ANTI-AGING COMPOUNDS FROM JEJU ISLAND 27
Table II (continued)
DPPH radical scavenging activity (%)
Plant: Botanical name 400 200 100 50 10 SCso *
(part usedt µg/ml µg/ml µg/ml µg/ml µg/ml µg/ml (µg/ml)
Potamogeton crispus L. (WP) 87.9 74 39.4 65
Pyrrosia hastasts (LT) 77.2 75.3 42.3 9.3 61.6
Pyrrosia hastata (RT) 81.5 83.9 79.6 19.9 30.1
Sasa quelpaertensis (LT) 50 14.9 400
Scirpus tabernaemontani (LT) 85.2 78.5 16.7 2.2 31.5
Solanum nigrum L. (WP) 400
Solanum tuberosum L. (RH) 30.1 3.7 400
Thymus quinquecostatus (WP) 79.8 52.8 26.7 94.6
Torreya nucifera (LF) 90.4 89.2 31.9 6.5 22.7
T orreya nucifera (ST) 72.6 46.4 28.9 112.8
Typha orientalis (SD) 89.6 88.9 34.1 6.7 21.6
Zingiber mioga (ST) 30.2 400
BHT 82.5 64.1 33.1 6.2 25.4
a Part used: LF (leaf), ST (stem), LT (leaves and twigs), RT (root), WP (whole plant), SD (seed), FL (flower),
RH (rhizome).
*SC
50
Concentration (µg/ml) at which the inhibition %of DPPH radical scavenging activity is 50%.
BHT: di-t-butyl hydroxyl toluene.
ASSAY ON MMP-1 EXPRESSION BY RT-PCR
It was mentioned above that the major component of ECM in the dermis is collagen
fiber, and ECM degradation is associated with the generation of wrinkles. The enzymes
related to collagenase mRNA, and which degrade collagen fiber, are known as MMPs
(matrix metalloproteinases) (4). They can be divided into four main classes: collagenase
(MMP-1,8,13,18), gelatinase (MMP-2, 9), stomelysin (MMP-3,10), and membrane-type
MMPs (MMP-14,15,16,17), (7,8). Since Gross and Lapiere reported the ability of
MMP-1 to degrade the collagen triple helix 1962 (9), it has been used in the evaluation
of collagen degradation activity by estimating the amount of MMP-1 mRNA. In this
study, in order to screen anti-aging candidates that would inhibit the degradation of
collagen fiber, we investigated the reduction of MMP-1 expression for seven extracts
with superior elastase inhibition activity using the RT-PCR method. EGCG was used
as a positive control because its activities were well-known for the inhibitory effect on
collagenase and stromelysin mRNA expression induced by L-1� (25) and for the pro-
tective effect against skin damage due to UVB (26).
The MMP-1 expression assay in human fibroblasts was carried out with a gel image
analyzer (Bio-Rad, Gel DOC 2000). The relative amounts of MMP-1 expression extracts
were compared at a concentration of 100 µg/ml, and the results from seven plants are
shown in Figure 4. The results showed that MMP-1 expression rates of Typha orientalis
(seed), Pyrrosia hastata (root), and Capsicum annum (whole plant) were a little higher or
similar to that of EGCG, while four extracts, Persicaria hydropiper (whole plant), Fili-
pendula glaberrima (root), Nymphaea tetragona (rhizome), and Camellia japonica (leaf),
showed up to 100% inhibition of MMP-i expression.
Table II (continued)
DPPH radical scavenging activity (%)
Plant: Botanical name 400 200 100 50 10 SCso *
(part usedt µg/ml µg/ml µg/ml µg/ml µg/ml µg/ml (µg/ml)
Potamogeton crispus L. (WP) 87.9 74 39.4 65
Pyrrosia hastasts (LT) 77.2 75.3 42.3 9.3 61.6
Pyrrosia hastata (RT) 81.5 83.9 79.6 19.9 30.1
Sasa quelpaertensis (LT) 50 14.9 400
Scirpus tabernaemontani (LT) 85.2 78.5 16.7 2.2 31.5
Solanum nigrum L. (WP) 400
Solanum tuberosum L. (RH) 30.1 3.7 400
Thymus quinquecostatus (WP) 79.8 52.8 26.7 94.6
Torreya nucifera (LF) 90.4 89.2 31.9 6.5 22.7
T orreya nucifera (ST) 72.6 46.4 28.9 112.8
Typha orientalis (SD) 89.6 88.9 34.1 6.7 21.6
Zingiber mioga (ST) 30.2 400
BHT 82.5 64.1 33.1 6.2 25.4
a Part used: LF (leaf), ST (stem), LT (leaves and twigs), RT (root), WP (whole plant), SD (seed), FL (flower),
RH (rhizome).
*SC
50
Concentration (µg/ml) at which the inhibition %of DPPH radical scavenging activity is 50%.
BHT: di-t-butyl hydroxyl toluene.
ASSAY ON MMP-1 EXPRESSION BY RT-PCR
It was mentioned above that the major component of ECM in the dermis is collagen
fiber, and ECM degradation is associated with the generation of wrinkles. The enzymes
related to collagenase mRNA, and which degrade collagen fiber, are known as MMPs
(matrix metalloproteinases) (4). They can be divided into four main classes: collagenase
(MMP-1,8,13,18), gelatinase (MMP-2, 9), stomelysin (MMP-3,10), and membrane-type
MMPs (MMP-14,15,16,17), (7,8). Since Gross and Lapiere reported the ability of
MMP-1 to degrade the collagen triple helix 1962 (9), it has been used in the evaluation
of collagen degradation activity by estimating the amount of MMP-1 mRNA. In this
study, in order to screen anti-aging candidates that would inhibit the degradation of
collagen fiber, we investigated the reduction of MMP-1 expression for seven extracts
with superior elastase inhibition activity using the RT-PCR method. EGCG was used
as a positive control because its activities were well-known for the inhibitory effect on
collagenase and stromelysin mRNA expression induced by L-1� (25) and for the pro-
tective effect against skin damage due to UVB (26).
The MMP-1 expression assay in human fibroblasts was carried out with a gel image
analyzer (Bio-Rad, Gel DOC 2000). The relative amounts of MMP-1 expression extracts
were compared at a concentration of 100 µg/ml, and the results from seven plants are
shown in Figure 4. The results showed that MMP-1 expression rates of Typha orientalis
(seed), Pyrrosia hastata (root), and Capsicum annum (whole plant) were a little higher or
similar to that of EGCG, while four extracts, Persicaria hydropiper (whole plant), Fili-
pendula glaberrima (root), Nymphaea tetragona (rhizome), and Camellia japonica (leaf),
showed up to 100% inhibition of MMP-i expression.
