Biochemical and Biophysical Research Communications
Anti-apoptotic and anti-senescence effects of Klotho on vascular endothelial cells
Section snippets
Methods
Materials. A mouse membrane-form Klotho cDNA was kindly donated by Professor Yoichi Nabeshima (Department of Pathology and Tumor Biology, Kyoto University Graduate School of Medicine). The PCAGGS plasmid was kindly donated by Professor Jun-ichi Miyazaki (Division of Stem Cell Regulation, Osaka University Graduate School of Medicine) [22]. Anti-Klotho rat monoclonal antibody (KM2076) was kindly donated by Kyowa Hakko Kogyo (Tokyo, Japan) [23]. All other materials were commercial products of the
Transfection efficiency and Klotho expression in COS-1 cells
To investigate the efficiency of COS-1 cell transfection, we examined the expression of Klotho protein in cell lysates from COS-1 cells. As reported recently [26], Klotho was expressed abundantly in COS-1 cells without transfection, as was demonstrated in IMCD cells (data not shown). In Klotho-transfected COS-1 cells, the expression of Klotho protein was dramatically increased (data not shown).
Klotho overexpression reduced apoptosis in COS-1 cells
It was reported that apoptotic cells were detected in the hippocampus in KL−/− mice at the age of 7
Discussion
This is the first report that purified Klotho recombinant protein has anti-apoptotic and anti-senescent effects in endothelial cells. Kuro-o et al.[1] reported that Klotho mutant mice manifest multiple age-related disorders that are also observed in humans, including atherosclerosis. It is well known that apoptosis and senescence in vascular endothelial cells initiate atherosclerosis. It has also been reported that apoptotic cells can be detected in the hippocampus in Klotho mutant mice at the
Acknowledgments
We thank Ms. Seiko Kaji for her excellent technical assistance. This work was supported by the Osaka Medical Research Foundation for Incurable Diseases (2000, 2005), NOVARTIS Foundation for Gerontological Research (2001), and Grants-in-Aid for scientific research from the Ministry of Education, Science, Sports, Culture and Technology of Japan (2003, 2004).
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