Effect of Antioxidants on the Fibroblast Replicative Lifespan In Vitro

Table 2

Effect of antioxidants on the fibroblast replicative lifespan in vitro.

Antioxidant	Cell line; concentration; effect on RLS	Other effects	Reference
1,9-Dimethyl-Methylene Blue, Toluidine Blue O, and Celestin Blue	IMR-90 fibroblasts; concentration not specified: no effect		[71]
18α-Glycyrrhetinic acid	HFL1 fibroblasts; 2 μg/ml; increase in RLS (58.5 to 64.3 PD)	Decrease in ROS level, Nrf2-mediated proteasome activation	[57]
2,2,6,6-Tetramethylpiperidine-1-oxyl (TEMPO)	IMR-90 fibroblasts; 25 μM: no effect; 100 μM: decrease of RLS (8–9PD)		[30]
3,3,5,5-Tetramethyl-l-pyrroline N-oxide	IMR-90 fibroblasts; 500 μM; slight increase in RLS (+1 PD)		[22]
3-Carbamoyl-2,2,5,5-tetramethylpyrrolidin-1-yloxy (3-CP)	IMR-90 fibroblasts; 25 μM; no effect; 100 μM: decrease of RLS (8–9PD)		[30]
4-Hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl (4-OH-TEMPO)	IMR-90 fibroblasts; 25 μM: no effect; 100 μM: decrease of RLS (8–9PD)		[30]
Ascorbic acid 2-phosphate	200 μM: HE49 cells, increase in RLS by 8.5 PD (20%) and 7.1 PD (2% O2); NYaKe cells, increase in RLS by 3.5 PD; WS3RGB cells, increase in RLS by 5.5 PD	Reduction of the rate of telomere shortening; decreased ROS level under 20% oxygen but not under 2% oxygen	[37]
Benzaldehyde	IMR-90 fibroblasts; 100 μM: no or toxic effect		[30]
Berberine	2BS and WI38 fibroblasts; 0.3125 μg/ml: no effect on RLS reported	Enhanced proliferation of cells during seven days of incubation. 3.5% SA-β-gal-positive cells seen in PD30 2BS cells, vs. 60% in late PDL cells; in berberine-treated 2BS cells (PD45), the SA-β-gal-positive cell rate reverted to 16%, as compared to the control group (56.5%).; Decreased level of p16; cyclin protein and cyclin-dependent kinases, such as cyclin D1 and CDK4 increased. Upregulated level of phosphorylated retinoblastoma protein (pRB)	[58]
Butylated hydroxytoluene	IMR-90 fibroblasts; 100 μM: no increase of RLS		[22]
Carnosine	HFF1 cells; 20 mM: RLS increased by 7.4 and 9.4 PD; 30 mM: RLS increased by 3.0, 4.3 and 6.3 PD; 50 mM: RLS decreased.; MRC-5 cells; 20 mM: RLS extension by 10 and 14 PD; 30 mM: RLS extension by 4 and 8 PD; 20 mM and 30 mM, introduced at 55 PD: RLS increase by 13.0 and 12.1 PD, respectively.; HFF-1 cells; 20 mM D-carnosine: no effect	50 mM carnosine: more flat, spread-out appearance; maintaining nonsenescent morphology	[66]
Chlorella vulgaris extract	Human diploid fibroblasts (HDFs); no effect of RLS reported	Young HDFs: reduced expression of SOD1, CAT, and CCS. Senescent cells: increased expression of the SOD2 and MAPK14 genes, downregulation of TP53 gene expression. Young and senescent HDFs: decreased expression of CDKN2A gene, increased expression of MAPK14 gene	[62]
Curcumin	H8F2p 25LM fibroblasts; 1: decrease of RLS		Sadowska-Bartosz et al., unpublished
Cyanidin	WI-38 cells; concentration not specified: increase of RLS (from 64 to 68 when added at 26 PD, from 62 to 65 when added at 42 PD, and from 62 to 63 when added at 58 PD)		[54]
Epigallocatechin gallate	Human diploid fibroblasts (HDF); 12.5 μM: effect on RLS not reported	Decrease of ROS level, increased mitochondrial potential, more intact mitochondrial DNA, elevated activities of antioxidative enzymes, more juvenile cell status	[53]
Ginsenoside Rg3(S)	HDFs maintained until different passages: PD 8-12 (young cells) and PD 34-36 (senescent cells). Senescent HDFs starved with serum-free DMEM overnight and then incubated in DMEM containing 10 or 30 μM Rg3(S) or (R) for 48 h: no effect on RLS reported	Decrease in percentages of SA-β-gal staining cells in replicatively old HDFs. Reversal of the replicative senescence of HDFs: restoration of the ATP level and NAD^+/NADH ratio in senescent HDFs. Recovering the cellular levels of ROS and the NAD^+/NADH ratio in young HDFs treated with rotenone. Downregulation of phosphatidylinositol 3-kinase/Akt through the inhibition of mTOR by cell cycle regulators like p53/p21 in senescent HDFs. Activation of sirtuin 3/PGC1α to stimulate mitochondrial biogenesis. Rg3(R) did not alter the corresponding signaling pathways	[61]
Kinetin	Primary cultures of human dermal fibroblasts; 40-200 μM: no effect on RLS	Cells retained some characteristic features of young cells	[65]
L-Sulforaphane	IMR-90 fibroblasts; 1 nM–10 μM: no effect		[71]
Malvidin	WI-38 cells; concentration not specified: increase of RLS (from 64 to 67 when added at 26 PD, from 62 to 66 when added at 42 PD, and from 62 to 63 when added at 58 PD)		[55]
Methylene Blue	IMR-90 fibroblasts; 100 nM: extension of RLS by >20 PD	Increase in ROS generation; concentration-dependent changes in the synthesis of complex IV and heme and iron uptake	[71]
N,N-Diphenyl-1,4-phenylenediamine	IMR-90 fibroblasts; 40 μM: no increase of RLS		[21]
N-Acetylcysteine	IMR-90 fibroblasts; 2 mM: no increase of RLS		[22]
N-Benzyl hydroxylamine	IMR-90 fibroblasts; 100 μM: RLS prolongation by at least 17–20 PD	Decrease in Rh123 accumulation, prevention of the age-dependent decline in the activity of aconitase	[30]
Nicotinamide	FB0603 fibroblasts; 0.5 and 5 mM: small increase in RLS (+2 PD)		[47]
Nicotinamide	IMR-90 fibroblasts; 3 mM: no significant effect at	Morphological rejuvenation of aged cells	[12]
Nicotinamide	NHFs; 5 mM from PD 30: significant extension of RLS	Decreased lipofuscin accumulation, decreased mitochondrial superoxide, decreased acidic β-galactosidase	[69]
Nitroso-tert-butane (tNB)	IMR-90 fibroblasts; 10 μM: no effect		[30]
N-Methyl hydroxylamine	IMR-90 fibroblasts; 100 μM: RLS prolongation by at least 17–20 PD	Decrease in Rh123 accumulation, prevention of the age-dependent decline in the activity of aconitase	[30]
N-t-Butyl hydroxylamine	IMR-90 fibroblasts; 10 μM: RLS prolongation by 5.8 PD; 100 μM: RLS prolongation by 19.7	Decrease in Rh123 accumulation, prevention of the age-dependent decline in the activity of aconitase	[30]
O-Benzyl hydroxylamine	IMR-90 fibroblasts; 100 μM: small decrease in RLS		[30]
O-Methyl hydroxylamine	IMR-90 fibroblasts; small decrease in RLS at 100 μM		[30]
O-t-Butyl hydroxylamine	IMR-90 fibroblasts; 100 μM: no effect		[30]
Piper betle (PB)	Primary HDFs derived from foreskins of three different male subjects aged between 9 and 12 years after circumcision; 0.4 mg/ml: no effect on RLS reported	Improvement of cell proliferation of young (143%), presenescent (127.3%) and senescent (157.3%) HDFs. Increased expressions of PRDX6, TP53, CDKN2A, PAK2, and MAPK14 in senescent HDFs. Modulation of the transcriptional profile changes in senescent HDFs. Increased expressions of SOD1, decreased expression of GPX1, PRDX6, TP53, CDKN2A, PAK2, and MAPK14 in PB-treated senescent HDFs	[59]
Quercetin caprylate, conc.	HFL1 fibroblasts; concentration not specified (≤5 μg/ml): a statistically significant increase in RLS, less than 5%	Maintenance of young morphology, delay in appearance of senescent phenotype, increase in growth rate; proteasome activation; morphological rejuvenation	[48]
Resveratrol	MRC-5 cells; 5 μM: small increase in RLS (+2 PD)	Reduced DNA breaks, attenuation of age-related increase in nuclear size, reduction in the level of acetylated forms of H3 and H4 histones and p53	[44]
Resveratrol	FB0603 cells; 0.2 and 1 μM: no effect on RLS		[47]
Resveratrol	H8F2p 25LM fibroblasts; 1 μM: decrease of RLS		Sadowska-Bartosz et al., unpublished
Resveratrol, oxyresveratrol, acetyl-resveratrol	Hs68 cells; 1-20 μM resveratrol, oxy-resveratrol, acetyl-resveratrol: decrease in RLS		[46]
Salicylic acid	IMR-90 fibroblasts; 200 μM: no increase of RLS		[22]
Thionine	IMR-90 fibroblasts; 1 μM: extension of RLS		[71]
Urolithin A	Human dermal fibroblasts; no effect of RLS reported	Significantly increased type I collagen expression and reduced matrix metalloproteinase 1 (MMP-1) expression; reduced intracellular ROS, partially due to activation of the Nrf2-mediated antioxidative response. 50 μM: changes in cell morphology and inhibition in cell proliferation, due to cell cycle arrest in G2/M phase. SA-β-gal and γH2AX unaltered	[56]
Vitamin C (ascorbic acid)	HEFs; extension of RLS by 2 PD (2 μM), 6 PD (20 μM), and 11 PD (200)	Decreased SA-β-gal staining, decreased ROS level, increase in aconitase activity, decrease in of p53, phospho-p53 at Ser15and p21 and apurinic/apyrimidinic site level, relieve of senescence-related G1 arrest	[35]
Vitamin E	WI-38 cells; 10 μg/ml and 100 μg/ml: increase in RLS (from 65 to 109 and 115, resp.)	Size and shape typical of replicatively young cells, decreased autofluorescence	[33]
Vitamin E	WI-38 cells; 10 μg/ml, 50 μg/ml and 100 μg/ml: no effect on RLS		[34]
α-Phenyl-t-butyl nitrone (PBN)	IMR-90 fibroblasts; 200 μM: increase in RLS from PD to PD		[22]
α-Phenyl-t-butyl nitrone (PBN)	IMR-90 fibroblasts; 200 μM: RLS prolongation by 2.4 PD; 800 μM: RLS prolongation by 14.8 PD		[30]
α-Tocopherol acetate	IMR-90 fibroblasts; 200 μM: no increase of RLS		[22]