人端粒酶反转录酶转染的骨髓间充质干细胞移植治疗大鼠糖尿病

Human telomerase reverse transcriptase transfection of bone marrow mesenchymal stem cells in the treatment of diabetic rats

背景:人端粒酶反转录酶是调控增殖及定向分化的首选生长因子之一,具有多重生物学效应。目的:观察人端粒酶反转录酶表达的骨髓间充质干细胞移植治疗大鼠糖尿病的效果。方法:体外培养SD大鼠骨髓间充质干细胞,经反转录病毒PLXSN为载体介导人端粒酶反转录酶基因转染骨髓间充质干细胞,在转染前后用RT-PCR检测骨髓间充质干细胞人端粒酶反转录酶基因的表达。60只雌性SD大鼠中随机取15只作为正常对照组,一次性注射生理盐水,余45只按45 mg/kg的剂量注射链脲霉素建立糖尿病模型后,随机分为3组,分别通过大鼠尾静脉注射移植人端粒酶反转录酶基因转染的骨髓间充质干细胞0.2 mL、骨髓间充质干细胞0.2 mL、生理盐水0.2 mL。结果与结论:转染48 h后发现,骨髓间充质干细胞有人端粒酶反转录酶mR NA的表达,且重点集中于胞核内。移植后14 d,糖尿病组大鼠空腹血糖维持在较高水平,且高于正常对照组(P<0.05);与糖尿病组相比,各移植组大鼠空腹血糖水平显著下降(P<0.05);与骨髓间充质干细胞移植组相比,人端粒酶反转录酶基因转染的骨髓间充质干细胞移植组大鼠空腹血糖水平显著下降(P<0.05),接近正常对照组水平(P>0.05)。结果提示人端粒酶反转录酶表达的骨髓间充质干细胞移植能有效治疗大鼠糖尿病。

BACKGROUND: Human telomerase reverse transcriptase(h TERT) is the first choice for regulating the proliferation and directional differentiation, with multiple biological effects.OBJECTIVE: To observe the therapeutic effect of h TERT-transfected bone marrow mesenchymal stem cells transplantation in diabetic rats.METHODS: Bone marrow mesenchymal stem cells from Sprague-Dawley rats were cultured in vitro and transfected with retrovirus PLXSN carrying hT ERT. RT-PCR was used to detect the hT ERT expression in the bone marrow mesenchymal stem cells before and after transfection. Sixty male Sprague-Dawley rats were selected and equally randomized into four groups: normal control group, transfection group, cell transplantation group, and model group. In the latter three groups, rats were injected with 45 mg/kg chain urea to establish diabetes models,and then injected via the tail vein with 0.2 mL hT ERT-transfected bone marrow mesenchymal stem cells, 0.2 m L bone marrow mesenchymal stem cells, and 0.2 m L normal saline, respectively.RESULTS AND CONCLUSION: At 48 hours after hT ERT transfection, the expression of hT ERT m RNA was detected in the bone marrow mesenchymal stem cells, and mainly concentrated in the nuclei. At 14 days after transfection, the fasting glucose level in the model group was higher than that in the normal control group(P < 0.05). However, compared with the model group, the fasting glucose levels in the transfection and cell transplantation groups were significantly decreased(P < 0.05); compared with the cell transplantation group, the fasting glucose level in the transfection group was also significantly decreased(P < 0.05), which was similar to thatin the normal control group(P > 0.05). These findings indicate that the transplantation of hT ERT-transfected bone marrow mesenchymal stem cells is effective in the treatment of diabetic rats.