caa a22 4500 606234462 CHVBK 20210128101237.0 cr unu---uuuuu 210128e20150701xx s 000 0 eng 10.1007/s11010-015-2418-y doi (NATIONALLICENCE)springer-10.1007/s11010-015-2418-y Roles of miRNA-24 in regulating endothelial nitric oxide synthase expression and vascular endothelial cell proliferation [Elektronische Daten] [Wenyu Zhang, Limei Yan, Yumei Li, Wei Chen, Nan Hu, Hui Wang, Hesheng Ou] This study is to investigate the effect of miRNA-24 on endothelial nitric oxide synthase (eNOS) expression and vascular endothelial cell proliferation. Constructed high expression miRNA-24 plasmids were introduced into human umbilical vein endothelial cells (HUVECs) by liposome. Cell numbers were counted by a Hemocytometer, and cell proliferation was detected by MTT assay. The expression levels of eNOS and specificity protein 1 (Sp1) at mRNA and protein levels were, respectively, examined by real-time PCR, immunohistochemistry, and Western blotting. Compared with the control group, endothelial cell proliferation in miRNA-24 group significantly decreased by 64.24% (6.53±0.11 vs 18.26±0.45, P<0.01). The expression of eNOS at mRNA and protein levels in miRNA-24 group decreased by 64.21% (0.34±0.01 vs 0.95±0.02, P<0.05) and 82.86% (0.072±0.06 vs 0.42±0.06, P<0.05), respectively. Meanwhile, the expression of Sp1 at mRNA and protein levels in miRNA-24 group decreased by 64.64% (0.35±0.01 vs 0.99±0.03, P<0.05) and 60.34% (0.23±0.05 vs 0.58±0.07, P<0.05), respectively. In anti-miRNA-24 group, endothelial cell proliferation decreased by 33.46% compared with the control group (12.15±0.21 vs 18.26±0.45, P<0.01), while it increased by 46.25% compared with the miRNA-24 group (12.15±0.21 vs 6.53±0.11, P<0.01). The expression of eNOS at mRNA and protein levels in anti-miRNA-24 group decreased by 44.21% (0.53±0.04 vs 0.95±0.02, P<0.05) and by 30.95%(0.29±0.05 vs 0.42±0.06, P<0.05) compared with the control group, while it increased by 35.84% (0.53±0.04 vs 0.34±0.01, P<0.05) and by 75.17% (0.29±0.05 vs 0.072±0.06, P<0.05) compared with miRNA-24 group. The expression of Sp1 at mRNA and protein levels in ant-miRNA-24 group decreased by 36.36% (0.63±0.04 vs 0.99±0.03, P<0.05) and by 22.41% (0.45±0.06 vs 0.58±0.07, P<0.05) compared with the control group, while it increased by 44.44% (0.63±0.04 vs 0.35±0.01, P<0.05) and by 48.88% (0.45±0.06 vs 0.23±0.05, P<0.05) compared with miRNA-24 group. HUVECs proliferation and eNOS expression are significantly inhibited by miRNA-24. Sp1, which is regulated by miRNA-24, might act as one of the important factors involved in eNOS gene expression. Springer Science+Business Media New York, 2015 miRNA-24 nationallicence Cell proliferation nationallicence Endothelial nitric oxide synthase nationallicence Sp1 nationallicence Zhang Wenyu College of Pharmacy, Guangxi Medical University, No. 22, Shuangyong Road, 530021, Nanning, People's Republic of China aut Yan Limei Biochemistry Center, University of South China, 421001, Hengyang, People's Republic of China aut Li Yumei College of Pharmacy, Guangxi Medical University, No. 22, Shuangyong Road, 530021, Nanning, People's Republic of China aut Chen Wei College of Pharmacy, Guangxi Medical University, No. 22, Shuangyong Road, 530021, Nanning, People's Republic of China aut Hu Nan College of Pharmacy, Guangxi Medical University, No. 22, Shuangyong Road, 530021, Nanning, People's Republic of China aut Wang Hui College of Pharmacy, Guangxi Medical University, No. 22, Shuangyong Road, 530021, Nanning, People's Republic of China aut Ou Hesheng College of Pharmacy, Guangxi Medical University, No. 22, Shuangyong Road, 530021, Nanning, People's Republic of China aut Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 405/1-2(2015-07-01), 281-289 0300-8177 405:1-2<281 2015 405 11010 https://doi.org/10.1007/s11010-015-2418-y text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s11010-015-2418-y text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Zhang Wenyu College of Pharmacy, Guangxi Medical University, No. 22, Shuangyong Road, 530021, Nanning, People's Republic of China aut NATIONALLICENCE 700 1- Yan Limei Biochemistry Center, University of South China, 421001, Hengyang, People's Republic of China aut NATIONALLICENCE 700 1- Li Yumei College of Pharmacy, Guangxi Medical University, No. 22, Shuangyong Road, 530021, Nanning, People's Republic of China aut NATIONALLICENCE 700 1- Chen Wei College of Pharmacy, Guangxi Medical University, No. 22, Shuangyong Road, 530021, Nanning, People's Republic of China aut NATIONALLICENCE 700 1- Hu Nan College of Pharmacy, Guangxi Medical University, No. 22, Shuangyong Road, 530021, Nanning, People's Republic of China aut NATIONALLICENCE 700 1- Wang Hui College of Pharmacy, Guangxi Medical University, No. 22, Shuangyong Road, 530021, Nanning, People's Republic of China aut NATIONALLICENCE 700 1- Ou Hesheng College of Pharmacy, Guangxi Medical University, No. 22, Shuangyong Road, 530021, Nanning, People's Republic of China aut NATIONALLICENCE 773 0- Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 405/1-2(2015-07-01), 281-289 0300-8177 405:1-2<281 2015 405 11010