BMP7 overexpression lentiviral vector construction and its effect on calcification of mouse aortic smooth muscle cells

FU Shi-lin; YI Xue-jiao; PAN Wen-xu; YIN Chun; KANG Hua-li; QIAN De-hui

doi:10.11659/jjssx.10E023055

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BMP7 overexpression lentiviral vector construction and its effect on calcification of mouse aortic smooth muscle cells

Basic Research | 更新时间：2024-02-02

- BMP7 overexpression lentiviral vector construction and its effect on calcification of mouse aortic smooth muscle cells
- JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY Vol. 33, Issue 2, Pages: 95-99(2024)
- 作者机构：
  
  陆军军医大学第二附属医院心血管内科，重庆 400037
- 作者简介：
- 基金信息：
- DOI：10.11659/jjssx.10E023055
  CLC： R34;R329.2
- Published：25 February 2024，
  
  Received：13 October 2023，
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付仕林,易雪娇,潘文旭等.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(02):95-99.

FU Shi-lin,YI Xue-jiao,PAN Wen-xu,et al.BMP7 overexpression lentiviral vector construction and its effect on calcification of mouse aortic smooth muscle cells[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2024,33(02):95-99.
付仕林,易雪娇,潘文旭等.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(02):95-99. DOI： 10.11659/jjssx.10E023055.

FU Shi-lin,YI Xue-jiao,PAN Wen-xu,et al.BMP7 overexpression lentiviral vector construction and its effect on calcification of mouse aortic smooth muscle cells[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2024,33(02):95-99. DOI： 10.11659/jjssx.10E023055.

摘要

目的

构建骨形态发生蛋白7（BMP7）过表达慢病毒载体，分析BMP7过表达对小鼠主动脉内皮细胞Jagged1表达的影响及其对共培养血管平滑肌细胞（VSMCs）钙化的影响。

方法

根据目的基因信息Mouse-BMP7（NM_007557.3）和质粒信息pLVX-zsGreen-C1进行基因序列合成，构建BMP7过表达慢病毒。qPCR检测BMP7过表达慢病毒感染效率；Western blot检测其感染的小鼠主动脉内皮细胞Jagged1蛋白表达。将过表达BMP7慢病毒感染的内皮细胞与VSMCs共培养，茜素红染色观察共培养后VSMCs钙化情况。

结果

成功构建BMP7过表达慢病毒载体并转染至小鼠主动脉内皮细胞。qPCR检测结果提示，与正常对照组比较，BMP7过表达组的BMP7 mRNA相对表达量显著升高（

0.01），空载体对照组BMP7 mRNA无显著变化（

0.05）。Western blot结果显示，BMP7过表达组内皮细胞Jagged1蛋白表达水平较正常对照组显著降低（

0.01），而空载体对照组内皮细胞Jagged1蛋白水平较正常对照组无显著改变（

0.05）。茜素红染色结果提示，与感染过表达BMP7慢病毒的内皮细胞共培养后，VSMCs的钙化程度明显增加。

结论

成功构建小鼠BMP7过表达慢病毒载体，并发现过表达BMP7可以降低小鼠主动脉内皮细胞Jagged1表达，并促进共培养的VSMCs钙化。

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 （

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 （

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 （

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 （

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

Keywords

bone morphological protein 7lentiviral vector constructionJagged1vascular calcification

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