摘要
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 effect of phosphatase and tensin homolog deleted on chromosome 10 （PTEN） in mice vascular calcification. Methods Conditional deletion of PTEN in mice （PTEN △/△ ） was generated by crossing PTEN exon 5 floxed mice （PTEN f/f） with the SM22 α-Cre ＋/- transgenic mice. PTEN expression was detected by immunohistochemical staining in apolipoprotein E （ApoE -/- ） mice fed by high fat diet （HFD） or chow diet （CD）. Aortic rings excised from 4-week-old PTEN △/△ mice were cultured in calcified media. After 3 weeks ex vivo culture, arterial calcification was determined by Alizarin Red staining and Von Kossa staining in PTEN△/△ and control PTENf/f mice. The expression of runt related transcription factor 2 （ Runx2）, osteocalcin and bone morphogenetic protein 2 （BMP2） were detected by real-time PCR in both mice groups. Results Compared to CD fed group, HFD induced vascular calcification in ApoE -/- mice while PTEN expression was significantly decreased in the calcified arteries （P 〈 0. 01 ）. Immunostaining demonstrated obvious vascular calcification in PTEN△/△ mice after ex vivo stimulated culture, which was not evident ：in PTENf/f mice under the same condition. Furthermore, elevated mRNA expression of Runx2, osteocalc, in and BMP2 was detected in PTEN△/△ mice compared with PTENf/f mice by PCR analysis （P 〈 0.05）. Conclusion Muscle-specific deletion of PTEN promotes vascular calcification in mice.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2016年第24期2587-2591,共5页
Journal of Third Military Medical University
基金
国家自然科学基金青年科学基金(81400612)~~
关键词
动脉血管钙化
PTEN
RUNX2
骨钙蛋白
BMP2
vascular calcification
phosphatase and tensin homolog deleted on chromosome 10
Runx2
osteocalcin
bone morphogenetic protein 2
