摘要
目的 观察促红细胞生成素(EPO)对钙化血管平滑肌细胞(VSMCs)中α-Klotho蛋白表达的影响,研究α-Klotho在EPO减轻血管钙化中的作用。方法 采用高磷刺激大鼠VSMCs钙化,实时聚合酶链反应检测钙化相关基因的表达变化,von Kossa染色和钙含量检测钙化情况,应用siRNA技术敲低VSMCs中α-Klotho的表达,Western blotting检测VSMCs中α-Klotho的蛋白表达。结果 与高磷刺激的钙化模型组相比,EPO下调了钙化VSMCs中骨钙素和Cbfα-1的mRNA水平(P <0.01),并减少了钙盐沉积和钙含量(P <0.01)。Western blotting结果发现,EPO可使钙化VSMCs中α-Klotho蛋白表达水平上调1.19倍(P <0.05)。敲低VSMCs中α-Klotho后,EPO抑制骨钙素和Cbfα-1 mRNA表达的作用下调。而且,α-Klotho敲低使EPO抑制钙盐沉积和钙含量的作用被阻断。结论 α-Klotho参与了EPO减轻VSMCs钙化的作用,该实验结果为防治血管钙化提示了可能的干预靶点。
Objective To observe the effect of erythropoietin(EPO) on α-Klotho protein expression in calcified vascular smooth muscle cells(VSMCs), and to analyze the role of α-Klotho in reducing vascular calcification. Methods High phosphorus was used to stimulate VSMCs calcification. The mRNA levels of calcification-related markers were detected by real-time PCR, and the calcium deposition was observed by von Kossa staining. α-Klotho was knocked down by siRNA. The α-Klotho protein expression was detected by Western blotting. Results EPO decreased the mRNA levels of osteocalcin and Cbfα-1 in calcified VSMCs(P <0.01). Compared with the high phosphorus group, calcium deposition and calcium content(P < 0.01) were down-regulated by EPO. Western blotting showed that EPO could increase the protein level of α-Klotho in calcified VSMCs by 1.19 fold(P < 0.05). Knockdown of α-Klotho in VSMCs decreased the inhibitory effect of EPO on osteocalcin and Cbfα-1 mRNA expression. After α-Klotho was knocked down, the effects of EPO inhibiting calcium deposition and calcium content were blocked. Conclusion α-Klotho is involved in the role of EPO in reducing VSMCs calcification, thus providing a new strategy for the prevention and treatment of vascular calcification.
作者
常晋瑞
魏明
曹健
南瑛
王倩
孙娜
Chang Jinrui;Wei Ming;Cao Jian;Nan Ying;Wang Qian;Sun Na(Institute of Basic Medicine Science,Xi'an Medical University,Xi'an 710021,China)
出处
《兰州大学学报（医学版）》
2023年第1期6-11,共6页
Journal of Lanzhou University（Medical Sciences）
基金
陕西省教育厅科研计划项目(20JK0883,20JS135)。
