摘要
目的观察植入糖尿病大鼠体内的骨髓间充质干细胞(MSCs)能否分化为可分泌胰岛素的细胞或者促进胰岛β细胞增殖。方法采用贴壁法分离培养来自同种异体大鼠的MSCs,移植前用5-溴-2-脱氧尿嘧啶核苷(BrdU)标记。将60只雄性SD大鼠随机分为正常对照组(NC组);糖尿病大鼠对照组(DM组);经左心腔注射移植MSCs糖尿病大鼠组(MSCs组);经左心腔注射移植MSCs并行环孢霉素A灌胃的糖尿病大鼠组(MSCs+CsA组)。分别于造模前、成摸时和移植细胞后7、14、28天时检测OGTT和体重;每周固定时间测一次随机血糖;处死大鼠前从心脏取血测胰岛素;移植细胞后7、14和28天时,取各组大鼠胰腺组织行增殖细胞核抗原和Brdu免疫组化。结果移植MSCs后7天和14天都可在MSCs组和MSCs+CsA组大鼠胰腺的血管内及其周围、胰腺外分泌组织和/或胰岛内发现Brdu阳性细胞。移植MSCs后14天MSCs+CsA组OGTT的2h血糖值较DM组有下降趋势(P=0.069);三组糖尿病大鼠间胰岛素水平和体重无明显差异;移植MSCs未使MSCs和MSCs+CsA组胰岛中增殖细胞明显增多。结论通过左心室注入的大鼠MSCs可以迁移到损伤的胰腺组织中,植入的MSCs使糖尿病大鼠的血糖下降不显著,不能证明能分化为可分泌胰岛素的细胞,也未促进胰岛细胞增殖;在胰岛损伤高峰期移植细胞,很难达到治疗的目的。
Objective To evaluate the potential capacity for the intracardially transplanted allogeneic bone marrow mesenchymal stem cells （MSCs） to differentiate into insulin-secreting cells or to improve the proliferation of pancreatic islets. Methods MSCs were isolated from bone marrow by adhesive screening method, cultured and marked with 5-bromo2＇-deoxyuridine （grdu） in vitro. The male SD rats were randomly allocated to normal control group （Group A）, STZ-induced diabetic rats without treatment （Group B）, MSC-treated diabetic rats （Group C） and MSC-treated diabetic rats with intragastric administration of cyclosporine A （CsA） （Group D）. Oral glucose tolerance tests （OGTT） were carried out before STZ injection, at about 10 days after STZ injection and at the 1^st, 2^nd or 4^th week after transplantation of MSCs by the intracardial injection. At the 1^st, 2^nd or 4^th week after transplantation of MSCs, the pancreatic tissues of rats were removed for immunohistochemistry staining of BrdU and proliferating cell nuclear antigen （PCNA）. And then the distribution of BrdU-positive cells in pancreas was observed and the quantity of PCNA-positive cells in pancreatic islets was evaluated. Results At the 1^st or 2^nd weeks after transplantation of MSCs, the BrdU-positive cells were easily observed in the blood vesseles, excrine tissues of pancreas and pancreatic islets. At thesecond week after transplantation of MSCs, the rats of group D had a trend toward blood glucose decrease at 120 rain compared with that of group B（P=0. 069）. After transplantation of MSCs, group C and D did not show obviously higher BW and insulin levels （P 〉 0.05） compared with group B. The transplanted MSCs did not improve the proliferation of islet ceils in group C and D as compared with group B （P〉0. 05）. Conclusions The MSCs by the intracardial injection could engraft into injured pancreas tissues and may induce a slight decrease of blood glucose, but the cells could not secret insulin and not improve the proliferation of islet cells. The mechanisms need to be further investigated. During the severe injury of pancreatic islet, it is difficult for transplanted MSCs to restore islet cells.
出处
《中国糖尿病杂志》
CAS
CSCD
北大核心
2009年第7期510-515,共6页
Chinese Journal of Diabetes
