目的研究结缔组织生长因子(CTGF)对小鼠血管平滑肌细胞(VSMCs)钙化影响。方法从小鼠胸主动脉得到VSMCs,以免疫细胞化学方法鉴定。CTGF处理VSMCs后,用0.1%茜素红矿化结节染色,细胞钙沉积含量测细胞钙盐沉积,实时定量PCR和Western印迹法检测成骨细胞标志物碱性磷酸酶(ALP)、骨钙素、骨保护素(OPG)、核结合因子d1(Cbfα-1/Runx2)的表达。结果CTGF可促进VSMCs钙盐沉积,增加ALP、骨钙素和OPG的mRNA表达,分别为对照组的(7.82±0.46)、(3.32±0.40)、(5.22±0.56)倍(均P〈0.05),同时CTGF也增加Cbfα-1/Runx-2的表达,呈浓度依赖性。结论CTGF可能通过诱导VSMCs转分化为成骨细胞而促进血管钙化。
Objective To investigate the effect of connective tissue growth factor (CTGF) on the calcification of mouse vascular smooth muscle cells (VSMCs). Methods VSMCs used in the present study were obtained from the thoracic aortas of mouse and were identified by immunohistochmistry. After the cells were treated with CTGF, mineralized matrix staining was performed with O. 1% Alizarin red, the cellular mineral deposition was measured by testing the concentration of calcium, and the expressions of bone markers such as alkaline phosphatase (ALP) , osteocalcin (OC) , osteoprotegerin (OPG) , and core-binding factor α1 (Cbfα-1/Runx2) mRNA and protein were detected by real-time PCR and Western blot. Results After treatment with CTGF for 14 days, the calcium level in VSMCs was increased, and the mRNA expressions of ALP, OC, and OPG were increased as well, being ( 7.82 ± 0. 46 ) , ( 3.32 ±0. 40 ) , and ( 5.22± 0.56 ) folds ( all P 〈0. 05 ). Meanwhile, CTFG stimulated the expression of Cbfα-1/Runx2 in a dose-dependent manner. Conclusion CTGF may stimulate the osteoblast phenotypic switch of VSMCs and accelerate vascular calcification.