while the positive cells appeared 8 wk after modeling in group C, D and E. CONCLUSION： The bone marrow MSC participate in pancreatic self-restoration and pathological regeneration.
出处
《世界华人消化杂志》
CAS
北大核心
2006年第4期398-404,共7页
World Chinese Journal of Digestology
关键词
骨髓间充质干细胞
胰腺干细胞
胰腺炎
Bone marrow mesenchymal stem cells
Acute pancreatitis
Pancreatic stem cells
