药学学报

C No.	δ_H (J/Hz)	δ_C
1′	-	169.7, C
2′	5.71 (1H, s)	119.2, CH
3′	-	156.7, C
4′	2.52 (2H, t, J = 6.4)	40.6, CH₂
5′	4.39 (1H, m); 4.28 (1H, m)	63.1, CH₂
1	-	175.8, C
2	4.10 (1H, d, J = 3.6)	74.3, CH
3	1.77 (1H, m)	39.9, CH
4	1.48 (1H, m); 1.27 (1H, m)	27.0, CH₂
5	0.94 (3H, overlap)	12.1, CH₃
6	0.84 (3H, d, J = 6.7)	14.1, CH₃
6′	2.17 (3H, d, J = 1.1)	18.5, CH₃

C No.	δ_H (J/Hz)	δ_C
1′	-	169.7, C
2′	5.71 (1H, s)	119.2, CH
3′	-	156.7, C
4′	2.52 (2H, t, J = 6.4)	40.6, CH₂
5′	4.39 (1H, m); 4.28 (1H, m)	63.1, CH₂
1	-	175.8, C
2	4.10 (1H, d, J = 3.6)	74.3, CH
3	1.77 (1H, m)	39.9, CH
4	1.48 (1H, m); 1.27 (1H, m)	27.0, CH₂
5	0.94 (3H, overlap)	12.1, CH₃
6	0.84 (3H, d, J = 6.7)	14.1, CH₃
6′	2.17 (3H, d, J = 1.1)	18.5, CH₃

溶洞真菌Furcasterigmium sp. L-YB-P1-1的次级代谢产物研究

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田静源 , 舒婷 , 李金严 , 位彩霞 , 孔凡栋 , 王佩 , 许海棠 , 周丽曼 ^*

药学学报 | 研究论文 2026,61(7): 2590-2595

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药学学报 |研究论文 2026 , 61 (7) : 2590 -2595

溶洞真菌Furcasterigmium sp. L-YB-P1-1的次级代谢产物研究

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田静源, 舒婷, 李金严, 位彩霞, 孔凡栋, 王佩, 许海棠, 周丽曼^*

作者信息

广西民族大学化学化工学院, 林产化学与工程国家民委重点实验室, 广西林产化学与工程重点实验室, 林木生物质低碳高质利用广西高校工程研究中心, 广西南宁 530006

通讯作者:

周丽曼, E-mail: zhouliman88@126.com

作者简介:

作者贡献:

田静源是本文第一作者, 负责实验设计与论文的写作; 舒婷负责论文的修改; 李金严负责化合物的分离纯化; 位彩霞负责样本的采集和前处理; 孔凡栋负责实验指导及结构解析的指导; 王佩、许海棠负责提供了研究所需的经费及关键资源; 周丽曼是本文的通讯作者, 负责实验设计及把关, 提供本文思路, 参与稿件修改等工作。

Study on secondary metabolites of cave-derived fungus Furcasterigmium sp. L-YB-P1-1

Jing-yuan TIAN, Ting SHU, Jin-yan LI, Cai-xia WEI, Fan-dong KONG, Pei WANG, Hai-tang XU, Li-man ZHOU^*

Affiliations

Key Laboratory of Chemistry and Engineering of Forest Products, State Ethnic Affairs Commission, Guangxi Key Laboratory of Chemistry and Engineering of Forest Products, Engineering Research Center of Low-Carbon and High-Quality Utilization of Forest Biomass, University of Guangxi; School of Chemistry and Chemical Engineering, Guangxi Minzu University, Nanning 530006, China

出版时间: 2026-07-12 doi: 10.16438/j.0513-4870.2026-0040

文章导航

摘要

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对源自广西金伦洞的真菌Furcasterigmium sp. L-YB-P1-1的次级代谢产物粗提物进行提取研究。利用薄层色谱、高效液相色谱等对真菌次级代谢产物粗提物进行分离纯化, 并运用现代波谱技术HNMR、质谱等测定化合物结构, 得到10个化合物, 其中包括1个新化合物(2R,3S)-2-hydroxy-3-methylpentanoic acid (E)-5′-hydroxy-3′-methylpent-2′-enoate (1)和9个已知化合物: uridine (2)、adenosine (3)、2-methoxyl-β-L-arabinofuroside uracil (4)、thymidine (5)、2′-O-methyladenosine (6)、5′-O-acetyl-9-β-arabinofuranosyl adenosine (7)、uridine-5′-α-hydroxypropanoate (8)、asperteretal G (9)及trans-ferulic acid (10)。使用DPPH法对化合物进行抗氧化活性评价, 结果显示化合物2~5和10具有抗氧化活性; 使用Griess法检测化合物对LPS诱导RAW264.7细胞产生NO的含量的影响, 发现化合物9具有一定的抗炎活性。

关键词

溶洞真菌 / Furcasterigmium sp. L-YB-P1-1 / 次级代谢产物 / 结构鉴定 / 活性评价

Abstract

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The secondary metabolites of the fungus Furcasterigmium sp. L-YB-P1-1, isolated from Jinlun Cave in Guangxi were investigated. The crude extract was separated and purified by chromatographic techniques including thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). The structures of the isolated compounds were elucidated through comprehensive spectroscopic analysis such as HNMR and mass spectrometry. Ten compounds were obtained, comprising one new compound, (2R,3S)-2-hydroxy-3-methylpentanoic acid (E)-5′-hydroxy-3′-methylpent-2′-enoate (1) and 9 known compounds: Uridine (2), adenosine (3), 2-methoxyl-β-L-arabinofuranosyl uracil (4), thymidine (5), 2′-O-methyladenosine (6), 5′-O-acetyl-9-β-arabinofuranosyl adenosine (7), uridine-5′-α-hydroxypropanoate (8), asperteretal G (9), and trans-ferulic acid (10). The antioxidant activity of the compounds was evaluated using the DPPH free radical scavenging assay, which revealed that compounds 2-5 and 10 possessed antioxidant activity. Furthermore, the anti-inflammatory activity was assessed by measuring the inhibition of NO production in LPS-induced RAW 264.7 cells using the Griess method, indicating that compound 9 exhibited certain anti-inflammatory activity.

Key words

Cave fungus / Furcasterigmium sp. L-YB-P1-1 / secondary metabolites / structural identification / activity evaluation

引用本文

田静源, 舒婷, 李金严, 位彩霞, 孔凡栋, 王佩, 许海棠, 周丽曼. 溶洞真菌Furcasterigmium sp. L-YB-P1-1的次级代谢产物研究. 药学学报, 2026 , 61 (7) : 2590 -2595 . DOI: 10.16438/j.0513-4870.2026-0040

Jing-yuan TIAN, Ting SHU, Jin-yan LI, Cai-xia WEI, Fan-dong KONG, Pei WANG, Hai-tang XU, Li-man ZHOU. Study on secondary metabolites of cave-derived fungus Furcasterigmium sp. L-YB-P1-1[J]. Acta Pharmaceutica Sinica, 2026 , 61 (7) : 2590 -2595 . DOI: 10.16438/j.0513-4870.2026-0040

正文

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溶洞作为一种独特的生态环境, 具有光照稀缺、温度波动小、湿度高以及营养贫瘠等特点。这种长期与外界相对隔离的环境, 促进了具有特殊代谢途径的真菌类群的生长与演化。根据现有研究, 溶洞中蕴藏着丰富的真菌资源^[1-4], 目前已报道的真菌物种约2 000种, 分属于青霉属(Penicillium)、链格孢属(Alternaria)、枝孢菌属(Cladosporium)等多个类群。然而, 相较于物种多样性调查, 针对溶洞真菌次级代谢产物的研究尚处于起步阶段, 相关报道仍较少。已有研究表明, 溶洞真菌能产生结构新颖且具有多种生物活性的次级代谢产物, 例如从溶洞真菌中分离到的代谢产物显示出抗肿瘤^{[5, 6]}、抗氧化^[7]、促血管生成^[8]等生物活性。此外, 真菌在共培养等策略下能被激活产生通常条件下沉默的代谢产物, 后续还可通过共培养等策略进一步挖掘这些菌株的代谢潜力。因此, 探索溶洞真菌的次级代谢产物, 对于发现结构新颖的先导化合物、开发新型药物以及理解真菌的生态适应机制均具有重要意义, 展现出巨大的潜力。

在Furcasterigmium sp. L-YB-P1-1中共分离得到10个化合物, 其中包括1个新化合物(2R,3S)-2-hydroxy-3-methylpentanoic acid (E)-5′-hydroxy-3′-methylpent-2′-enoate (1)和9个已知化合物: uridine (2)、adenosine (3)、2-methoxyl-β-L-arabinofuroside uracil (4)、thymidine (5)、2′-O-methyladenosine (6)、5′-O-acetyl-9-β-arabinofuranosyl adenosine (7)、uridine-5′-α-hydroxypropanoate (8)、asperteretal G (9)及trans-ferulic acid (10) (图1)。抗氧化活性评价结果显示, 从菌株Furcasterigmium sp. L-YB-P1-1中分离鉴定的化合物2~5和10具有一定的抗氧化活性, 此外化合物9具有一定的抗炎活性。

结果与讨论

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1　菌株鉴定

菌株L-YB-P1-1 ITS序列通过PCR扩增获得长度为536 bp序列, 通过检索NCBI数据库并进行基因序列对比, 该真菌菌株鉴定为Furcasterigmium sp.属, GenBank登录号为PX908945, 该菌株的系统发育树如图2所示。

2　结构解析

化合物1为无色油状物, 其分子式通过正离子高分辨电喷雾质谱(HRESIMS)确定为C₁₂H₂₀O₅, 比旋光度[α]²⁵_D = 12 (c 0.100 0, CH₃OH)。化合物1的¹H NMR和¹³C NMR (表1)光谱显示有3个甲基基团[δ_C/H (18.5/2.17, d, J = 1.1 Hz, CH₃-6′), δ_C/H (14.1/0.84, d, J = 6.7 Hz, CH₃-6), δ_C/H (12.1/0.94, overlap, CH₃-5)]; 3个亚甲基[δ_C/H (63.1/4.39, m, CH₂-5′; 63.1/4.28, m, CH₂-5′), δ_C/H (40.6/2.52, t, J = 6.4 Hz, CH₂-4′), δ_C/H (27.0/1.48, m, CH₂-4; 27.0/1.27, m, CH₂-4)]; 3个次甲基[δ_C/H (74.3/4.10, d, J = 3.6 Hz, CH-2), δ_C/H (39.9/1.77, m, CH-3)], 包括一个sp²次甲基[δ_C/H (119.2/5.71, s, CH-2′)]; 一个羧基碳, 一个sp²杂化双键碳和一个酯基碳。上述数据表明该化合物具有三个不饱和度。根据¹H-¹H COSY图谱, 显示了H-4′/H-5′、H-2/H-3/H-4/H-5以及H-3/H-6之间的相关性。HMBC图显示H-2′与C-1′、C-4′相连, H-5′与C-1相连, H-2与C-1相连, H-3与C-1相连, H-6′与C-2′、C-3′、C-4′相连。通过对HMBC和COSY数据的详细分析, 确定了该化合物的平面结构(图3)。为明确C-2和C-3的构型, 分别对两种可能的构型(2R,3S)-1与(2R,3R)-1进行了¹³C NMR化学位移的理论计算, 并将计算结果与实验数据进行了系统比较。DP4+概率分析结果显示, (2R,3S)-1构型与实验13C NMR数据具有更高的匹配概率。在此基础上, 进一步对该构型进行了ECD计算。结果表明, (2R,3S)-1构型不仅其理论 ¹³C NMR化学位移与实验值吻合良好, 其计算的ECD曲线也与实验测得的ECD谱图高度一致(图4), 因此确定C-2和C-3的绝对构型分别为R型和S型。因此, 化合物1被命名为(2R,3S)-2-hydroxy-3-methylpentanoic acid (E)-5′-hydroxy-3′-methylpent-2′-enoate, 即(2R,3S)-2-羟基-3-甲基戊酸(E)-5′-羟基-3′-甲基戊-2′-烯酸酯。

3　抗氧化活性

化合物的抗氧化活性通过DPPH法进行测定, 实验数据表明, 当化合物浓度为20 μmol·L^-1时, 化合物2~5和10的DPPH自由基清除率分别为73.61% ± 4.36%、62.50% ± 6.26%、68.06% ± 7.25%、47.22% ± 5.67%、59.72% ± 6.13%, 阳性对照(V_C)的DPPH自由基清除率为88.89% ± 5.21%。

4　抗炎活性

对Furcasterigmium sp. L-YB-P1-1中获得的含量较大的次级代谢产物化合物2、3和9进行抗炎活性测试, 实验数据表明化合物9在20、10 µmol·L^-1浓度下均具有一定的抗炎活性, 对炎症因子NO的产生具有显著的抑制作用。

实验部分

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WZZ-2S数字旋光仪; Brucker AVANCE 400 MHz核磁共振(德国Brucker公司); 日立Primaide高效液相色谱仪(北京日立科学仪器有限公司); 分析高效色谱柱C₁₈-MS-Ⅱ (C18, 250 mm × 4.6 mm, 5 µm)、5PFP (50 mm × 4.6 mm, 5 µm), 日本Nacalai tesque公司; 兰博seriesⅢ制备型高效液相色谱, 制备型高效液相色谱柱5C18-MS-Ⅱ (C₁₈, 250 mm × 10 mm, 5 µm); 5PFP (250 mm × 10 mm, 5 µm), 日本Nacalai tesque公司。

1　菌株的发酵

Furcasterigmium sp. L-YB-P1-1来源于广西金伦洞穴的岩壁附着物, 该附着物是溶洞中经水溶蚀和再沉积形成的碳酸钙沉积物。纯化后的菌株通过ITS rDNA基因序列鉴定。本实验使用马铃薯培养基: 葡萄糖2%, 琼脂2%, 马铃薯20%; 大米培养基: 大米80 g, 水120 mL。Furcasterigmium sp. L-YB-P1-1菌株在28 ℃的马铃薯葡萄糖琼脂培养基上纯化培养7 d, 将培养的菌株分别接入含碳酸氢钙溶液和不含碳酸氢钙溶液的培养基中, 在室温下发酵培养30 d。

2　化合物的提取和分离

发酵30 d后, Furcasterigmium sp. L-YB-P1-1的发酵产物用乙酸乙酯在室温减压下萃取, 得到41.8 g粗提取物。粗提物进一步用90%甲醇-水混合液和石油醚按1∶1的比例萃取, 得到14.3 g甲醇层提取物和30.4 g石油醚层提取物。甲醇层提取物经正相硅胶柱色谱, 以石油醚-乙酸乙酯(10∶1~1∶2)梯度洗脱, 得到26个馏分, 分别为Fr.1~Fr.26。馏分Fr.5 (646.7 mg)经C₁₈高速制备型HPLC处理, 得到5个亚馏分, 即Fr.5.1~Fr.5.5。Fr.5.4 (85.8 mg)经半制备型HPLC (50%甲醇-酸水-C₁₈柱)进一步纯化, 得到化合物1 (6.4 mg)和化合物9 (31.7 mg)。馏分Fr.26 (1.0 g)经半制备型HPLC (10%甲醇-水-C18柱)纯化, 得到化合物2 (7.4 mg)和化合物3 (21.9 mg)。馏分Fr.25 (512.0 mg)经C₁₈高速制备型HPLC处理, 得到12个亚馏分, 即Fr.25.1~Fr.25.12。Fr.25.1被鉴定为化合物8 (5.7 mg), Fr.25.4 (9.3 mg)经半制备型HPLC (15%甲醇-水-5PFP柱)进一步纯化, 得到化合物4 (2.1 mg)。Fr.25.5 (19.7 mg)经半制备型HPLC (15%甲醇-水-5PFP柱)纯化, 得到化合物5 (8.5 mg)。Fr.25.6 (9.3 mg)经半制备型HPLC (20%甲醇-水-5PFP柱)纯化, 得到化合物6 (1.8 mg)。Fr.25.11 (5.5 mg)经半制备型HPLC (30%甲醇-水-5PFP柱)纯化, 得到化合物7 (1.4 mg)。馏分Fr.11经半制备型HPLC (40%甲醇-水-C₁₈柱)纯化, 得到化合物10 (2.9 mg)。

3　结构鉴定

化合物2: 黄色油状; ESI-MS m/z 267.0 [M+Na]⁺, 分子式C₉H₁₂N₂O₆; ¹H NMR (400 MHz, CD₃OD): δ_H 8.02 (1H, d, J = 7.9 Hz, H-6), 5.91 (1H, d, J = 4.6 Hz, H-7), 5.70 (1H, d, J = 8.2 Hz, H-5), 4.16 (2H, m, H-8, H-9), 4.01 (1H, s, H-10), 3.84 (1H, dd, J = 2.5, 12.3 Hz, H-11a), 3.73 (1H, dd, J = 3.1, 12.2 Hz, H-11b); ¹³C NMR (100 MHz, CD₃OD): δ_C 166.2 (C, C-4), 152.4 (C, C-2), 142.7 (CH, C-6), 102.6 (CH, C-5), 90.6 (CH, C-7), 86.4 (CH, C-10), 75.7 (CH, C-8), 71.3 (CH, C-9), 62.4 (CH₂, C-11)。该化合物的数据与文献^[9]报道的一致, 故化合物2鉴定为uridine。

化合物3: 白色粉末; ESI-MS m/z 268.1 [M+H]⁺, 分子式C₁₀H₁₃N₅O₄; ¹H NMR (400 MHz, CD₃OD): δ_H 8.32 (1H, s, H-2), 8.18 (1H, s, H-8), 5.96 (1H, d, J = 6.3 Hz, H-1′), 4.74 (1H, dd, J = 4.9, 6.3 Hz, H-2′), 4.32 (1H, dd, J = 2.5, 5.1 Hz, H-3′), 4.17 (1H, dd, J = 2.5, 5.0 Hz, H-4′), 3.89 (1H, dd, J = 2.5, 12.5 Hz, H-5′a), 3.74 (1H, dd, J = 2.5, 12.5 Hz, H-5′b); ¹³C NMR (100 MHz, CD₃OD): δ_C 157.6 (C, C-6), 153.5 (CH, C-3), 150.0 (C, C-4), 142.0 (CH, C-5), 121.0 (C, C-5), 91.3 (CH, C-1′), 88.2 (CH, C-4′), 75.6 (CH, C-2′), 72.7 (CH, C-3′), 63.5 (CH₂, C-5′)。该化合物的数据与文献^{[10, 11]}报道的一致, 故化合物3鉴定为adenosine。

化合物4: 无色油状; ESI-MS m/z 259.1 [M+H]⁺, 分子式C₁₀H₁₄N₂O₆; ¹H NMR (400 MHz, CD₃OD): δ_H 8.09 (1H, d, J = 8.1 Hz, H-6), 5.94 (1H, d, J = 3.6 Hz, H-1′), 5.69 (1H, d, J = 8.1 Hz, H-5), 4.24 (1H, t, J = 5.8 Hz, H-4′), 3.98 (1H, m, H-3′), 3.86 (2H, overlap, H-2′, H-5′a), 3.75 (1H, dd, J = 3.2, 12.3 Hz, H-5′b), 3.52 (3H, s, 2′-OMe); ¹³C NMR (100 MHz, CD₃OD): δ_C 166.3 (C, C-4), 152.2 (C, C-2), 142.5 (CH, C-6), 102.5 (CH, C-5), 88.8 (CH, C-1′), 86.1 (CH, C-3′), 85.0 (CH, C-2′), 69.7 (CH, C-4′), 61.6 (CH₂, C-5′), 58.8 (CH₃, 2′-OMe)。该化合物的数据与文献^[12]报道的一致, 故化合物4鉴定为2-methoxyl-β-L-arabinofuroside uracil。

化合物5: 白色油状; ESI-MS m/z 243.1 [M+H]⁺, 分子式C₁₀H₁₄N₂O₅; ¹H NMR (400 MHz, CD₃OD): δ_H 7.82 (1H, d, J = 1.3 Hz, H-2), 6.28 (1H, t, J = 7.0 Hz, H-1′), 4.40 (1H, m, H-4′), 3.91 (1H, dd, J = 3.5, 6.7 Hz, H-3′), 3.80 (1H, dd, J = 3.1, 12.0 Hz, H-5′a), 3.73 (1H, dd, J = 3.7, 12.0 Hz, H-5′b), 2.23 (2H, m, H-2′), 1.88 (3H, d, J = 1.2 Hz, H-7); ¹³C NMR (100 MHz, CD₃OD): δ_C 166.5 (C, C-4), 152.4 (C, C-6), 138.2 (CH, C-2), 111.6 (C, C-3), 88.8 (CH, C-1′), 86.2 (CH, C-4′), 72.2 (CH, C-3′), 62.8 (CH₂, C-5′), 41.2 (CH₂, C-2′), 12.5(CH₃, C-7)。该化合物的数据与文献^[13]报道的一致, 故化合物5鉴定为thymidine。

化合物6: 白色固体; ESI-MS m/z 304.1 [M+Na]⁺, 分子式C₁₁H₁₅N₅O₄; ¹H NMR (400 MHz, CD₃OD): δ_H 8.37 (1H, s, H-7), 8.21 (1H, s, H-2), 6.08 (1H, d, J = 5.9 Hz, H-1′), 4.49 (1H, dd, J = 3.0, 5.0 Hz, H-3′), 4.42 (1H, t, J = 6.0 Hz, H-2′), 4.16 (1H, dd, J = 2.8, 5.6 Hz, H-4′), 3.89 (1H, dd, J = 2.5, 12.2 Hz, H-5′a), 3.76 (1H, dd, J = 2.8, 12.2 Hz, H-5′b), 3.42 (3H, s, 2′-OMe); ¹³C NMR (100 MHz, CD₃OD): δ_C 152.9 (CH, C-7), 142.0 (CH, C-2), 89.2 (CH, C-1′), 88.4 (CH, C-4′), 84.7 (CH, C-2′), 70.8 (CH, C-3′), 63.2 (CH₂, C-5′), 58.8 (CH₃, 2′-OMe)。该化合物的数据与文献^{[14, 15]}报道的一致, 故化合物6鉴定为2′-O-methyladenosine。

化合物7: 白色固体; ESI-MS m/z 332.1 [M+Na]⁺, 分子式C₁₂H₁₅N₅O₅; ¹H NMR (400 MHz, CD₃OD): δ_H 8.32 (1H, s, H-2), 8.26 (1H, s, H-7), 6.03 (1H, d, J = 4.5 Hz, H-1′), 4.73 (1H, t, J = 4.8 Hz, H-5′a), 4.34~4.40 (3H, overlap, H-5′b, H-2′, H-3′), 4.26 (1H, t, J = 5.0 Hz, H-4′), 2.06 (3H, s, H-7′); ¹³C NMR (100 MHz, CD₃OD): δ_C 172.4 (C, C-6′), 169.6 (C, C-4), 152.0 (C, C-9), 141.8 (CH, C-7), 90.5 (CH, C-1′), 83.6 (CH, C-4′), 75.2 (CH, C-2′), 71.8 (CH, C-3′), 64.9 (CH₂, C-5′), 20.7 (CH₃, C-7′)。该化合物的数据与文献^[16]报道的一致, 故化合物7鉴定为5′-O-acetyl-9-β-arabinofuranosyl adenosine。

化合物8: 白色粉末; ESI-MS m/z 339.0 [M+Na]⁺, 分子式C₁₂H₁₆N₂O₈; ¹H NMR (400 MHz, CD₃OD): δ_H 7.83 (1H, d, J = 8.1 Hz, H-6), 5.87 (1H, d, J = 4.2 Hz, H-1′), 5.71 (1H, d, J = 8.2 Hz, H-5), 4.42 (1H, dd, J = 3.9, 12.4 Hz, H-5′a), 4.37 (1H, d, J = 2.7 Hz, H-5′b), 4.33 (2H, dd, J = 3.1, 10.0 Hz, H-2′, H-2″), 4.20 (2H, overlap, H-4′, H-3′), 1.40 (3H, d, J = 6.9 Hz, H-3″); ¹³C NMR (100 MHz, CD₃OD): δ_C 175.9 (C, C-1″), 166.2 (C, C-4), 152.3 (C, C-2), 142.7 (CH, C-6), 102.9 (CH, C-5), 91.3 (CH, C-1′), 82.9 (CH, C-4′), 71.5 (CH, C-2′), 71.2 (CH, C-3′), 68.1 (CH, C-2′′), 65.0 (CH₂, C-5′), 20.7 (CH₃, C-3″)。该化合物的数据与文献^[17]报道的一致, 故化合物8鉴定为uridine-5′-α-hydroxypropanoate。

化合物9: 白色粉末; ESI-MS m/z 291.1 [M+Na]⁺, 分子式C₁₆H₁₂O₄; ¹H NMR (400 MHz, CD₃OD): δ_H 7.62 (2H, d, J = 7.3 Hz, H-2″, H-6″), 7.27 (2H, t, J = 7.3 Hz, H-3″, H-5″), 7.22 (1H, d, J = 7.3 Hz, H-4″), 7.16 (2H, d, J = 8.4 Hz, H-2′, H-6′), 6.74 (2H, d, J = 8.5 Hz, H-3′, H-5′), 6.23 (1H, s, H-4); ¹³C NMR (100 MHz, CD₃OD): δ_C 171.5 (C, C-1), 159.6 (C, C-4′), 140.1 (C, C-2), 132.1 (C, C-3), 130.6 (CH, C-2′, C-6′), 129.5 (C, C-1″), 129.4 (CH, C-4″), 129.3 (CH, C-3″, C-5″), 128.9 (CH, C-2″, C-6″), 128.4 (C, C-1′), 116.7 (CH, C-3′, C-5′), 82.3 (CH, C-4)。该化合物的数据与文献^[18]报道的一致, 故化合物9鉴定为asperteretal G。

化合物10: 黄色粉末; ESI-MS m/z 217.0 [M+Na]⁺, 分子式C₁₀H₁₀O₄; ¹H NMR (400 MHz, CD₃OD): δ_H 7.60 (1H, d, J = 15.9 Hz, H-7), 7.18 (1H, d, J = 2.1 Hz, H-2), 7.06 (1H, dd, J = 1.9, 8.1 Hz, H-6), 6.81 (1H, d, J = 8.2 Hz, H-5), 6.31 (1H, d, J = 16.0 Hz, H-8), 3.89 (3H, s, 3-OMe); ¹³C NMR (100 MHz, CD₃OD): δ_C: 171.0 (C, C-9), 150.5 (C, C-4), 149.4 (C, C-3), 146.9 (CH, C-7), 127.8 (C, C-1), 124.0 (CH, C-6), 116.5 (CH, C-5), 115.9 (CH, C-8), 111.7 (CH, C-2), 56.4 (CH₃, 3-OMe)。该化合物的数据与文献^[19]报道的一致, 故化合物10鉴定为trans-ferulic acid。

4　抗氧化活性

抗氧化活性采用DPPH法进行测定。样品和V_C均配制成甲醇溶液, 浓度为20 µmol·L^-1。DPPH称量后配制成0.15 µmol·L^-1的甲醇溶液, 需在低温避光条件下保存。在96孔板中设置了以下组别, 空白对照组: 200 µL甲醇溶液; 阴性对照组: 160 µL甲醇溶液和40 µL DPPH溶液; V_C对照组: 160 µL V_C溶液和40 µL甲醇溶液; V_C + DPPH组: 160 µL V_C溶液和40 µL DPPH溶液; 样品对照组: 160 µL样品溶液和40 µL甲醇溶液; 样品+ DPPH组: 160 µL样品溶液和40 µL DPPH溶液, 每孔均设置3个重复。将96孔板置于暗处30 min, 使用酶标仪在517 nm波长下测定各孔的吸光度(OD)值, 随后计算清除率。

5　抗炎活性测试

通过测量LPS处理的RAW 264.7细胞中NO的产生来筛选抗炎活性。将RAW 264.7细胞以5×10⁴个细胞/孔的密度在96孔板中培养24 h。用LPS (1 μg·mL^-1)刺激细胞, 然后用不同浓度的样品培养24 h。以吲哚美辛为阳性对照, 取上清液50 μL, 加入Griess试剂(试剂A 50 μL, 试剂B 50 μL), 用酶标仪在540 nm处测定OD值, 在实验过程中, 为每个测量的浓度设置了3个平行。计算化合物对NO产量的抑制率, 将抑制率高于50%的化合物, 进行IC₅₀测试。IC₅₀实验方案浓度配置为0、0.625、1.25、2.5、5、10、20、30、40、50 μmol·L^-1, 实验步骤与初筛一致, 使用IBM SPSS Statistics软件根据其各浓度抑制率计算得IC₅₀值。

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2026年第61卷第7期

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doi: 10.16438/j.0513-4870.2026-0040

接收时间：2026-01-09
首发时间：2026-09-21
出版时间：2026-07-12

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收稿日期：2026-01-09
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周丽曼, E-mail: zhouliman88@126.com

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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