Knockdown of paraoxonase 1 expression influences the ageing of human dermal microvascular endothelial cells.
Skin is one of the most commonly studied tissues for microcirculation research owing to its close correlation of cutaneous vascular function, ageing and age-related cardiovascular events. To elucidate proteins that determine this correlation between endothelial cell function and ageing in the vascular environment of the skin, we performed a proteomic analysis of plasma samples from six donors in their 20s (young) and six donors in their 60s (old). Among identified proteins, paraoxonase 1 (PON1) was selected in this study. To elucidate the role of PON1 on skin ageing and determine how it controls cellular senescence, the characteristics of PON1 in human dermal microvascular endothelial cells (HDMECs) were determined. When the expression of endogenous PON1 was knocked-down by small interfering RNA (siRNA) targeting PON1, HDMECs showed characteristic features of cellular senescence such as increases in senescence-associated β-galactosidase stained cells and enlarged and flattened cell morphology. At 48 h post-transfection, the protein expression of p16 in PON1 siRNA-treated HDMECs was higher than that in scrambled siRNA-treated HDMECs. In addition, the expressions of moesin and rho GTP dissociation inhibitor, additional age-related candidate biomarkers, were decreased by PON1 knock-down in HDMECs. In conclusion, these results suggest that PON1 functions as an ageing-related protein and plays an important role in the cellular senescence of HDMECs.
Department of Dermatology, Cutaneous Biology Research Institute, Yonsei University College of Medicine, Seoul, Korea.
Yun Sun
YS
Lee
Chang Ook
CO
Park
Ji Yeon
JY
Noh
Shan
S
Jin
Na Ra
NR
Lee
Seongmin
S
Noh
Ju Hee
JH
Lee
Kwang Hoon
KH
Lee
D000328:Adult; D000368:Aged; D000375:Aging / Q000235:genetics / Q000378:metabolism; D043303:Aryldialkylphosphatase / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D015415:Biomarkers / Q000378:metabolism; D002478:Cells, Cultured; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000378:metabolism; D042783:Endothelial Cells / Q000378:metabolism* / Q000473:pathology; D055785:Gene Knockdown Techniques; D006801:Humans; D008840:Microfilament Proteins / Q000378:metabolism; D055806:Microvessels / Q000378:metabolism; D008875:Middle Aged; D034741:RNA, Small Interfering / Q000494:pharmacology; D012867:Skin / Q000098:blood supply / Q000378:metabolism*; D012879:Skin Physiological Phenomena / Q000187:drug effects / Q000235:genetics; D014162:Transfection; D055815:Young Adult; D001616:beta-Galactosidase / Q000378:metabolism; D062947:rho-Specific Guanine Nucleotide Dissociation Inhibitors / Q000378:metabolism
D015415:Biomarkers; D019941:Cyclin-Dependent Kinase Inhibitor p16; D008840:Microfilament Proteins; D034741:RNA, Small Interfering; D062947:rho-Specific Guanine Nucleotide Dissociation Inhibitors; C071058:moesin; D043303:Aryldialkylphosphatase; C468406:PON1 protein, human; D001616:beta-Galactosidase