骨髓间充质干细胞对钙化血管平滑肌细胞的调控作用  

Effects of bone marrow mesenchymal stem cells on the mineralization of calcified vascular smooth muscle cells

作　　者：朱蒙恩[1] 方欣[1] 管思明[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院老年病科,武汉430022

出　　处：《中华老年医学杂志》2016年第11期1225-1228,共4页Chinese Journal of Geriatrics

摘　　要：目的研究骨髓间充质干细胞（MSC）对钙化血管平滑肌细胞（CSMC）的调控作用。方法以地塞米松、p甘油磷酸钠、维生素C作为钙化诱导剂（OS）诱导血管平滑肌细胞（SMC）钙化从而建立CSMC模型，采用VonKossa染色法评估钙化情况。随后将MSC与CSMC以1：1的比例分别培养于12孔Transwell板上室和下室，建立MSC干预CSMC模型。继而将SMC分为4组：正常组：正常SMC；钙化诱导剂组：SMC＋OS；钙化细胞组：CSMC；MSC干预组：CSMC＋MSC。其中，钙化诱导剂组应用OS培养基培养，其余各组均使用普通培养基培养。干预14d后收集各组SMC，检测碱性磷酸酶（AKP）活性评估SMC钙化情况，Westernblot技术检测SMC中Wnt5a及β-catenin蛋白表达。结果与正常组比较，钙化诱导后CSMC矿化结节明显增加，且钙化诱导剂组及钙化细胞组AKP活性、Wnt5a及β-catenin蛋白表达水平均明显升高。钙化细胞组与钙化诱导剂组比较，AKP活性、Wnt5a及β-catenin蛋白表达水平有所降低。不论与钙化诱导剂组还是钙化细胞组比较，MSC干预组上述检测指标均明显降低。结论MSC与CSMC间接接触可降低CSMC钙化程度，其可能通过旁分泌途径阻断Wnt5a／β-catenin通路从而减轻钙化。Objective To study the effects of bone marrow mesenchymal stem cells（MSC）on the mineralization of calcified vascular smooth muscle cells （CSMC） . Methods Osteogenie differentiation of vascular smooth muscle cells （SMC）was induced by osteogenic medium for 14 days. Then von Kossa staining was performed to observe the mineral deposits in SMC. The transwell system was used to establish the indirect co-culture environment. And hence,CSMC was indirectly co- cultured without or with MSC at a ratio of 1 ： 1 in the down plates or up plates. The cells were divided into 4 groups. Normal group included the normal SMC cultured with the normal medium. Calcification inducers group included the normal SMC cultured with the osteogenic medium. Calcified cells group included CSMC cultured with the normal medium. MSC intervention group included MSC and CSMC cultured with the normal medium. 14 days later, SMC from 4 groups was harvested. Alkaline phosphatase（AKP）aetivity was detected to evaluate the severity of calcification. Moreover,the protein levels of Wnt5a and β-catenin were measured by Western blot analysis. Results Compared with normal group,the calcification inducers group showed that AKP activity,WntSa and β-catenin protein levels were notably increased. Additionally, AKP activity and the protein levels of WntSa, β-catenin were decreased in calcified cells group as compared with calcification inducers group. Furthermore, AKP activity and the protein levels of WntSa, β--catenin were significantly reduced （P〈0.01）in MSC intervention group versus calcified ceils group Conclusions When MSC is indirectly co-cultured with CSMC,MSC can relieve the vascular calcification, which might be mediated by blocking the Wnt5a/β- catenin signaling through immunomodulatory and paracrine.

关 键 词：间质干细胞 肌 平滑 血管 钙化 

分 类 号：R54[医药卫生—心血管疾病]