水杨叶酚类物质含量及其免疫调节、抗炎、抗癌和抗菌活性

Eman G. Haggag, Mohamed I.S. Abdelhady, Amel M. Kamal
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引用次数: 9

摘要

目的从水杨树叶中分离酚类物质,并评价其免疫调节、抗炎、抗癌和抗菌活性。方法对80%的MeOH叶提取物进行色谱分离,通过不同的色谱和光谱技术(UV、MS、1H和13C NMR)建立化合物的结构。MTT法检测RAW 264.7巨噬细胞增殖的免疫调节作用。分别通过双霉素酸法和ELISA法检测lps刺激细胞对NO和TNF-α因子生成的抑制作用。MTT法测定Hep-G2、MCF-7和HCT-116细胞株的细胞毒性。琼脂扩散法测定抗菌活性。结果首次从该属植物中分离到6个已知的酚类化合物,包括5个类黄酮苷类化合物;芦丁3、槲皮素3- o -阿拉伯葡萄糖苷4、芹菜素7- o -β-d-葡萄糖苷5、槲皮素3- o -α-l-阿拉伯葡萄糖苷6、异槲皮素7及4种糖苷元;山柰素8、芹菜素9、槲皮素10、柚皮素11和两种酚酸;咖啡因和没食子。化合物5对巨噬细胞增殖的促进作用最大,表明其具有免疫调节作用。80% MeOH提取物、4、5和11对lps刺激细胞中亚硝酸盐的抑制作用分别为68.19%、52.95%、20.33%和15.22%,对TNF-α的抑制作用分别为70.82%、29.88%、13.13%和6.14%。80% MeOH叶提取物和化合物4、5对肝细胞癌(Hep-G2)、乳腺癌(MCF-7)和结肠癌(HCT-116)均无细胞毒活性,化合物11仅对HCT-116细胞有细胞毒活性(IC50 = 27.67 μg/ml)。80% MeOH叶提取物对G +ve和G−ve菌均有抑菌活性,且对肺炎克雷伯菌生长有抑制作用,该菌对环丙沙星广谱抗生素具有耐药性。水杨花含有免疫调节、抗炎、细胞毒性和抗菌活性的酚类物质,具有天然药用价值。
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Phenolic content of Ruprechtia salicifolia leaf and its immunomodulatory, anti-inflammatory, anticancer and antibacterial activity

Objectives

This work aimed to isolate phenolics from leaves of Ruprechtia salicifolia and evaluate its immunomodulatory, anti-inflammatory, anticancer and antibacterial activity.

Methods

80% MeOH leaf extract was subjected to chromatographic separation, structures of the isolated compounds were established by different chromatographic and spectral techniques UV, MS, 1H and 13C NMR. Immunomodulatory was evaluated through RAW 264.7 macrophages proliferation by MTT assay. Anti-inflammatory was evaluated by inhibition of NO and TNF-α factor generation in LPS-stimulated cells through bicinchoninici acid assay and ELISA assay, respectively. Cytotoxicity estimated through Hep-G2, MCF-7 and HCT-116 cell lines measured by MTT assay. Antibacterial activity tested by agar diffusion method.

Results

Eleven known phenolic compounds were isolated for the first time from this species including five flavonoid glycosides viz; Rutin 3, quercetin 3-O-arabinoglucoside 4, apigenin 7-O-β-d-glucoside 5, quercetin 3-O-α-l-arabinofuranoside 6 and isoquercetin 7 along with four aglycones viz; kaemferide 8, apigenin 9, quercetin 10 and naringenin 11 and two phenolic acids; caffeic 1 and gallic 2. Compounds 5 showed the most activity increasing macrophage proliferation implying immunomodulatory activity. 80% MeOH extract, 4, 5 and 11 inhibited nitrite oxide by 68.19%, 52.95%, 20.33% and 15.22%, respectively and TNF-α generation by 70.82%, 29.88%, 13.13% and 6.14%, respectively in LPS-stimulated cells implying anti-inflammatory activity. 80% MeOH leaf extract and the tested compounds 4 and 5 were safe possessing no cytotoxic activity against hepatocellular carcinoma (Hep-G2), breast adenocarcinoma (MCF-7) and colon carcinoma (HCT-116), while Compound 11 had cytotoxicity against only HCT-116 cells (IC50 = 27.67 μg/ml). Also 80% MeOH leaf extract showed antibacterial activity against both G +ve and G −ve bacteria, moreover it inhibits growth of Klebsiella pneumonia strain, which is resistant to Ciprofloxacin broad-spectrum antibiotic.

Conclusions

R. salicifolia contain phenolics of immunomodulatory anti-inflammatory, cytotoxicity and antibacterial activity, giving R. salicifolia grate potential as a medicinal natural drug.

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