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目的骨髓间充质干细胞(BMSCs)移植入糖尿病模型鼠胰腺包膜下,观察BMSCs的分布及对糖尿病的治疗效果。方法利用绿色荧光蛋白(EGFP)标记雄性SD大鼠BMSCs,以多点注射的方法移植入糖尿病大鼠胰腺包膜下,血糖仪监测血糖,ELISA法检测胰岛素和C-肽水平。移植后8周,通过EGFP示踪植入的BMSCs的分布,取EGFP标记的胰腺组织,免疫荧光染色检测是否存在EGFP和胰岛素共表达,荧光原位杂交检测是否存在EGFP和PDX1共表达。结果胰腺包膜下移植BMSCs有效降低糖尿病鼠血糖,升高血清胰岛素和C-肽水平(P<0.05)。至移植后8周,植入细胞稳定表达绿色荧光蛋白,EGFP标记细胞分布于胰岛(12.46%)、血管(4.4%)、腺泡(9.24%)、导管(3.21%),或集中分布于局部(70.69%)。免疫组化发现EGFP和胰岛素共表达细胞(5.16%),荧光原位杂交发现EGFP和PDX1共表达细胞(0.96%)。结论BMSCs胰腺包膜下移植后在胰腺中发生再分布,在胰腺微环境中能够分化为胰岛素分泌细胞,有效逆转血糖、胰岛素和C-肽水平。
Objective Bone marrow mesenchymal stem cells (BMSCs) were transplanted into the pancreas of diabetic model rats to observe the distribution of BMSCs and the therapeutic effect on diabetes mellitus. Methods Male Sprague-Dawley rat BMSCs were labeled with green fluorescent protein (EGFP) and injected into the pancreas of diabetic rats by multi-point injection. The blood glucose was measured by blood glucose meter. The levels of insulin and C-peptide were measured by ELISA. Eight weeks after transplantation, the distribution of BMSCs implanted by EGFP was detected. The EGFP-labeled pancreatic tissues were taken and immunofluorescent staining was used to detect the presence of EGFP and insulin co-expression. The co-expression of EGFP and PDX1 was detected by fluorescence in situ hybridization. Results BMSCs transplanted into the pancreas effectively reduced the blood glucose, increased the levels of serum insulin and C-peptide in diabetic rats (P <0.05). At 8 weeks after transplantation, EGFP was stably expressed in the implanted cells. The EGFP-labeled cells were located in islets (12.46%), blood vessels (4.4%), acinar (9.24%), catheters (70.69%). EGFP and insulin co-expressing cells were found by immunohistochemistry (5.16%). EGFP and PDX1 co-expressing cells (0.96%) were found by fluorescence in situ hybridization. CONCLUSION: BMSCs can be redistributed in the pancreas under the pancreas and can differentiate into insulin-secreting cells in the pancreas microenvironment, effectively reversing the levels of blood glucose, insulin and C-peptide.