局解手术学杂志2024，Vol.33Issue(2)：95-99,5.DOI:10.11659/jjssx.10E023055
BMP7过表达慢病毒载体构建及其对小鼠主动脉平滑肌细胞钙化的影响
BMP7 overexpression lentiviral vector construction and its effect on calcification of mouse aortic smooth muscle cells
摘要
Abstract
Objective To construct a lentiviral vector for overexpression of bone morphogenetic protein 7(BMP7)in mice,and the effect of BMP7 overexpression on the expression of Jagged1 in mouse aortic endothelial cells and the calcification of the co-cultured vascular smooth muscle cells(VSMCs)were analyzed.Methods According to the target gene information Mouse-BMP7(NM_007557.3)and plasmid information pLVX-zsGreen-C1,gene sequence synthesis was carried out to construct BMP7 overexpression lentivirus.The efficiency of BMP7 overexpression lentivirus infection was detected by qPCR;the expression of Jagged1 protein in aortic endothelial cells from infected mice was detected by Western blot.The endothelial cells with lentivirus overexpressing BMP7 were co-cultured with VSMCs,and the calcification of VSMCs was observed by alizarin red staining.Results BMP7 overexpression lentiviral vector was successfully constructed and transfected into aortic endothelial cells.qPCR test results showed that the expression level of BMP7 mRNA was significantly increased in the BMP7 overexpression group than that in the normal control group(P<0.01),while there was no significant difference in the expression of BMP7 mRNA between the empty vector control group and the normal control group(P>0.05).Western blot results showed that the expression level of Jagged1 protein in endothelial cells of mouse in the BMP7 overexpression group was significantly lower than that in the normal control group(P<0.01),while there was no significant difference in the expression level of Jagged1 protein in endothelial cells between the empty vector control group and the normal control group(P>0.05).The results of alizarin red staining showed that the calcification of VSMCs was significantly increased after co-cultured with endothelial cells infected with BMP7 lentivirus.Conclusion Mouse BMP7 overexpression lentiviral vector was successfully constructed,and overexpression of BMP7 can reduce the expression of Jagged1 in mouse aortic endothelial cells and promote the calcification of co-cultured VSMCs.关键词
骨形态发生蛋白7/慢病毒载体构建/Jagged1/血管钙化Key words
bone morphological protein 7/lentiviral vector construction/Jagged1/vascular calcification分类
医药卫生引用本文复制引用
付仕林,易雪娇,潘文旭,尹纯,康华利,钱德慧..BMP7过表达慢病毒载体构建及其对小鼠主动脉平滑肌细胞钙化的影响[J].局解手术学杂志,2024,33(2):95-99,5.基金项目
重庆市自然科学基金面上项目(CSTB2023NSCQ-MSX0686) （CSTB2023NSCQ-MSX0686）
陆军军医大学医院青年博士人才孵化计划项目(2022YQB029) （2022YQB029）
