黄芪调控体外培养大鼠成骨细胞Ⅰ型胶原蛋白的表达

Astragalus root injection regulates type Ⅰ collagen expression of rat osteoblasts in vitro

背景:Ⅰ型胶原是体外培养成骨细胞所分泌的特异性胶原蛋白,所形成的网状结构也是成骨细胞矿化功能实现的前提,也是反映成骨细胞骨形成能力的重要指标。诸多报道反映了应用单味黄芪促进成骨细胞增殖的功能,但缺少有关黄芪促进Ⅰ型胶原分泌及表达的研究。目的:观察黄芪注射液对体外培养新生大鼠颅盖骨成骨细胞增殖及分泌Ⅰ型胶原能力的影响。方法:体外分离培养大鼠成骨细胞。分为用含小牛血清的MEM培养液培养的对照组,分别加入相应体积分数黄芪注射液培养的黄芪组,于培养1,3,5,7,9d以四甲基偶氮唑盐法观察成骨细胞增殖的影响;以Real-timePCR方法检测黄芪注射液干预6h后成骨细胞Ⅰ型胶原蛋白α1mRNA的表达情况,并用incellwestern方法测定培养6d时成骨细胞Ⅰ型胶原蛋白的分泌。结果与结论:从培养3d开始直至培养9d,与对照组比较,各体积分数的黄芪注射液均能促进成骨细胞的增殖(P<0.05),这一趋势在培养7d时达到最高峰。不同体积分数的黄芪注射液均能促进COLLα1mRNA的表达,尤其以15%的黄芪作用明显。以15%和10%的黄芪注射液培养的成骨细胞Ⅰ型胶原的表达量明显高于对照组(P<0.05)。结果证实,黄芪可以促进体外培养的成骨细胞的增殖和分泌Ⅰ型胶原的能力,但具有一定的量效趋势。

BACKGROUND： Type Ⅰ collagen is a specific collagen secreted by in vitro cultured osteoblast, and the formed network is the basis of bone mineralization, which also reflects the ability of osteoblast bone formation. Studies have shown astragalus root increased osteoblast proliferation. However, the effect of astragalus root on improving type Ⅰ collagen expression of osteoblast remains poorly understood. OBJECTIVE： To evaluate the effect of astragalus root injection on the abilities of rat cranium-derived osteoblast proliferation and type Ⅰ collagen expression. METHODS： Rat osteoblast was cultured in vitro and divided into control group （MEM culture solution containing calf serum） and astragalus root groups （different concentrations）. The effect on osteoblast proliferation was evaluated on days 1, 3, 5, 7, and 9 by MTT method. Moreover, the expression of type Ⅰ collagen protein was observed after 6 hours of treatment with astragalus root injection using in cell western-blot method. In addition, the gene expression of COLLα1 was investigated by real-time PCR method. RESULTS AND CONCLUSION： From days 3 to 9, the different concentrations of astragalus root injection improved osteoblast proliferation, respectively compared with control group （P 0.05）, and this ascending trend peaked on day 7. Different concentrations of astragalus root injection improved COLLα1 mRNA expression, especially 15% astragalus root injection was the most effective. The type Ⅰ collagen protein expression of 15% and 10% astragalus root injection were significantly greater compared with the control group （P 0.05）. Astragalus root injection improved in vitro cultured osteoblast proliferation and type Ⅰ collagen secretion in a certain dose-effect manner.