| 褚亚男,鲍鹏丽,马士凤,王 静,周 娜,郑荣秀.尾静脉注射骨髓间充质干细胞对1型糖尿病幼鼠的治疗作用实验研究[J].现代生物医学进展英文版,2021,(6):1008-1013. |
| 尾静脉注射骨髓间充质干细胞对1型糖尿病幼鼠的治疗作用实验研究 |
| Experimental Study on the Therapeutic Effect of Tail Vein Injection of Bone Marrow Mesenchymal Stem Cells in Type 1 Diabetic Infant Rats |
| Received:August 28, 2020 Revised:September 23, 2020 |
| DOI：10.13241/j.cnki.pmb.2021.06.002 |
| 中文关键词: I型糖尿病 链脲佐菌素 骨髓间充质干细胞 |
| 英文关键词: Type 1 Diabetes Streptozotocin Bone marrow Mesenchymal Stem Cells |
| 基金项目:天津市自然科学基金项目（17JCZDJC36400）；天津市卫生局科研基金项目（16KG123） |
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| 中文摘要: |
| 摘要 目的：比较尾静脉注射骨髓间充质干细胞（BMSCs）治疗1型糖尿病（type 1 diabetes,T1D）幼鼠的效果。方法：选用3周龄C57bl/c幼鼠作为受试动物，连续5 d腹腔注射50 mg/kg的链脲佐菌素（STZ），建立T1D模型；采用酶消化法联合骨片法从2周龄C57bl/c幼鼠的胫骨和股骨中分离出BMSCs；碱性磷酸酶（ALP）和油红O染色检测P3代BMSCs的诱导分化能力；流式细胞仪鉴定P3代BMSCs的细胞表型；采用生理盐水和不同剂量的BMSCs（低剂量6×105 cells/mL、中剂量1.2×106 cells/mL和高剂量2.4×106 cells/mL）通过尾静脉输注的方式对T1D幼鼠进行治疗，定期检测T1D幼鼠的体重、血糖变化；T1D幼鼠治疗28 d后，取其胰脏行病理学分析。结果：（1）3周龄C57bl/c幼鼠注射STZ后14 d，幼鼠表现为体重增长缓慢、血糖明显升高；（2）分离得到的BMSCs细胞呈长梭纤维状；BMSCs成骨诱导9 d，碱性磷酸酶（ALP）染色后细胞外基质有大量碱性磷酸酶表达；BMSCs成脂诱导14 d，油红O染色后细胞内有大量脂滴出现；流式细胞仪检测BMSCs细胞表型，BMSCs不表达CD31、CD34和CD45，高表达CD29、CD90和CD105；（3）T1D幼鼠经过BMSCs治疗后，其体内的血糖下降并保持稳定；H.E.和胰岛素免疫组织化学染色结果显示实验组T1D幼鼠的胰腺组织随着治疗时间的延长，其损伤的胰腺组织得到了逐步的恢复，而未经过任何治疗的T1D幼鼠，其胰腺组织的损伤在逐步加重。结论：尾静脉注射BMSCs对于T1D幼鼠有治疗效果。 |
| 英文摘要: |
| ABSTRACT Objective: To compare the effect of tail vein injection of bone marrow mesenchymal stem cell on type 1 diabetes(T1D) in immature C56bl/c mice. Methods: 3 weeks old C56bl/c mice was injected streptozotocin(STZ) 50 mg/kg by intraperitoneal for 5 days. BMSCs were isolated from Tibia and Femur of C56bl/c mice by enzyme digestion and bone slice method. The induced differetiation ability of the BMSCs was detected by alkaline phosphatase staining and Oil-red-O staining. The flow cytometry was used to detect the phenotypes of BMSCs. The T1D mice were treated with normal saline and different doses of BMSCs by intravenous infusion, and the body weight and blood glucose of T1D mice were detected regularly. The pancreas of the T1D mice was taken for pathological analysis which were treated by BMSCs for 28 days later. Results: (1)The 3-weeks-old C56bl/c mice showed slow weight gain and significantly blood glucose 14 days after the STZ injection. (2) The BMSCs were exhibited long spindle like fibers. ALP and Oil-red-O staining showed that BMSCs could be induced to Osteoblast and adipocyte. Flow cytometry analysis showed that BMSCs overexpressed CD29, CD90 and CD105, and rarely expressed CD31, CD34 and CD45. (3) The blood glucose of T1D mice decreased and remained stable which treated with BMSCs. Pathological examinations on pancreas islets by H.E. staining and insulin immunohistochemistry demonstrated that the pancreas islet of T1D mice in MSC treatment groups were recovered with the time of treatment, while the without any treatment group of the pancreas islet was getting worse. Conclusion: BMSCs had a significant effect on T1D immature mice. |
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