50 PEPTIDES A
100 80 60 40 20 0
60uM 40uM 20uM 10uM
■ Vitamin C ■ Meta TLR
B 40uM 60 40 20 0 CTRL MetaTlr TBHP(0.16mM)
Vitamin C 20 ug/ml
Figure 1: The senopreventive peptide exhibits strong antioxidant activity. (A) ABTS radical scavenge ratio of vitamin C and the senopreventive peptide. (B) Hydroxyl radical clearance rate of vitamin C and the senopreventive peptide. (C) ROS detection of HaCat induced by TBHP
SH-SY5Y and HaCaT co-cultures were challenged
with TNF-α (10 ng/mL) to induce neuroinflammatory responses. IL-1β, IL-6, and SP levels in supernatants were quantified using ELISA kits (Shenggong Biotech). TTBC (trans-4-tert-butylcyclohexanol, 1 µM) was applied as the positive control.
UVB-induced photoageing model HFF-1 were seeded in 12-well plates and incubated overnight with or without the Senopreventive Peptide. Cells were then exposed to UVB (25 mJ/ cm2
) and cultured for an additional 24 hours. Cells
were collected for the Senescence-associated β-galactosidase (SA-β-gal) activity test (Beyotime Biotechnology, Cat# C0602) or for RNA isolation and RT-qPCR for mRNA expression testing.
SPR test The binding affinity of the senopreventive peptide to TLR2 was assessed using SPR. TLR2 was immobilized on a CM5 sensor chip (GE Healthcare), and the senopreventive peptide (62.5–2000 nM) was injected at a flow rate of 30 μL/min. Binding affinity (Kd) was determined using Biacore Evaluation Software.
A
300 200 100 0
THP-1 cells *** *** ### B
100 80 60 40 20 0
LPS (10µg/mL)
60 40 20 0
C *** *** ###
900 600 300 0
-300
Normal TNF-α TTBC (1 µM) (50 µg/mL)
LPS (10µg/mL) TNF-α (10 ng/mL) MetaTlr
NF-κB activity test NF-κB reporter cell was modified based on Wilson’s method.10
Briefly, luciferase was replaced by dsRED,
which was driven by an NF-κB response element. So, the dsRED intensity represents NF-κB activity, while GFP driven by the UBC promoter served as a control. NF-κB reporter THP-1 cells were seeded in 96-
well plates and pre-treated with the senopreventive peptide (50 µg/mL) or dexamethasone for two hours. Cells were then stimulated with 100 ng/mL LPS for 24 hours, and the fluorescence intensity was measured using a fluorescence microscope (EVOS M5000) and a fluorescence microplate reader (Varioskan LUX).
Results The senopreventive peptide exhibits strong antioxidant activity Oxidative stress is a well-established contributor to skin ageing, leading to DNA damage, mitochondrial dysfunction, and the activation of inflammatory pathways. The antioxidant activity of the senopreventive peptide was first assessed using the ABTS assay. The senopreventive peptide
P815 mast cells ### *** ***
150 100 50 0
Normal SP SC MetaTlr
(50 µg/mL) (50 µg/mL) SP (1µM)
SY5Y and HaCaT co-cultured cells
** *** ### -50 Normal SP SC MetaTlr (50 µg/mL) (50 µg/mL) SP (1µM)
90 60 30 0
-30
Normal TNF-α TTBC MetaTlr (1 µM)
(50 µg/mL) TNF-α (10 ng/mL) ** ### **
900 600 300 0
** ### ***
30 20 10 0
* ###
showed similar activity to that observed with Vitamin C, and exhibited an 80% reduction in ABTS cation radical at a concentration of 60 µM. In addition, the hydroxyl radical scavenging
assay revealed that the Senopreventive Peptide exhibited activity comparable to vitamin C. Further evaluation in HaCaT cells demonstrated that both the senopreventive peptide and vitamin C significantly suppressed TBHP-induced ROS overproduction. These findings demonstrate the potent antioxidant capability of the senopreventive peptide, suggesting that it can effectively combat oxidative stress, a primary driver of ageing. The senopreventive peptide inhibits inflammatory cytokine secretion Inflammation plays a central role in the skin ageing process. In THP-1 macrophages, treatment with the senopreventive peptide significantly reduced the secretion of pro-inflammatory cytokines IL-6 and IL-1β following LPS stimulation, comparable to the effect of dexamethasone, a known anti- inflammatory agent. Similarly, in P815 mast cells, the senopreventive peptide inhibited the release of IL-6 and IL-1β and
C
*** Normal SP SC MetaTlr (50 µg/mL) (50 µg/mL) SP (1µM)
*** ### Normal SP *** SC MetaTlr (50 µg/mL) (50 µg/mL) TNF-α (10 ng/mL)
Figure 2: The senopreventive peptide exhibits anti-inflammatory effects caused by different stressors. (A) The senopreventive peptide inhibits LPS induced IL6/IL-1β expression in THP-1 cell. (B) The senopreventive peptide inhibits SP induced IL6/IL-1β expression and β-Hexosaminidase release in P815 Mast cell. (C) The senopreventive peptide inhibits TNF-α induced IL6/IL-1β and substance P expression in SH-SY5Y and HaCaT co-cultured cells
PERSONAL CARE MAGAZINE September 2026
www.personalcaremagazine.com
IL-6 level (ng/mL)
IL-1β level (ng/mL)
ABTS radical scavenge ratio %
Substance P (pg/mL)
The release rate of β-Hex (%)
Hydroxyl radical clearance rate (%)
IL-1β (ng/mL)
IL-1β (ng/mL)
IL-6 (ng/mL)
IL-6 (ng/mL)
Vitamin C Meta TIr
Normal LPS DEX-10ug/ml MetaTlr-20ug/ml
Normal LPS DEX-10ug/ml MetaTlr-20ug/ml
Page 1 |
Page 2 |
Page 3 |
Page 4 |
Page 5 |
Page 6 |
Page 7 |
Page 8 |
Page 9 |
Page 10 |
Page 11 |
Page 12 |
Page 13 |
Page 14 |
Page 15 |
Page 16 |
Page 17 |
Page 18 |
Page 19 |
Page 20 |
Page 21 |
Page 22 |
Page 23 |
Page 24 |
Page 25 |
Page 26 |
Page 27 |
Page 28 |
Page 29 |
Page 30 |
Page 31 |
Page 32 |
Page 33 |
Page 34 |
Page 35 |
Page 36 |
Page 37 |
Page 38 |
Page 39 |
Page 40 |
Page 41 |
Page 42 |
Page 43 |
Page 44 |
Page 45 |
Page 46 |
Page 47 |
Page 48 |
Page 49 |
Page 50 |
Page 51 |
Page 52 |
Page 53 |
Page 54 |
Page 55 |
Page 56 |
Page 57 |
Page 58 |
Page 59 |
Page 60 |
Page 61 |
Page 62 |
Page 63 |
Page 64 |
Page 65 |
Page 66 |
Page 67 |
Page 68 |
Page 69 |
Page 70 |
Page 71 |
Page 72 |
Page 73 |
Page 74 |
Page 75 |
Page 76 |
Page 77 |
Page 78 |
Page 79 |
Page 80 |
Page 81 |
Page 82 |
Page 83 |
Page 84 |
Page 85 |
Page 86 |
Page 87 |
Page 88 |
Page 89 |
Page 90 |
Page 91 |
Page 92
