婆婆纳抗黑色素瘤物质基础的初步研究

Preliminary study on the material basis of anti-melanoma activity of Veronica didyma

关键词:  婆婆纳, 化学成分, 黑色素瘤, 分离纯化, 活性导向

Key words:  Veronica didyma Tenore, chemical composition, melanoma, separation and purification, bioassay-guided

摘要:

为明确婆婆纳(Veronica didyma)抗黑色素瘤活性部位及物质基础,该研究采用CCK8法评价了婆婆纳乙醇提取物4个萃取部位(石油醚层、乙酸乙酯层、正丁醇层、水层)乙醇提取物及单体化合物对黑色素瘤细胞株(B16和A375)细胞的增殖抑制作用,并使用植物化学方法和技术对活性部位的化学成分进行系统分离纯化。结果表明:(1)乙酸乙酯萃取部位(ethyl acetate extract,PPNE)较其他样品有更好抑制B16细胞和A375细胞增殖的作用,其半抑制浓度(IC₅₀)值分别为0.177 mg·mL^-1(B16)、2.826 mg·mL^-1(A375)。(2)从活性部位PPNE中得到7个单体化合物,即对羟基苯甲醛(1)、胡黄连苷II(2)、isoscutellarein 7-O-(6-oacetyl)-β-allopyranosyl(1→2″)-β-glucopyranoside(3)、3'-hydroxyl-4'-O-methylisoscutellarein 7-O-[6-O-acetyl-β-D-allopyranosyl-(1→2)-β-D-glucopyranoside(4)、6-O-veratroylcatalposide(5)、veronicoside(6)、 isoscutellarein 4'-methyl ether 7-O-(6-O-acetyl)-ballopyranosyl(1→2″)-β-glucopyranoside(7)。(3)7个化合物均为首次从该植物中分离得到,HPLC分析结果显示这7个化合物是PPNE的较大量成分。(4)除化合物1外,其余6个单体化合物均有较好抑制黑色素瘤细胞增殖的作用,该文首次报道了化合物3、4、7的抗黑色素瘤活性。综上认为,婆婆纳的乙酸乙酯萃取部位(PPNE)为其抗黑色素瘤活性部位,环烯醚萜类化合物2、5、6和黄酮类化合物3、4、7可能为PPNE抗黑色素瘤的活性基础。该研究结果为合理利用其资源奠定了科学基础。

Abstract:

To clarify the anti-melanoma active extract and material basis of Veronica didyma, CCK8 method was used to evaluate the inhibitory effects of four extracts(petroleum ether extract, ethyl acetate extract, n-butanol extract, water extract), ethanol extracts and monomer compounds on the proliferation of melanoma cell lines(B16 and A375). The chemical constituents of the active fraction were systematically separated and purified by phytochemical techniques and methods. The results were as follows:(1)Ethyl acetate extract(PPNE)had a better inhibitory effect on the proliferation of B16 and A375 cells than other samples, and IC₅₀ = 0.177 mg·mL^-1(B16), IC₅₀ = 2.826 mg·mL^-1(A375), respectively.(2)Seven monomer compounds were obtained from PPNE were p-hydroxybenzaldehyde(1), picroside II(2), isoscutellarein 7-O-(6-oacetyl)-β-allopyranosyl(1→2″)-β-glucopyranoside(3), 3'-hydroxyl-4'-O-methylisoscutellarein 7-O-[6-O-acetyl-β-D-allopyranosyl-(1→2)-β-D-glucopyranoside(4), 6-O-veratroylcatalposide(5), veronicoside(6), isoscutellarein 4'-methyl ether 7-O-(6-O-acetyl)-ballopyranosyl(1→2″)-β-glucopyranoside(7).(3)Seven compounds were isolated from this plant for the first time, and HPLC showed that these seven compounds were the major components of PPNE.(4)Except Compound 1, the other six monomers all had good inhibitory effects on the proliferation of melanoma cells, and the anti-melanoma activities of compounds 3, 4 and 7 were reported for the first time. Based on these results, PPNE of V. didyma is an anti-melanoma active fraction, iridoids(compounds 2, 5, 6)and flavonoids(compounds 3, 4, 7)may be the basis of anti-melanoma activity of PPNE. The results of this study will lay scientific foundation for the rational utilization of V. didyma's resources.