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采用气质联用分析结合多元分析方法研究产自印度的四种姜黄属植物根茎精油的挥发性成分及其体外抗肿瘤和抗炎活性。

Volatile profiling coupled with multivariate analysis, antiproliferative and anti-inflammatory activities of rhizome essential oil of four Hedychium species from India.

机构信息

Centre for Biotechnology, Siksha 'O' Anusandhan (Deemed to be University), Bhubaneswar, Odisha, India.

Centre for Biotechnology, Siksha 'O' Anusandhan (Deemed to be University), Bhubaneswar, Odisha, India.

出版信息

J Ethnopharmacol. 2023 Dec 5;317:116835. doi: 10.1016/j.jep.2023.116835. Epub 2023 Jun 22.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

The genus Hedychium of family Zingiberaceae comprises several perennial rhizomatous species widely used in perfumery and traditional folk medicine to treat diseases related to asthma, diarrhoea, nausea, stomach disorders, inflammation and tumours. Several species of Hedychium have remained under-explored with respect to their chemical composition and biological activities.

AIM OF THE STUDY

The current research aimed to explore the chemical composition and evaluate the antiproliferative and anti-inflammatory activities of rhizome essential oil from four Hedychium species (H. coccineum, H. gardnerianum, H. greenii and H. griffithianum).

MATERIALS AND METHODS

Compound identification was accomplished using a Clarus 580 gas chromatography system in conjunction with mass spectrometry (GC-MS). The multivariate data statistics using chemometrics (PCA, PLS-DA, sPLS-DA) and cluster analysis (Dendrogram, Heat maps, K-means) were used to compare the similarity and relationship among Hedychium metabolomes. MTT assay was employed to visualize the antiproliferative property against MCF7, HepG2 and PC3 cancerous cell lines. The toxicity of essential oils was determined using 3T3-L1 non-tumorigenic/normal cells. Lipopolysaccharide (LPS)-induced RAW 264.7 cells were used to investigate the anti-inflammatory properties of Hedychium essential oils by measuring the production of nitric oxide (NO) using the Griess reagent method. Furthermore, the levels of prostaglandin (PGE) and pro-inflammatory cytokines (TNF-α, IL-6, IL-1β) was assessed using the ELISA technique.

RESULTS

In total, 82 compounds were identified in four targeted species of Hedychium from which 1,8-cineole (52.71%), β-pinene (32.83%), α-pinene (19.62%), humulene epoxide II (19.86%) and humulene epoxide I (19.10%) were the major constituents. Monoterpenes (8.5-59.9%) and sesquiterpenes (2.87-54.11%) were the two class of compounds, found as the most prevalent in the extracted essential oils. The multivariate analysis classified the four Hedychium species into three different clusters. Hedychium essential oils exhibited potent antiproliferative activity against MCF7, HepG2 and PC3 cancer cell lines with IC values less than 150 μg/mL where H. gardnerianum exhibited the highest selective cytotoxicity against human breast and prostate adenocarcinoma cells with an IC value of 44.04 ± 1.07 μg/mL and 56.11 ± 1.44 μg/mL, respectively. The essential oils on normal (3T3-L1) cells displayed no toxicity with higher IC values thereby concluding as safe to use for normal human health without causing any side effects. Besides, the essential oils at 100 μg/mL concentration revealed remarkable anti-inflammatory activity in LPS-activated RAW 264.7 murine macrophages by inhibiting the production of inflammatory mediators, with H. greenii exhibiting the maximum anti-inflammation response by significantly suppressing the levels of NO (84%), PGE (87%), TNF-α (94.3%), IL-6 (95%) and IL-1β (85%) as compared to LPS treated group.

CONCLUSION

The present findings revealed that the Hedychium species traditionally used in therapeutics could be a potential source of active compounds with antiproliferative and anti-inflammatory properties.

摘要

植物化学分类学意义

姜科姜黄属植物包括多种多年生根茎植物,广泛用于香水和传统民间医学,用于治疗与哮喘、腹泻、恶心、胃部疾病、炎症和肿瘤相关的疾病。姜黄属的几种植物在化学成分和生物活性方面仍然研究不足。

研究目的

本研究旨在探索四种姜黄属植物(红姜黄、绿姜黄、云南姜黄和金佛山姜黄)根茎精油的化学成分,并评估其抗增殖和抗炎活性。

材料和方法

使用 Clarus 580 气相色谱系统与质谱联用(GC-MS)进行化合物鉴定。使用多元数据分析方法(主成分分析(PCA)、偏最小二乘判别分析(PLS-DA)、偏最小二乘判别分析(sPLS-DA))和聚类分析(树状图、热图、K-均值)比较姜黄属代谢组之间的相似性和关系。MTT 法用于可视化 MCF7、HepG2 和 PC3 癌细胞系的抗增殖特性。使用 3T3-L1 非肿瘤/正常细胞测定精油的毒性。使用脂多糖(LPS)诱导的 RAW 264.7 细胞,通过格里斯试剂法测量一氧化氮(NO)的产生来研究姜黄属精油的抗炎特性。此外,通过 ELISA 技术评估前列腺素(PGE)和促炎细胞因子(TNF-α、IL-6、IL-1β)的水平。

结果

共鉴定出四种姜黄属植物中的 82 种化合物,其中 1,8-桉叶素(52.71%)、β-蒎烯(32.83%)、α-蒎烯(19.62%)、葎草烯环氧化物 II(19.86%)和葎草烯环氧化物 I(19.10%)为主要成分。单萜(8.5-59.9%)和倍半萜(2.87-54.11%)是两种最常见的化合物类型,存在于提取的精油中。多变量分析将四种姜黄属植物分为三个不同的聚类。姜黄属精油对 MCF7、HepG2 和 PC3 癌细胞系具有明显的抗增殖活性,IC 值小于 150μg/mL,其中 H. gardnerianum 对人乳腺癌和前列腺腺癌细胞的选择性细胞毒性最高,IC 值分别为 44.04±1.07μg/mL 和 56.11±1.44μg/mL。精油对正常(3T3-L1)细胞没有毒性,IC 值较高,因此可以安全用于人类健康,不会产生任何副作用。此外,在 LPS 激活的 RAW 264.7 鼠巨噬细胞中,精油在 100μg/mL 浓度下具有显著的抗炎活性,通过抑制炎症介质的产生来抑制炎症,其中 H. greenii 通过显著抑制 NO(84%)、PGE(87%)、TNF-α(94.3%)、IL-6(95%)和 IL-1β(85%)的水平,表现出最大的抗炎反应与 LPS 处理组相比。

结论

本研究结果表明,传统用于治疗的姜黄属植物可能是具有抗增殖和抗炎活性的活性化合物的潜在来源。

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