桑黄水提物中小分子化合物对小鼠肝癌细胞Hepa 1-6的抑制活性分析

Analysis of Anticancer Activity on Mouse Hepatocarcinoma Cells Hepa 1-6 of Small Molecule Compounds in Sanghuangporus spp.Aqueous Extract

桑黄(Sanghuangporus spp.)水提物中多糖、萜烯类化合物等生物大分子具有抑制肿瘤细胞的作用,但桑黄水提物中小分子化合物的功能尚不明确。本研究通过3000 D超滤法获得桑黄小分子化合物,通过双荧光标记法研究其对小鼠肝癌细胞Hepa 1-6的抑制活性,通过代谢组分析法解析其抑癌活性成分。结果表明:桑树桑黄(Sanghuangporus sanghuang Sheng H.Wu,L.W.Zhou&Y.C.Dai)和杨树桑黄(Phellinus baumii)小分子化合物均能诱导小鼠肝癌细胞Hepa 1-6的损伤和凋亡,其抑制率分别为(38.64±4.35)%和(32.52±3.57)%;桑黄小分子化合物与桑黄粗多糖共同处理Hepa 1-6细胞,其抑制率则分别达到(77.27±6.21)%和(76.92±5.45)%;代谢组分析表明桑黄小分子化合物中包括玫瑰树碱、左旋水苏碱、脑苷-B等已知明确具有抑癌功能的化合物,同时鉴定出杂环类(6-氟吲哚、2,4,6-三甲基吡啶、1'-萘酰基-2-甲基吲哚等)、核苷-碱基类(1-甲基腺苷、N6-甲基腺苷、2'-O-甲基腺苷等)、二肽类(如亮氨酸+*、异亮氨酸+*和γ-谷氨酰+*)、糖醇类(木糖醇、核糖醇、甘露醇、赤藓糖醇等)等可能参与肿瘤抑制作用的活性物质。研究表明桑黄水提物含有多种抑制肿瘤细胞的小分子化合物,且与桑黄粗多糖等生物大分子起到协同抑癌作用。研究结果为解析桑黄抑制肿瘤作用机制和功能物质开发应用提供科学依据。

The biological macromolecules such as polysaccharides and terpenes in Sanghuangporus spp.aqueous extract have inhibitory effects on tumor cells,but the function was not yet clear.This study obtained small molecule compounds of Sanghuangporus sanghuang Sheng H.Wu,L.W.Zhou&Y.C.Dai and Phellinus baumii aqueous extracts through 3000 D ultrafiltration,and studied the inhibitory activities on mouse hepatocarcinoma cells Hepa 1-6 using dual fluorescence labeling.The anticancer active ingredients were analyzed using metabolomics analysis.The results showed that the small molecule compounds of S.sanghuang Sheng H.Wu and P.baumii could induce injury and apoptosis of Hepa 1-6,with inhibition rates of(38.64±4.35)%and(32.52±3.57)%,respectively.Meanwhile,the inhibition rates of small molecule compounds reached(77.27±6.21)%and(76.92±5.45)%,respectively,after mixing with crude polysaccharides.Metabolomic analysis showed that anti-cancer small molecular agents such as ellipticine,betonicine,cerebroside-B etc.,were identified in the aqueous extract of Sanghuangporus spp.Antitumor related substances such as heterocyclic compounds(6-fluoroindole,2,4,6-trimethylpyridine,1'-naphthoyl-2-methylindole,etc.),nucleoside and bases(1-methyladenosine,N6-methyladenosine,2'-O-methyladenosine,etc.),dipeptides(leucine+*,isoleucine+*,γ-glutamine+*,etc.),sugar alco-hols(xylitol,ribose,mannitol,erythritol,etc.),were identified too.The results indicate that the aqueous extract of Sanghuangporus spp.contains multiple small molecular agents against tumor cells,which have a synergistic anti-cancer effect with biological macromolecules such as polysaccharides.This would provide a scientific basis for understanding the mechanism of anti-tumor of Sanghuangporus spp.and the development and application of functional substances.