广西医学

目的观察不同浓度二甲双胍对大鼠离体成骨细胞增殖和分化的影响。方法从SD乳鼠颅骨中分离出原代成骨细胞后进行培养和鉴定。用不同浓度（0 μmol/L、50 μmol/L、100 μmol/L、200 μmol/L、400 μmol/L、800 μmol/L、1 600 μmol/L、3 200 μmol/L）二甲双胍干预成骨细胞72 h后，检测成骨细胞的增殖情况。用不同浓度（0 μmol/L、50 μmol/L、200 μmol/L、800 μmol/L、1 600 μmol/L、3 200 μmol/L）二甲双胍干预成骨细胞72 h后，检测成骨细胞的碱性磷酸酶（ALP）活性，以及骨形成指标ALP、Ⅰ型胶原（COL-Ⅰ）的基因和蛋白表达水平。结果（1）干预72 h后，与0 μmol/L二甲双胍相比，经50 μmol/L、100 μmol/L、200 μmol/L、400 μmol/L、800 μmol/L二甲双胍干预后成骨细胞的增殖活性呈浓度依赖性增强，其中经800 μmol/L二甲双胍干预的成骨细胞增殖活性最强（P＜0.05），而经3 200 μmol/L二甲双胍干预后成骨细胞的增殖活性降低（P＜0.05）。（2）在0～800 μmol/L浓度范围时，二甲双胍可浓度依赖性地增强成骨细胞中ALP的活性，但1 600 μmol/L及3 200 μmol/L二甲双胍可抑制成骨细胞中ALP的活性。（3）与0 μmol/L二甲双胍相比，经200 μmol/L、800 μmol/L二甲双胍干预后成骨细胞中ALP、COL-Ⅰ的mRNA和蛋白表达水平升高，而经3 200 μmol/L二甲双胍干预后成骨细胞中ALP、COL-Ⅰ的mRNA表达水平及COL-Ⅰ的蛋白表达水平下降（P＜0.05）；在0～3 200 μmol/L浓度范围内，二甲双胍浓度为800 μmol/L时成骨细胞中ALP、COL-Ⅰ的mRNA和蛋白表达水平最高（P＜0.05）。结论不同浓度二甲双胍对成骨细胞影响不同，较低浓度（≤800 μmol/L）二甲双胍可促进成骨细胞增殖和分化，较高浓度(3 200 μmol/L)二甲双胍则抑制成骨细胞增殖和分化。

ObjectiveTo observe the effect of metformin in different concentrations on proliferation and differentiation of isolated osteoblasts in rats. MethodsPrimary osteoblasts were isolated from skull of neonatal SD mice and cultured and identified. After 72 hours of intervention of osteoblasts with metformin in different concentrations (0 μmol/L, 50 μmol/L, 100 μmol/L, 200 μmol/L, 400 μmol/L, 800 μmol/L, 1600 μmol/L, and 3200 μmol/L), proliferation of osteoblasts was detected. After 72 hours of intervention of osteoblasts with metformin in different concentrations (0 μmol/L, 50 μmol/L, 200 μmol/L, 800 μmol/L, 1600 μmol/L, and 3200 μmol/L), alkaline phosphatase (ALP) activity of osteoblasts, and gene and protein expressions of bone morphogenesis indicator ALP and collagen type Ⅰ (COL-Ⅰ) were detected. Results(1) After 72 hours of intervention, compared with 0 μmol/L metformin, proliferation activity of osteoblasts was enhanced in a concentration-dependent manner after intervention with metformin of 50 μmol/L, 100 μmol/L, 200 μmol/L, 400 μmol/L, 800 μmol/L, therein, proliferation activity of osteoblasts with intervention of 800 μmol/L metformin was the strongest (P＜0.05), whereas proliferation activity of osteoblasts was decreased after intervention with 3200 μmol/L metformin (P＜0.05). (2) In the concentration range of 0-800 μmol/L, metformin could enhance ALP activity of osteoblasts in a concentration-dependent manner, but 1600 μmol/L and 3200 μmol/L metformin could inhibit ALP activity in osteoblasts. (3) Compared with 0 μmol/L metformin, mRNA and protein expressions of ALP and COL-Ⅰ in osteoblasts after intervention with 200 μmol/L and 800 μmol/L metformin were elevated, whereas mRNA expressions of ALP and COL-Ⅰ in osteoblasts, as well as protein expression of COL-Ⅰ in osteoblasts after intervention with 3200 μmol/L metformin were decreased (P＜0.05). In the concentration range of 0-3200 μmol/L, mRNA and protein expressions of ALP and COL-Ⅰ in osteoblasts were the highest when metformin was in concentration of 800 μmol/L (P＜0.05). ConclusionDifferent concentrations of metformin exert different effects on osteoblasts. Lower concentrations of metformin (≤800 μmol/L) can promote proliferation and differentiation of osteoblasts, while relatively higher concentrations of metformin (3200 μmol/L) can inhibit proliferation and differentiation of osteoblasts.