Dual effects of ouabain on the regulation of proliferation and apoptosis in human umbilical vein endothelial cells: involvement of Na+-K+-ATPase α-subunits and NF-κB

作者：Ren, Yan-Ping^*; Zhang, Ming-Juan; Zhang, Ting; Huang, Ruo-Wen

来源：International Journal of Clinical and Experimental Medicine, 2014, 7(5): 1214-1222.

• Ouabain
• endothelium
• alpha subunit
• NF-kappa B

Purpose: To elucidate the effect of ouabain on the regulation of proliferation and apoptosis of HUVECs and involvement of different Na+-K+-ATPase alpha-subunits and NF-kappa B. Methods: HUVECs were isolated by collagenase perfusion, and MTT assays and cell cycle analysis were performed to study proliferation. NF-kappa B expression and function were examined by immunohistochemical staining and western blotting. Na+-K+-ATPase activity was determined by measuring released ouabain inhibitable inorganic phosphate (Pi). The expression of different alpha-subunits was investigated by real RT-PCR, western blotting and cell immunofluorescence. Results: 0.3 nM ouabain treatment for 0.5 h triggered the proliferation of HUVECs, peaking at 1-2 h. At 1.8 nM for 0.5 h, ouabain induced an increase of cell proliferation for a short time, and then triggered a decrease after 1 h. Cell cycle analysis show that 37% of HUVECs were in G2/M phase of the cell cycle following incubation with 1.8 nM ouabain, compared with 18% with 0.3 nM ouabain. NF-kappa B activity was assessed by western blot analysis of I kappa B expression, which was significantly reduced with 0.3 nM ouabain treatment; there was no different between 1.8 nM ouabain treatment and untreated cells. Na+-K+-ATPase activity in HUVECs was markedly reduced after treatment with 0.3 nM and 1.8 nM ouabain. Real RTPCR and western blotting indicated that Na+-K+-ATPase alpha(1)-subunit mRNA expression levels increased after 0.3 nM ouabain treatment and decreased after 1.8 nM ouabain treatment. However, alpha(2)- and alpha(3)-subunit mRNA decreased after 0.3 nM ouabain treatment and increased after 1.8 nM ouabain treatment. Conclusion: Ouabain at different concentrations caused dual effects on proliferation and apoptosis in HUVECs.

• 出版日期2014
• 单位西安交通大学