蒌叶果胶的提取及其抗皮肤老化与美白作用评价

Extraction of Piper Betel Leaf ( Piper betel L. ) Pectin and Evaluation of Its Anti Aging and Whitening Effects on Skin

摘要:

以纤维素酶解辅助超声-酸法提取蒌叶果胶，评估其抗皮肤老化和美白的潜力。以单因素试验和响应面法优化蒌叶果胶的提取工艺；以自由基清除试验和酶抑制实验考察其自由基清除能力和对酪氨酸酶、透明质酸酶抑制活性；通过UVB辐照HaCaT细胞构建衰老模型，以细胞活力检测和衰老β-半乳糖苷酶染色考察抗老化功效；通过斑马鱼幼鱼暴露试验评估其美白作用及生物安全性。结果表明，该工艺中酶解辅助的最佳条件为酶添加量3.0 wt.%，酶解时间119 min，温度60 ℃，此时果胶实际得率7.58±0.21%。蒌叶果胶在质量浓度8 mg·mL-1时对DPPH和OH-自由基的清除率分别为51.48%、62.40%；对透明质酸酶、酪氨酸酶有良好抑制作用，IC50分别为7.84 mg·mL-1、10.43 mg·mL-1；蒌叶果胶能减轻UVB造成的HaCaT细胞活力下降，降低衰老β-半乳糖苷酶阳性细胞比例；质量浓度150 μg·mL-1的蒌叶果胶使B16-F10细胞相对黑色素百分比和酪氨酸酶活性分别降至对照组的74.06%、75.69%；质量浓度3 mg·mL-1蒌叶果胶处理斑马鱼幼鱼，其体内相对黑色素百分比和酪氨酸酶活性分别降至对照组的60.52%和64.50%，且无毒性和刺激性。综上，酶解辅助超声-酸法提取的蒌叶果胶具有抗皮肤老化和美白作用，温和安全，有望开发为新型食品级天然护肤原料。

Abstract:

Piper betel leaf pectin was extracted via a cellulase-assisted ultrasound-acid method, and its potential for anti-skin photoaging and whitening was evaluated. The extraction process was optimized by combining single-factor experiments and response surface methodology. The free radical scavenging capacity and inhibitory activities against tyrosinase and hyaluronidase were investigated using free radical scavenging assays and enzyme inhibition experiments. A senescence model was established by UVB irradiation of HaCaT cells, and the anti-skin photoaging effects were evaluated via cell viability assay and senescence-associated β-galactosidase staining. The whitening effect and biosafety were assessed using zebrafish larva exposure tests. The results showed that the optimal conditions for cellulase assistance were as follows: enzyme addition amount 3.0 wt.%, enzymatic hydrolysis time 119 min, and temperature 60?℃, under which the actual yield of pectin reached 7.58±0.21%. At a concentration of 8 mg·mL?1, Piper betel leaf pectin exhibited scavenging rates of 51.48% and 62.40% for DPPH and ·OH radicals, respectively. It also exhibited significant inhibitory activities on hyaluronidase and tyrosinase, with IC?? values of 7.84 mg·mL?1 and 10.43 mg·mL?1, respectively. The pectin was able to mitigate the decrease in HaCaT cell viability induced by UVB and reduce the proportion of SA-β-gal-positive cells. When the mass concentration of Piper betel leaf pectin was 150 μg·mL?1, the relative melanin percentage and relative tyrosinase activity of B16-F10 cells decreased to 74.06% and 75.69% of the control group, respectively. In zebrafish larvae treated with 3 mg·mL?1 Piper betel leaf pectin, the in vivo relative percentage of melanin and tyrosinase activity were reduced to 60.52% and 64.50% of the control group, respectively, with no toxicity or irritation observed. In conclusion, the Piper betel leaf pectin extracted by the enzyme-assisted ultrasound-acid method exhibits anti-skin photoaging and whitening effects, is mild and safe, and holds promise for development as a novel food-grade natural skincare ingredient.