| 摘要: | 多酚類具有許多生物活性如抗氧化，抑制MMPs 與彈力蛋白酶活性，若能抑制彈力蛋白酶或基質金屬蛋白酶之分泌或活性，則可延緩皺紋生成。本研究針對茜草科植物咖啡(Coffea arabica, CA)、檄樹(Morinda citrifolia, MC)和木蘭科植物玉蘭花(Michelia alba, MA)的葉萃取物及其水解物、咖啡成分綠原酸和咖啡酸等探討其抑制MMP 活性。上述三種植物以甲醇製備其萃取物，再以鹽酸水解製備水解物。三種植物之萃取物及其水解物先以膠原蛋白酶與彈力蛋白酶抑制實驗進行初篩。具活性者再以紫外線B 照射纖維母細胞誘導MMPs 產生之模式，探討對第I 型膠原蛋白原的生成及MMPs 之作用，進而再探討對MAP Kinases 磷酸化的影響。並針對具活性者進行安全性評估。
CA、MA 及MC 之甲醇萃取率分別為10.4%、16.9%、15.0%。於明膠分解初篩試驗結果顯示：咖啡萃取物(CAE，相當於原植物9.6mg/mL)與玉蘭花萃取物(MAE，相當於原植物5.9 mg/mL)及兩者的水解物(CAH, MAH 1000 μg/mL)抑制率皆大於90%。檄樹萃取物(MCE相當於原植物6.7 mg/mL)抑制率74.3%，水解物抑制率皆低於萃取物。而咖啡酸與綠原酸均無此抑制效果。
膠原蛋白酶活性抑制試驗結果顯示：CAE 具抑制效果，且具有濃度依存性，序列稀釋萃取物，相當於原植物濃度0.096~4.8 mg/mL 之抑制率84.6~100.3%。CAH 之抑制效果未優於CAE。MAE 具抑制效果，且具有濃度依存性，序列稀釋萃取物，相當於原植物濃度0.059~2.96 mg/mL 之抑制率92.7~100.1%。MAH 之抑制效果未優於MAE。彈力蛋白酶活性抑制試驗結果顯示：CAE 及CAH 皆無抑制效果。而MAE 與MAH，抑制效果亦不佳。對於UVB 所誘導MMP-1, -3, -9 之抑制表現試驗結果顯示，CAE能抑制MMP-1, -3, -9 之濃度分別為相當於原植物0.048, 0.96, 0.48mg/mL。MAE 能抑制MMP-1, -3 之濃度分別為相當於原植物0.029及0.059 mg/mL。咖啡酸能抑制MMP-1, -3 的表現之濃度分別為5 及50 μM。綠原酸能抑制MMP-1, -3 表現之濃度分別為10 及50 μM。CAE 能提高第I 型膠原蛋白原生成的濃度為相當於原植物0.096mg/mL。MAE 無提高生成的效果。CAE 可抑制JNK、ERK 及p38 磷酸化之濃度分別為相當於原植物0.048, 0.24 及0.048 mg/mL。MAE 可抑制JNK 及ERK 磷酸化之濃度分別為相當於原植物0.029 及 0.148 mg/mL。CAE 與MAE 於高濃度(相當於原植物4.8 及2.9 g/mL)無皮膚刺激性。兩者皆無眼球刺激性。此結果顯示咖啡及玉蘭花具有開發成為抗光老化化妝品之潛力。
Polyphenols are a group of compounds possessing a variety of
biological activities including anti-oxidant, inhibition of MMP-1 and
elastase. The inhibition of elastase and MMPs in the skin would slow
down the process of aging. In this study, Coffea arabica (CA), Morinda
citrifolia (MC) and Michelia alba (MA) extracts and their hydrolysates,
and active constituents of CA, chlororgenic acid and caffeic acid, had
been selected to screen for MMPs inhibition. The plants were extracted
with methanol and the extracts were hydrolyzed with different
concentration of hydrochloric acid. The chlororgenic acid, caffeic acid,
the various concentration of extracts and its hydrolysates were subject the
inhibition tests to MMP-1 and elastase. The fibroblasts were used for the
collagen synthesis promotion, MMPs inhibition and MAP Kinases
phosphorylation inhibition tests on plants extracts. The active ones were
process to safety assessment.
The methanolic extraction yield of CA, MA and MC were 10.4%,
16.9% and 15%, respectively. In gelatin digestion test, the inhibition
effects of CAE (equal to 9.6 mg dried leaves/mL), MAE (equal to 9 mg
dried leaves/mL) and their hydrolysates (1000 μg/mL CAH and MAH)
were more than 90%, in MCE (equal to 6.7 mg dried leaves/mL) was
74.3%. However, cholorogenic acid and caffeic acid show no effect on
gelatin digestion.
The results indicated that CAE would inhibit collagenase activity and present dose dependent effects. The inhibition results were in range of 84.6%~100.3% at the concentrations of 0.096~4.8 mg/mL (dried
leaves), respectively. The collagenase inhibition of CAE was superior to CAH. MAE would inhibit collagenase activity and present dose dependent effects. The inhibition results were in range of 92.7%~100.1%
at the concentrations of 0.059~2.96 mg/mL (dried leaves), respectively.
The effects of MAE on collagenase inhibition were better than MAH,
moreover, CAE, CAH, MAE and MAH had no significantly inhibition
results on elastase activity.
CAE would inhibit MMP-1, -3 and -9 at concentration of 0.048, 096
and 0.48 mg/mL (dried leaves) and MAE would inhibit MMP-1 and -3 at
0.029 and 0.059 mg/mL (dried leaves). Caffeic acid would suppress
MMP-1 and -3 expressions at 5 and 50 μM, moreover, that of chlorogenic
acid were 10 and 50 μM. In addition, CAE would elevate type I collagen
synthesis at 0.096 mg/mL (dried leaves), but not MAE.
In MAP Kinases inhibition, the results indicated that CAE would
inhibit the phosphorylation of JNK, ERK and p38 at the concentrations of
0.048, 0.24 and 0.048 mg/mL (dried leaves) and MAE would inhibit JNK
and ERK phosphorylation at 0.029 and 0.148 mg/mL. In safety
assessment, CAE and MAE did not present toxicity at higher
concentrations (equal to 4.8 and 2.9 mg dried leaves/mL) on the skin and
eye irritation test.
The results show that Coffea arabica and Michelia alba could
become the promising anti-photoaging material of cosmetics through this
investigation. |
