Cytotoxic Activity of Ethyl Acetate Fraction of Aglaia elliptica Blume Leaves Extract on HepG2 Hepatocarcinoma Cells

Churiyah Churiyah, Ekclesia Yuliarni, A. E. Wibowo, Firdayani Firdayani
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Abstract

Aglaia elliptica Blume belongs to Meliaceae family which contain an active compound of rocaglamid as anticancer. The research was conducted to evaluate the biological activities of ethyl acetate fraction of Aglaia elliptica Blume leaves. The dry powder of Aglaia elliptica Blume leaves was extracted with methanol using maceration method, then was fractionated using n-hexane and ethyl acetate, guided by brine shrimp lethality test (BSLT). The most toxic fraction then processed by column chromatography and resulted in eight subfractions. Three of them showed the most toxic effect on BSLT namely FEA 3.3, FEA 3.4 and FEA 3.5 with LC50 of 40.81, 18.56 and 13.40 ppm respectively. The cytotoxic activities of those three active subfractions were assayed on HepG2 hepatocarcinoma cells using enzymatic reaction of 3-(4,5-dimethylthiazoyl-2-yl) 2,5 diphenyltetrazolium bromide (MTT), and the results showed that the IC50 were 35.10, 14.36 and 14.09 ppm respectively. The most active sub fraction (FEA 3.5) was then performed further analyzed using preparative high performance liquid chromatography (HPLC). The HPLC results indicated that there were three active compounds, which were suspected as derivatives of rocaglamid. The molecular docking simulation indicated that rocaglamide formed complex with Toll-like Receptor 4 in HepG2 hepatocarcinoma cells and affected the inhibition of proliferation of its cell. Keywords : Aglaia elliptica Blume leaves extract, fractionation, BSLT, MTT assay,hepatocarsinoma cell lines
乌桕叶提取物乙酸乙酯部分对HepG2肝癌细胞的细胞毒活性
白菖蒲属楝科植物,含有一种具有抗癌作用的活性化合物。本研究对白菖蒲叶乙酸乙酯部位的生物活性进行了评价。采用甲醇浸渍法提取黑木莲叶片干粉,用正己烷和乙酸乙酯进行分馏,以卤虾致死试验(BSLT)为指导。然后用柱层析法对毒性最强的部分进行处理,得到8个亚部分。其中FEA 3.3、FEA 3.4和FEA 3.5对BSLT的毒性作用最大,LC50分别为40.81、18.56和13.40 ppm。用3-(4,5-二甲基噻唑-2-基)2,5二苯基溴化四唑(MTT)酶促反应测定了3种活性亚组分对HepG2肝癌细胞的细胞毒活性,结果表明IC50分别为35.10、14.36和14.09 ppm。然后用制备型高效液相色谱法(HPLC)进一步分析最活性亚组分(FEA 3.5)。高效液相色谱分析结果表明,其中有3个活性化合物可能为罗卡酰胺衍生物。分子对接模拟表明,rocaglamide在HepG2肝癌细胞中与toll样受体4形成复合物,影响其细胞增殖抑制作用。关键词:黑木莲叶提取物,分离,BSLT, MTT法,肝癌细胞系
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