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目的研究人第10号染色体缺失的磷酸酶及张力蛋白同源基因(phosphatase and tensin homolog deleted on chromosome 10,PTEN)在小鼠动脉血管钙化形成过程中的作用。方法 1构建血管特异性PTEN敲除小鼠(PTEN△/△),对照组小鼠为PTENf/f;2免疫组化检测Apo E全敲除小鼠钙化血管中PTEN的表达水平;3体外通过钙化培养基诱导血管钙化;4Alizarin Red染色和Von Kossa染色评估PTEN敲除后小鼠血管钙化程度;5实时荧光定量PCR检测血管钙化标志物Runx2、骨钙蛋白和BMP2的表达水平。结果 1与普通饮食组相比,高脂饮食诱导的Apo E基因敲除小鼠血管钙化明显,而钙化后的血管中PTEN的表达水平显著下降(P<0.01);2经体外诱导后,PTEN△/△小鼠血管明显钙化,而PTENf/f小鼠血管几乎无钙化;3与PTENf/f组相比,PTEN△/△小鼠血管钙化标志物Runx2、骨钙蛋白和BMP2的表达均明显升高(P<0.05)。结论血管特异性PTEN基因敲除促进小鼠血管钙化的形成。
Objective To study the role of phosphatase and tensin homolog deleted on chromosome 10 (PTEN) in the process of arterial calcification in mice. Methods 1 PTEN knockout mice (PTEN △ / △) were constructed, and PTENf / f mice were used as control group. 2 Expression of PTEN was detected by immunohistochemistry in calcified blood vessels of ApoE knockout mice. Calcification medium induced vascular calcification; 4Alizarin Red staining and Von Kossa staining were used to evaluate the degree of vascular calcification in PTEN knockout mice; 5 Real-time fluorescent quantitative PCR was used to detect the expressions of Runx2, osteocalcin and BMP2. Results 1Compared with the normal diet group, the vascular calcification of ApoE knockout mice induced by high-fat diet was significant, while the expression of PTEN in calcified blood vessels was significantly decreased (P <0.01) .2 After induction in vitro, PTEN Compared with PTENf / f group, the expressions of Runx2, osteocalcin and BMP2 in PTEN △ / △ mice were significantly higher than those in PTENf / f group Increased (P <0.05). Conclusion Vascular specific PTEN gene knockdown promotes the formation of vascular calcification in mice.