骨髓间充质干细胞自体介入移植治疗糖尿病Beagle犬

Reversal of diabetes in Beagle dogs by intrahepatic autotransplantation of bone-derived mesenchymal stem cells

目的 观察转人胰岛素（Insulin,INS）和绿色荧光蛋白（EGFP）基因的Beagle犬骨髓间充质干细胞（BcMSCs）经动脉介入移植入糖尿病Beagle犬自体肝内后的长时程疗效及安全性.方法 构建逆转录病毒双表达载体pMSCV-INS-IRES-EGFP,转染BcMSCs后,经肝动脉介入移植到糖尿病Beagle犬自体肝脏.结果 糖尿病Beagle犬治疗组接受移植后,体质量至第10周上升8.83%.移植后第7、70天空腹血糖值分别为（19.80±3.41）、（10.53±2.12）mmol/L,与对照组差异有统计学意义（P＜0.05）.血清及肝脏中检测到人胰岛素.静脉葡萄糖耐量实验示血糖水平下降.形态学分析证实移植入的BcMSCs在肝内定植并分泌人胰岛素,移植部位无病理形态学改变.结论 该法能有效降低糖尿病Beagle犬血糖水平.

Objective To assess the long-time curative effect and safety of intrahepatic autotransplantation of bone-derived Beagle canine mesenchymal stem cells （BcMSCs） containing the human Insulin （INS） and enhanced green fluorescent protein （EGFP） gene through artery intervention in diabetic Beagle dogs. Methods After the double expression vector pMSCV-INS-IRES-EGFP was constructed, BcMSCs were isolated from the bone marrow of Beagle canine, expanded, and transfected with viral suspension.Then the cells were treated with G418 and cultured in vitro, followed by intrahepatic autotransplantation to the diabetic Beagle canine. Results The body weight of diabetic Beagle dogs that received BcMSCs harboring the human INS and EGFP gene stopped dropping and increased by 8.83% within 10 weeks after treatment, and the average blood glucose levels were （ 19. 80 ±3. 41 ） mmol/L （at day 7） and （ 10. 53 ±2. 12）mmol/L （ at day 70） respectively. There was significant difference in the average blood glucose level between BcMSCs group and control group （P 〈 0. 05）. The human INS was detected in blood serum and the liver of the treated animals. Intravenous glucose tolerance test revealed that in BcMSCs group the blood glucose level was reduced. Moreover, pathomorphological research confirmed that the transplanted BcMSCs were permanently planted in the animal liver and could secrete human INS. No pathomorphological changes were found in the transplant part. Conclusion Intrahepatic autotransplantation of BcMSCs expressing human INS could effectively reduce the blood glucose level and alleviate the symptoms of diabetes.