木豆种子和荚的化学成分及其抗氧化、抗炎活性

doi:10.11707/j.1001-7488.LYKX20220286

摘要/Abstract

摘要：

目的: 对木豆种子和荚部位的化学成分进行分离鉴定，并研究这些化学成分的抗氧化、抗炎活性，以期为发现新的天然抗氧化、抗炎活性成分以及木豆资源的开发利用提供科学依据。方法: 分别采用95％乙醇回流提取木豆种子和荚，提取液过滤后减压浓缩得到木豆种子和荚的浸膏。浸膏分别用不同有机溶剂萃取，对乙酸乙酯萃取部继续采用硅胶、凝胶等柱层析色谱分离技术进行进一步的分离、纯化。利用MS、NMR等技术鉴定分离得到的化合物的结构。通过测定化合物DPPH自由基清除能力评价化合物的抗氧化活性，采用Griess法测定化合物对脂多糖诱导小鼠巨噬细胞RAW 264.7产生NO的抑制能力评价化合物的抗炎活性。结果: 从木豆种子乙酸乙酯萃取部中分离出4个化合物，通过ESI-MS、NMR等技术鉴定为对苯二酚（S1）、1–亚油酰–3–棕榈酰甘油（S2）、胡萝卜苷亚油酸酯（S3）、豆甾烷–3β,6α–二醇（S4）。从木豆荚乙酸乙酯萃取部中分离出5个化合物，根据理化性质初步鉴定为黄酮类化合物，通过ESI-MS、NMR等技术鉴定为2',5,7–三羟基–4'–甲氧基异黄酮（P1）、毛蕊异黄酮（P2）、大豆苷元（P3）、2'-羟基染料木素（P4）、染料木素（P5）。化合物S1~S4、P1~P2为首次从木豆属植物中分离得到，其中化合物S2为新化合物。抗氧化和抗炎活性研究表明，化合物S1、P1、P2、P4具有较强的DPPH自由基清除活性，EC₅₀值分别达到（63.10±0.15）、（64.97±0.67）、（79.63±1.14）和（79.43±1.29） μmol·L^?1，化合物S1~S2、P1~P5具有较强的NO抑制活性，IC₅₀值分别为（31.36±0.44）、（52.08±1.11）、（30.66±0.83）、（46.30±0.90）、（55.18±2.07）、（44.11±0.79）和（44.92±2.24） μmol·L^?1。化合物P1的抗氧化和抗炎活性以及化合物P4的抗炎活性为首次报道。结论: 木豆种子中主要含脂肪酸及甾醇衍生物，抗氧化和抗炎活性较弱，木豆荚中含有的多种异黄酮类成分具有较好的抗氧化和抗炎活性。因此，木豆荚更具有开发利用价值，可作为天然抗氧化及抗炎成分的提取原料来源。

关键词: 木豆种子, 荚, 化学成分, 异黄酮, 抗氧化活性, 抗炎活性

Abstract:

Objective: In this study, the chemical constituents of Cajanus cajan seeds and pods were separated and identified, as well as their antioxidant and anti-inflammatory activities were investigated, in order to provide a scientific basis for the discovery of new natural antioxidant and anti-inflammatory active constituents and the development and utilization of C. cajan resources. Method: The seeds and pods of C. cajan were extracted by 95% ethanol reflux, respectively, and the extracts were filtered and concentrated by reduced pressure to obtain the seed and pod residues. The residues were extracted with different organic solvents, and the ethyl acetate extraction section was further separated and purified by column chromatography techniques such as silica gel and gel. The structures of the isolated compounds were identified using MS and NMR techniques. The antioxidant activities of the isolated compounds were evaluated by measuring the scavenging capacities of DPPH free radicals. The anti-inflammatory activities of the isolated compounds were evaluated by Griess method for determining the inhibitory abilities of the compounds on the NO production in mouse macrophage RAW 264.7 induced by LPS. Result: Four compounds were isolated from the ethyl acetate extraction of C. cajan seeds and identified by ESI-MS and NMR techniques as hydroquinone (S1), 1-linoleoyl-3-palmitoyl glycerol (S2), daucosterol linoleate (S3) and stigmastane-3β,6α-diol (S4). Five compounds were isolated from the ethyl acetate extraction of C. cajan pods and preliminarily identified as flavonoids based on their physicochemical properties, and further identified by ESI-MS and NMR techniques as 2',5,7-trihydroxy-4'-methoxyisoflavone (P1), calycosin (P2), daidzein (P3), 2'-hydroxygenistein (P4) and genistein (P5). Compounds S1–S4, P1–P2 were isolated from the genus Cajanus for the first time, while compound S2 was a new compound. Antioxidant and anti-inflammatory activities studies showed that compounds S1, P1, P2, P4 had relatively strong DPPH free radical scavenging activities, with the EC₅₀ values of 63.10 ± 0.15, 64.97 ± 0.67, 79.63 ± 1.14 and 79.43 ± 1.29 μmol·L^?1, respectively, while compounds S1–S2, P1–P5 showed strong NO inhibitory activities, with the IC₅₀ values of 31.36±0.44, 52.08±1.11, 30.66±0.83, 46.30±0.90, 55.18±2.07, 44.11±0.79 and 44.92±2.24 μmol·L^?1, respectively. The antioxidant and anti-inflammatory activities of compound P1 and the anti-inflammatory activity of compound P4 were reported for the first time. Conclusion: C. cajan seeds mainly contain fatty acid and sterol derivatives, with relatively weak antioxidant and anti-inflammatory activities, while C. cajan pods contain multiple isoflavones with better antioxidant and anti-inflammatory activities. Therefore, C. cajan pods are more valuable for development and utilization, and can be used as a source of raw materials for extracting natural antioxidant and anti-inflammatory components.

Key words: Cajanus cajan seed, pod, chemical constituent, isoflavone, antioxidant activity, anti-inflammatory activity

中图分类号:

Q946.8

徐晓俞,李程勋,李爱萍. 木豆种子和荚的化学成分及其抗氧化、抗炎活性[J]. 林业科学, 2023, 59(2): 67-74.

Xiaoyu Xu,Chengxun Li,Aiping Li. Chemical Constituents of Cajanus cajan Seeds and Pods and Their Antioxidant and Anti-Inflammatory Activities[J]. Scientia Silvae Sinicae, 2023, 59(2): 67-74.

图/表 4

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No.	δ_H	δ_C
1	4.20 (1H, dd, J = 11.6, 4.6 Hz)4.15 (1H, dd, J = 11.7, 6.2 Hz)	65.32
2	3.93 (1H, m)	70.41
3	3.69 (1H, dd, J = 11.4, 3.9 Hz)3.60 (1H, dd, J = 11.4, 5.8 Hz)	63.47
1′	–	174.51
2′	2.35 (2H, t, J = 7.5 Hz)	34.31
3′	1.63 (2H, m)	25.06
4′–7′, 15′	1.25~1.36^a	29.24~29.84
8′	2.05 (2H, dd, J = 13.8, 6.8 Hz)	27.35
9′	5.39 (1H, d, J = 6.3 Hz)	130.15
10′	5.32 (1H, m)	128.22
11′	2.77 (2H, t, J = 6.6 Hz)	25.78
12′	5.34 (1H, m)	128.03
13′	5.37 (1H, d, J = 6.6 Hz)	130.38
14′	2.05 (2H, dd, J = 13.8, 6.8 Hz)	27.33
16′	1.25~1.36^a	31.67
17′	1.25~1.36^a	22.84
18′	0.89 (3H, t, J = 6.6 Hz)	14.27
1″	–	174.46
2″	2.35 (2H, t, J = 7.5 Hz)	34.29
3″	1.63 (2H, m)	25.04
4″–13″	1.25~1.36^a	29.24~29.84
14″	1.25~1.36^a	32.07
15″	1.25~1.36^a	22.72
16″	0.87 (3H, t, J = 7.0 Hz)	14.22

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