摘要：

目的: 研究糖尿病对大鼠颌骨结构和颌骨来源骨髓间充质干细胞（bone marrow mesenchymal stem cells, BMSCs）成骨分化能力的影响，并初步探讨其可能的机制。方法: 采用链脲佐菌素腹腔注射建立大鼠糖尿病模型，正常对照组注射等量柠檬酸-柠檬酸钠缓冲液，每周监测血糖。建模成功后8周处死2组大鼠，微型CT（micro-CT）扫描重建上颌骨，测量牙槽骨吸收和颌骨骨形态学参数值。无菌条件下分离2组大鼠下颌骨，获取颌骨BMSCs。CCK-8法检测细胞增殖能力，碱性磷酸酶（alkaline phosphatase, ALP）染色和活性测定检测2组细胞ALP表达，茜素红染色和定量分析检测2组细胞钙结节形成能力，实时荧光定量PCR检测成骨相关基因Runx2和OCN的mRNA表达，western blot检测NF-κB p65核蛋白表达。结果: 与正常对照组相比，糖尿病组大鼠牙槽骨吸收增加（P<0.05），颌骨骨体积分数（bone volume/total volume，BV/TV）降低（P<0.05），骨小梁数目（trabecular number，Tb.N）减少（P<0.05），骨小梁间隙（trabecular space，Tb.Sp）增加（P<0.05）。成功分离、培养糖尿病大鼠颌骨来源的BMSCs，与正常大鼠颌骨BMSCs相比，细胞增殖能力降低（P<0.05），ALP活性和钙结节形成减少（P<0.05），成骨相关基因Runx2和OCN mRNA表达下降（P<0.05），NF-κB p65核蛋白表达增多（P<0.05）。结论: 糖尿病可影响大鼠颌骨结构，降低颌骨BMSCs成骨分化能力，机制可能与NF-κB信号通路激活相关。

关键词: 糖尿病, 颌骨骨髓间充质干细胞, 增殖, 成骨分化, NF-κB信号通路

Abstract:

Objective: To study the effect of diabetes on the jaw structure and the osteogenic differentiation ability of bone marrow mesenchymal stem cells (BMSCs) in rats and explore the possible mechanism. Methods: Diabetic rat models were established by injecting streptozotocin,which were used as the experimental group, rats receiving citric acid sodium citrate buffer injection were served as the control group. The alveolar bone resorption value of the maxilla and the microscopic morphological parameters of the maxilla were measured by micro-CT scanning. After culture of jaw BMSCs, cell proliferation activity were detected by CCK-8 test. Osteogenic differentiation activity was detected by alkaline phosphatase（ALP） test, alizarin red staining and quantitative analysis was used to detect calcium nodule formation. qRT-PCR was used to detect the mRNA expression of osteoblast-related genes Runx2 and OCN. Western blot was used to detect the expression of NF-κB p65 nuclear protein. Results: Micro-CT assay showed the alveolar bone absorption and trabecular space(Tb.Sp) increased, bone volume/total volume (BV/TV) and trabecular number (Tb. N) decreased in diabetic rats compared to normal rats (P<0.05). The BMSCs from jaw of diabetic rats were successfully isolated and cultured. Compared with the BMSCs from normal rats, the proliferation ability, ALP activity, calcium nodule formation and osteogenic gene expression of the diabetic rats BMSCs were all decreased (P<0.05), while the expression of NF-κB p65 nuclear protein was increased (P<0.05). Conclusion: Diabetes mellitus can affect the jaw structure of rats and reduce the osteogenic differentiation potentials of BMSCs. The mechanism may be related to the NF-κB signaling pathway.

Key words: diabetes mellitus, jaw bone marrow mesenchymal stem cells, proliferation, osteogenic differentiation, NF-κB signaling pathway

中图分类号: 

• R782