仿野生种植三叶青不同部位总黄酮分析及其抗炎、抗氧化能力比较

汪传宝; 陈静文; 王可; 仇凤梅; 黄真; 钟晓明

doi:10.13386/j.issn1002-03062023040159

仿野生种植三叶青不同部位总黄酮分析及其抗炎、抗氧化能力比较

Analysis of Total Flavonoids in Different Parts of Wild Planting Tetrastigma hemsleyanum Diels et Gilg and Comparison of Their Anti-inflammatory and Antioxidant Capacity

摘要

摘要: 目的：比较仿野生种植三叶青不同部位中总黄酮含量、抗氧化活性以及抗炎能力差异。方法：采用醇提工艺对仿野生种植三叶青茎叶、块根、根须进行提取，比较不同部位间总黄酮含量差异；通过DPPH自由基、ABTS⁺自由基、羟自由基清除实验以及铁离子还原力测定，比较仿野生种植三叶青不同部位间抗氧化能力的强弱；采用脂多糖（LPS）诱导RAW264.7细胞作为炎症模型，通过细胞NO的释放量比较仿野生种植三叶青不同部位间的抗炎能力差异。结果：在最佳提取工艺条件下，测得仿野生种植三叶青茎叶、块根、根须总黄酮含量分别为11.86±0.23、8.48±0.10、7.52±0.02 mg·g⁻¹；建立三叶青HPLC指纹图谱，共标定了10个共有峰，通过对照品指认出6号峰为芦丁、8号峰为槲皮素、9号峰为山奈酚，并对其含量进行检测，结果显示块根中三种指标成分含量均最高，其次根须，茎叶最低。检测仿野生种植三叶青不同部位对DPPH自由基、ABTS⁺自由基以及羟自由基的IC₅₀值，茎叶分别为0.2107、0.2315、0.7625 mg/mL；块根分别为0.3134、0.3758、0.8967 mg/mL；根须分别为0.2058、0.2587、0.7856 mg/mL。三叶青茎叶、块根、根须对铁离子还原能力的吸光度值分别为0.172、0.153、0.184。当仿野生种植三叶青三部位的质量浓度在25~200 μg/mL之间时，它们不会对RAW264.7细胞造成毒性，能够有效地抑制LPS诱导的NO释放，从而达到良好的抗炎效果。结论：本研究结果为仿野生种植三叶青不同部位的质量评价提供参考，并对仿野生种植三叶青非药用部位废物再利用及后续全资源开发提供理论依据。

Abstract: Objective: The content of total flavonoids, antioxidant activity and anti-inflammatory ability in different parts of Tetrastigma hemsleyanum Diels et Gilg which imitated wild planting were compared. Methods: Stems, leaves, root tubers and root whiskers of wild imitating-planted Tetrastigma hemsleyanum Diels et Gilg were first extracted by alcohol extraction technology, and the content of total flavonoids was compared among different parts. Then DPPH free radical scavenging rate, ABTS⁺ free radical scavenging rate, hydroxyl free radical scavenging rate and ferric ion reducing power were used to evaluate the antioxidant capacity of different parts of plants. Meanwhile, lipopolysaccharide (LPS)-induced RAW264.7 cell inflammation model was established to compare anti-inflammatory ability of different parts of wild planting Tetrastigma hemsleyanum Diels et Gilg by measuring the release of NO. Results: Under the optimal extraction conditions, the contents of total flavonoids in stem leaves, root tubers and root whiskers were 11.86±0.23, 8.48±0.10 and 7.52±0.02 mg·g⁻¹, respectively. A total of 10 common peaks of Tetrastigma hemsleyanum Diels et Gilg were identified by HPLC fingerprint. Rutin (peak 6), quercetin (peak 8) and kaempferol (peak 9) were identified via comparisons with reference standards. The content of the three indexes was the highest in roots, followed by stems and leaves. The IC₅₀ values of DPPH free radicals, ABTS⁺ free radicals and hydroxyl free radicals in different parts of the plant were detected, and the values of stem leaves were 0.2107, 0.2315 and 0.7625 mg/mL. Root tubers were 0.3134, 0.3758 and 0.8967 mg/mL. The roots were 0.2058, 0.2587 and 0.7856 mg/mL, respectively. The absorbance values of the reducing capacity of ferric ions in stem, tuber and root were 0.172, 0.153 and 0.184. Three parts of the wild imitating-wild plant were not toxic to RAW264.7 cells at the concentrations between 25~200 μg/mL and could effectively inhibit the release of NO induced by LPS that achieving a good anti-inflammatory effect. Conclusions: The results of this study provided a reference for the quality evaluation and a theoretical basis for waste recycling as well as subsequent whole resource development to the non-medicinal parts of Tetrastigma hemsleyanum Diels et Gilg from the wild.