骨保护素参与高磷诱导大鼠血管平滑肌细胞钙化调节的机制初探 #br#

摘要：目的 以β-磷酸甘油诱导大鼠血管平滑肌细胞（VSMC）钙化模型为研究对象，初步探讨骨保护素（OPG）干 预对VSMC钙化的影响及机制。方法 体外分离培养大鼠主动脉VSMC。PBS为对照组，以10 mmol/L β-磷酸甘油诱 导钙化10 d为钙化对照组，β-磷酸甘油诱导钙化同时予以剂量梯度（1、4、8 μg/L）的OPG干预为OPG干预组。茜素红 染色检测各组钙化结节形成情况。邻甲酚酞络合酮比色法检测细胞钙沉积含量。比色法测定碱性磷酸酶（ALP）活性。 免疫蛋白印迹（Western blot）检测各组Notch1/RBP-JK通路蛋白的表达。结果 茜素红染色显示，与钙化对照组比较， OPG干预组大鼠VSMC钙化结节明显减少，OPG干预组的钙含量、ALP活性均明显下降（均P＜0.05），且有剂量依赖性。 Western blot结果显示，与钙化对照组比较，OPG干预组的Notch1、RBP-JK、Jagged1、Msx2蛋白表达水平明显下降（均P＜ 0.05）。结论 OPG可能通过下调Notch1/RBP-JK通路蛋白表达，参与大鼠VSMC钙化过程调节。

The mechanism of osteoprotegerin involved in the calcification of vascular smooth muscle cells induced by β-glycerophosphate #br#

Abstract：Objective To investigate the mechanism of osteoprotegerin intervention on thecalcification of vascular smooth muscle cells (VSMC) induced by β -glycerophosphate in rats. Methods Rat aortic smooth muscle cells were isolated and cultured in vitro. Cells treated with PBS were used as the control group. Cells treated with 10 mmol/L β - glycerophosphate for 10 days to induce the calcification were used as the calcified control group, while cells treated with β- glycerophosphate and osteoprotegerin (OPG) in dose gradients (1, 4 and 8 μg/L) were used as the osteoprotegerin intervention group. Alizarin red staining was used to detect the formation of calcified nodules. The content of calcium deposition was measured by the o-cresolphthalein complexone method. The alkaline phosphatase activity was determined by the colorimetric method. The protein expressions of Notch1/RBP-JK pathway were detected by Western blot assay. Results Alizarin red staining showed that compared with the calcified control group, the calcified nodules of VSMCs were significantly reduced in OPG intervention groups. Compared with calcification control group, the calcium content and ALP activity decreased significantly in OPG intervention group in a dose-dependent manner (P < 0.05). In addition, compared with calcification control group, the expression levels of Notch 1, RBP-JK, Jagged1 and Msx2 protein decreased significantly in OPG intervention group (P < 0.05). Conclusion Osteoprotegerin is involved in the regulation of vascular calcification by the down regulating the expression of Notch1/RBP-JK pathway protein.