In vitro anti-aging activities of ginkgo biloba leaf extract and its chemical constituents

Abstract To determine in vitro anti-aging activity and identify the active chemical constituents in Ginkgo leaf extracts. The antioxidant properties of Ginkgo biloba leaves extracts were evaluated using 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) scavenging and 2,2’-azino-bis (3-ethylbenzothiazoline)-6 sulfonic acid radical (ABTS) scavenging assays. The inhibitory effects of extracts and chemical constituents on reactive oxygen species, matrix metalloproteinase-1 (MMP-1) and collagen tpye I level were tested using human dermal fibroblasts (HDFs). Reverse-phase liquid chromatography (RPLC) and ultra-performance liquid chromatography tandem mass spectroscopy (UPLC/MS/MS) were used to quantitatively analyze flavonoids and terpene trilactones in the extraction. The established quantitative analysis method was validated according to the regulatory guidelines. The extraction and compounds kaempferol 3-O-β-D-glucopyranoside, isorhamnetin-3-O-glucoside, myricetin, ginkgolide A and bilobalide could have potential anti-aging activities of G. biloba. The results suggest that G. biloba could be used as anti-aging products and as a cosmetic and medicinal raw material.摘要：本研究旨在测定银杏叶提取物的体外抗衰活性，并鉴定其中的活性化学成分。采用1,1-二苯基-2-三硝基苯肼自由基 (1,1-diphenyl-2-picrylhydrazyl radical, DPPH) 清除实验与2,2'-联氮-双(3-乙基苯并噻唑啉-6-磺酸)自由基 (2,2’-azino-bis (3-ethylbenzothiazoline)-6 sulfonic acid radical, ABTS) 清除实验，评估银杏叶提取物的抗氧化性能；通过人真皮成纤维细胞 (human dermal fibroblasts, HDFs) 模型，测试提取物与活性化学成分对活性氧 (reactive oxygen species)、基质金属蛋白酶-1 (matrix metalloproteinase-1, MMP-1) 以及I型胶原蛋白水平的抑制作用；采用反相液相色谱 (reverse-phase liquid chromatography, RPLC) 与超高效液相色谱串联质谱 (ultra-performance liquid chromatography tandem mass spectroscopy, UPLC/MS/MS)，对提取物中的黄酮类与萜类三内酯类成分进行定量分析，并依据监管指南对所建立的定量分析方法进行方法学验证。本研究发现，银杏提取物以及山柰酚-3-O-β-D-吡喃葡萄糖苷、异鼠李素-3-O-葡萄糖苷、杨梅素、银杏内酯A与白果内酯等化合物，可能是银杏中具备潜在抗衰活性的物质基础。研究结果表明，银杏可用于开发抗衰产品，亦可作为化妆品与医药原料。