骨桥蛋白干预体外培养人膝骨关节炎软骨细胞蛋白多糖和Ⅱ型胶原的表达

Osteopontin promotes the expression of aggrecan and type Ⅱ collagen in osteoarthritic chondrocytes of the knee joint in vitro

背景:骨桥蛋白基因和蛋白表达与骨关节炎的病变程度高度相关。目的:进一步验证骨桥蛋白对体外培养人膝骨关节炎软骨细胞蛋白多糖和Ⅱ型胶原mR NA表达的影响。方法:体外培养人膝骨关节炎软骨细胞。取第1代人膝骨关节炎软骨细胞,分为空白对照组、0.1 mg/L骨桥蛋白组和1 mg/L骨桥蛋白组。骨桥蛋白组加入重组骨桥蛋白0.1 mg/L或1 mg/L干预48 h;空白对照组未予任何干预继续培养48 h。用Real-time PCR测量蛋白多糖与Ⅱ型胶原的mR NA表达量。结果与结论:经0.1 mg/L与1 mg/L两种质量浓度骨桥蛋白干预后,人膝骨关节炎软骨细胞蛋白多糖与Ⅱ型胶原mR NA表达水平显著上升(P<0.05),1 mg/L骨桥蛋白组人膝骨关节炎软骨细胞蛋白多糖与Ⅱ型胶原mR NA表达水平高于0.1 mg/L骨桥蛋白组(P<0.05),人膝骨关节炎软骨细胞蛋白多糖mR NA表达水平与骨桥蛋白干预质量浓度呈正相关(r=0.751,P<0.01)。人膝骨关节炎软骨细胞Ⅱ型胶原mR NA表达水平与骨桥蛋白干预浓度呈正相关(r=0.676,P<0.01)。结果提示骨桥蛋白上调人膝骨关节炎软骨细胞蛋白多糖与Ⅱ型胶原mR NA的表达,并具有浓度依赖性。

BACKGROUND： Osteopontin m RNA and protein expressions are highly correlated with the severity of osteoarthritis. OBJECTIVE： To investigate the effect of osteopontin on the gene expression of aggrecan and type Ⅱ collagen inthe human knee osteoarthritic chondrocytes in vitro. METHODS： Chondrocytes were harvested from human osteoarthritic knees and cultured in vitro. The chondrocytes were cultured with 0（blank control group）, 0.1, 1 mg/L osteopontin, respectively, for 48 hours. Real-time PCR was employed to detect the m RNA expression of aggrecan and type Ⅱ collagen. RESULTS AND CONCLUSION： After 0.1 and 1 mg/L osteopontin intervention, the m RNA expression of aggrecan and type Ⅱ collagen in osteoarthritic chondrocytes was increased significantly（P〈0.05）, and the m RNA expression of aggrecan and type Ⅱ collagen was higher in the 1 mg/L osteopontin group than the 0.1 mg/L osteopontin group（P〈0.05）. In addition, the m RNA expression of aggrecan and type Ⅱ collagen was positively correlated with the concentration of osteopontin（r=0.751, P〈0.01; r=0.676, P〈0.01）. These findings indicate that osteopontin up-regulates the m RNA expression of aggrecan and type Ⅱ collagen in osteoarthritic chondrocytes of human knee in vitro in a dose-dependent manner.