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目的研究骨形态发生蛋白7(BMP7)对体外培养高磷诱导的大鼠血管平滑肌细胞(VSMC)钙化的影响。方法应用大鼠主动脉VSMC进行体外实验,分为3组:正常组(磷浓度为1.4mmol/L,n=6)、高磷组(磷浓度为2.4mmol/L,n=6)和高磷+BMP7(200μg/L)组(n=6),分别培养3、6、9d。应用甲σ酚肽络合酮法检测不同时间细胞内的钙含量,放射免疫方法检测培养骨钙素含量,ELISA检测不同时间的核结合因子α1(Cbfα1)与DNA结合的活性,免疫组织化学方法检测α平滑肌肌动蛋白(α-SMA)的表达。结果与正常组和高磷+BMP7组比较,高磷组VSMC钙含量、上清液骨钙素含量[(14.62±2.93)比(2.83±0.64)、(9.36±1.37)μg/g蛋白]、Cbfα1活性均明显增加(均P<0.05);与高磷组比较,高磷+BMP7组的α-SMA阳性细胞百分比显著增多(P<0.05)。结论高磷可直接刺激大鼠VSMC的钙沉积和增加骨钙素,其作用可能与高磷上调Cbfα1表达有关;BMP7可抑制大鼠VSMC的钙沉积和骨钙素产生,作用可能与其降低Cbfα1表达有关。
Objective To investigate the effect of bone morphogenetic protein 7 (BMP7) on the calcification of rat vascular smooth muscle cells (VSMCs) induced by high phosphorus in vitro. Methods The rat aorta VSMCs were divided into three groups: normal group (1.4 mmol / L phosphorus concentration, n = 6), high phosphorus group (2.4 mmol / L phosphorus concentration, n = 6) Phosphorus + BMP7 (200μg / L) group (n = 6) were cultured for 3,6,9d. The content of calcium in cells at different time was detected by alpha-phenol-phenol-complexed ketone method, the content of osteocalcin was detected by radioimmunoassay, the activity of DNA-binding activity of nuclear factor-α1 (Cbfα1) at different time was detected by ELISA, The expression of α-smooth muscle actin (α-SMA) was detected. Results Compared with the normal group and the high phosphorus + BMP7 group, the content of calcium in high phosphorus group, the content of osteocalcin in supernatant [(14.62 ± 2.93) vs (2.83 ± 0.64), (9.36 ± 1.37) μg / g protein] (All P <0.05). Compared with high phosphorus group, the percentage of α-SMA positive cells in high phosphorus + BMP7 group increased significantly (P <0.05). Conclusion High phosphorus can directly stimulate calcium deposition and increase osteocalcin in rat VSMCs, which may be related to the up-regulation of Cbfα1 expression by high phosphorus levels. BMP7 can inhibit calcium deposition and osteocalcin production in VSMCs of rats, which may be related to the decrease of Cbfα1 expression related.