Abstract
Free fatty acids (FFA)-induced proliferation and apoptosis was studied in human umbilical vein endothelial cells (HUVECs). A recombinant adenovirus containing a RNAi cassette targeting the GSK-3β gene was produced and its silencing effect on GSK-3β gene was detected by Western blot analysis and immunohistochemistry assay in HUVECs. The effect of the RNAi on the protein level of β-catenin was explored by transfecting the RNAi adenovirus to inhibit the expression of GSK-3β protein. The subsequent effect on the Wnt/GSK-3β/β-catenin signal pathway and on proliferation and apoptosis of HUVECs cultured with FFAs, was analyzed by BrdU assay, Annexin V-FITC/PI Apoptosis Detection Kit, and 4′,6-diamidino-2- phenylindole(DAPI) to explore the possible connection between the signaling pathway and FFA-induced proliferation and apoptosis. The Western blot results showed that the expression of GSK-3β protein in HUVECs could be inhibited efficiently by the RNAi adenovirus, and that the protein level of β-catenin was increased by RNAi adenovirus transfection. The results of the BrdU assay suggested that knockdown of GSK-3β with the RNAi adenovirus may stimulate the proliferation of HUVECs. Apoptosis was observed in HUVECs exposed to FFAs (0.75 mmol/L) for 72 h, and this effect could be partly reversed when interfering with the RNAi adenovirus. It may be concluded that the RNAi adenovirus specific to GSK-3β may partly protect HUVECs from apoptosis induced by FFAs, probably through the up-regulation of the Wnt/β-catenin signal pathway.
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Acknowledgments
This study was supported by Grant C071002 and 2009Y0011 for natural science foundation from Fujian province of China and grant from the Ministry of Health of the People’s Republic of China (WKJ 2005-2-021). We thank the members of Department of Molecular Medicine Research Center lab of Fujian Medical University, specifically Drs Lin.
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The authors Penli Zhu, Gang Chen, Tingting You, Jin Yao, and Qiqin Jiang contributed equally to this work.
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Zhu, P., Chen, G., You, T. et al. High FFA-induced proliferation and apoptosis in human umbilical vein endothelial cell partly through Wnt/β-catenin signal pathway. Mol Cell Biochem 338, 123–131 (2010). https://doi.org/10.1007/s11010-009-0345-5
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DOI: https://doi.org/10.1007/s11010-009-0345-5