金花茶体外抗肿瘤活性及物质基础的初步研究

Studies on Antitumor Effects of Camellia chrysantha in vitro and its Material Basis

目的:观察金花茶3种不同萃取部位对5种不同人肿瘤细胞株体外增殖的作用,寻找其活性部位及物质基础。方法:采用噻唑蓝染色法(MTT法),检测金花茶不同萃取部位体外对人胃腺癌细胞株(SGC-7901)、人大细胞肺癌细胞株(H460)、人肝癌细胞株(SMMC-7721和BEL-7404)、人高分化鼻咽癌细胞株(CNE-1)作用48 h后的增殖抑制率,并计算相应的半数生长抑制浓度(IC50),其中,细胞密度分别为:SGC-7901(7×104/mL),H460(4×104/mL),SMMC-7721(6×104/mL),BEL-7404(6×104/mL),CNE-1(4×104/mL),不同提取部位的浓度分别为:正丁醇层(50,100,150,200 mg·L-1)、乙酸乙酯层(50,100,200,300 mg·L-1)、水层(25,50,75,100,150 mg·L-1);采用液质联用方法初步分析水层萃取部分的可能活性成分。结果:金花茶正丁醇、乙酸乙酯、水层部位对5种肿瘤均有抑制作用(P<0.05或P<0.01),且呈现一定的浓度依赖性,各部分对5种肿瘤细胞的抑制作用强弱为:水层>正丁醇层>乙酸乙酯层,水层部位对上述5种细胞株的IC50分别为:81.72,73.47,95.98,73.41,61.25 mg·L-1;水层萃取部分检测到的物质有24个,初步鉴定了其中的11个成分[分别为:(1R,3R,4R,5R)-1,3,4,5-tetrahydroxycyclohexanecarboxylic acid;daucic acid;二羟基-6-甲氧基-四氢-2H-吡喃-3-氧基-四氢-2H-吡喃-3,4,5-三醇;coumaroylquinic acid;1,3,4-tri-O-acetyl-alpha-D-fructofuranosyl,3,6-tri-O-acetyl-alpha-D-glucopyranoside;2S,3aR,4'S,5'S,6R,6'R,7R,7aS)-6,6'-bis[(acetyloxy)methyl]tetrahydro-4H,4'H-spiro[1,3-dioxolo[4,5-c]pyran-2,3'-pyran]-3a,4',5',7(6H)-tetrayl tetraacetate;(2S,3R,4R,5R,6S)-2-(2S,3R,4S,5S,6R)-4,5-二羟基-2-(5-羟基-7-甲氧基-2-(4-甲氧苯基)-4-羰基-4H-色烯-6-基)-6-(羟甲基)-四氢-2H-吡喃-3-氧基-4,5-二羟基-6-甲基-四氢-2H-吡喃-3-基乙酸酯;vitexin-7-glucoside;luteolin-7-diglucoside;7-O-β-D-glucopyranosyl-kaempferol-3-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranoside;ilexside II],同时表征了另外13个未鉴定成分的质谱信息。结论:金花茶不同萃取部位体外实验有抗肿瘤活性,水层部分可能是金花茶抗肿瘤作用的主要活性部位;需要进一步对水层部分的这些可能活性物质进行分离纯化鉴定以及定量分析。

Objective： To observe the inhibitory effects of different extractive fractions from Camellia chrysantha on five different kinds of malignant tumor cell lines, and evaluate the antitumor effects in vitro and its material basis of the effects. Method： MTT assay was adopted to detect the inhabitation of different extractive fractions from C. chrysantha with five different kinds of malignant tumor cell lines in vitro and IC50 values were calculated,respectively. The UPLC-LTQ-Orbitrap MS/MS was used to analyse its possible components. Result： The different extractive fractions from C. chrysantha had antitumor effects on SGC-7901, H460, SMMC-7721, BEL-7404, CNE-1,they inhibited cell proliferation in a dose dependent manner. The sequence of their inhabitation intensity was：water-soluble part〉n-butyl alcohol part〉ethyl acetate part, and the IC50 of water-soluble part on the five different kinds cell lines was 81.72,73.47,95.98,73.41,61.25 mg·L^-1,respectively;24 components were investigated, 11 of them were tentatively identified, and 13 of them were characterized by their mass spectra information. Conclusion： Different extractive fractions from C. chrysantha can inhibit the proliferation of SMMC-7721, BEL-7404, CNE-1, SGC-7901 and H460 in vitro,the water-soluble part is considered as antitumor active part of C. chrysantha;but which component exactly contribute to the antitumor effect remains to be further confirmed.