三花龙胆化学成分及抗肿瘤活性研究
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摘要
目的探讨三花龙胆(Gentiana triflora Pall.)的化学成分及其体外抗肿瘤活性。方法采用95%乙醇对三花龙胆根茎进行回流提取,依次用石油醚、乙酸乙酯、正丁醇进行萃取,萃取物用硅胶柱色谱、制备型高效液相色谱进行分离纯化,根据理化性质和波谱数据对化合物进行结构鉴定;采用四氮唑盐还原法(MTT)评价各化合物的抗肿瘤活性。结果从乙酸乙酯和正丁醇萃取部位分离得到5个化合物,分别鉴定为globuloside A(1)、cornusoside A(2)、cornolactone A(3)、6,9-epi-8-O-Acetylshanziside methyl ester(4)、5,9-epi-Penstemoside(5);抗肿瘤活性实验表明,化合物1对HepG2细胞的体外增殖具有一定的抑制作用,IC50值为6.7μmol/L。结论所有化合物均首次从该植物中分离得到;化合物1对HepG2细胞的增殖具有明显的抑制活性。
Objective To investigate the chemical constituents and antitumor activities in vitro of Gentiana triflora Pall.Methods The root of Gentiana triflora Pall. was extracted with 95% ethyl alcohol, which is extracted with petroleum ether, ethyl acetate and n-butanol, successively. The extracts were separated and purified by silica gel column chromatography and preparative high performance liquid chromatography. The compounds were structurally identified according to physicochemical properties and spectral data. The antitumor activities of the compounds were evaluated by microculture tetrozolium(MTT) method. Results Five compounds were isolated from the ethyl acetate and n-butanol extraction sites and identified as globuloside A(1), cornusoside A(2), cornolactone A(3), 6, 9-epi-8-O-Acetylshanziside methyl ester( 4), 5,9-epi-Penstemoside(5); anti-tumor activity experiments showed that Compound 1 had a certain inhibitory effect on the proliferation of HepG2 cells in vitro, with an IC50 value of 6.7 μmol/L. Conclusion All the compounds are isolated from Gentiana triflora Pall. firstly. Compound 1 has significant inhibitory activity on the proliferation of HepG2 cells.
Objective To investigate the chemical constituents and antitumor activities in vitro of Gentiana triflora Pall.Methods The root of Gentiana triflora Pall. was extracted with 95% ethyl alcohol, which is extracted with petroleum ether, ethyl acetate and n-butanol, successively. The extracts were separated and purified by silica gel column chromatography and preparative high performance liquid chromatography. The compounds were structurally identified according to physicochemical properties and spectral data. The antitumor activities of the compounds were evaluated by microculture tetrozolium(MTT) method. Results Five compounds were isolated from the ethyl acetate and n-butanol extraction sites and identified as globuloside A(1), cornusoside A(2), cornolactone A(3), 6, 9-epi-8-O-Acetylshanziside methyl ester( 4), 5,9-epi-Penstemoside(5); anti-tumor activity experiments showed that Compound 1 had a certain inhibitory effect on the proliferation of HepG2 cells in vitro, with an IC50 value of 6.7 μmol/L. Conclusion All the compounds are isolated from Gentiana triflora Pall. firstly. Compound 1 has significant inhibitory activity on the proliferation of HepG2 cells.
引文
[1]国家药典委员会.中华人民共和国药典[S].一部.北京:化学工业出版社,2005,64.
[2] Wang S,Xu Y,Jiang W,et al. Isolation and identification of constituents with activity of inhibiting nitric oxide production in RAW 264.7 macrophages from Gentiana triflora[J].Planta Med,2013,79(8):680-686.
[3] Koshioka M,Miyamoto K,Horio T,et al. Identification of endogenous gibberellins in stems and leaves in vegetative growth stage of Gentiana triflora[J]. J Plant Physiol,1998,153(1/2):230-232.
[4] Suyama Y,Tanaka N,Kawazoe K,et al. Rigenolides D-H,norsecoiridoid and secoiridoids from Gentiana rigescens Franch[J]. J Nat Med,2018,72(2):1-6.
[5] Yang Y,Wang Z,Zhang L,et al. Protective effect of gentiopicroside from Gentiana macrophylla Pall. in ethanolinduced gastric mucosal injury in mice[J]. Phytother Res,2017,32(2):259-266.
[6] Xu Y,Li Y,Maffucci KG,et al. Analytical methods of phytochemicals from the genus Gentiana[J]. Molecules,2017,22(12):2080.
[7] Chen B,Peng Y,Wang X,et al. preparative separation and purification of four glycosides from Gentianae radix by high-speed counter-current chromatography and comparison of their anti-NO production effects[J]. Molecules,2017,22(11):2002.
[8] Ghazanfar K,Mubashir K,Dar SA,et al. Gentiana kurroo Royle attenuates the metabolic aberrations in diabetic rats;Swertiamarin,swertisin and lupeol being the possible bioactive principles[J]. J Complement Integr Med,2017,14(3):27.
[9] Zou YF,Fu YP,Chen XF,et al. Polysaccharides with immunomodulating activity from roots of Gentiana crassicaulis[J]. Carbohydr Polym,2017,172:306-314.
[10] Cao X,Guo X,Yang X,et al. Transcriptional Responses and Gentiopicroside Biosynthesis in Methyl JasmonateTreated Gentiana macrophylla Seedlings[J]. PLloS One,2016,11(11):e0 166 493.
[11] Pan Y,Zhao YL,Zhang J,et al. Phytochemistry and Pharmacological Activities of the Genus Gentiana(Gentianaceae)[J]. Chem. Biodivers, 2016,47(16):107.
[12] Waltenberger B,Liu R,Atanasov AG,et al. Nonprenylated Xanthones from Gentiana lutea,Frasera caroliniensis,and Centaurium erythraea as Novel Inhibitors of Vascular Smooth Muscle Cell Proliferation[J]. Molecules,2015,20(11):20 381.
[13] Wang Z,Wang C,Su T,et al. Antioxidant and immunological activities of polysaccharides from Gentiana scabra Bunge roots[J]. Carbohydr Polym,2014,112(112):114-118.
[14] Wang S,Xu Y,Chen P,et al. Structural characterization of secoiridoid glycosides by high-performance liquid chromatography/electrospray ionization mass spectrometry[J].Rapid Commun Mass Spectrom,2014,28(14):1569-79.
[15] Xu M,Wang D,Zhang YJ,et al. Iridoidal glucosides from Gentiana rhodantha[J]. J Asian Nat Prod Res,2008,10(6):491-498.
[16]徐毅敏.三花龙胆化学成分及其抗炎活性研究[D].杭州:浙江大学,2013.
[17] Yang AM,Sun J,Li H,et al. Chemical Constituents and Anti-Tumor Activity of Gentiana farreri[J]. Advan Mat Res,2012,554-556:1682-1685.
[18] Cali s覨I,Kirmizibekmez H,Sticher O. Iridoid glycosides from Globularia trichosantha[J]. J Nat Prod,2001,64(1):60-64.
[19] He Y,Peng J,Hamann M T,et al. An iridoid glucoside and the related aglycones from Cornus florida[J]. J Nat Prod,2014,77(9):2138-43.
[20] Delazar A,Byres M,Gibbons S,et al. Iridoid Glycosides from Eremostachys glabra[J]. J Nat Prod,2004,67(9):1584-1587.
[2] Wang S,Xu Y,Jiang W,et al. Isolation and identification of constituents with activity of inhibiting nitric oxide production in RAW 264.7 macrophages from Gentiana triflora[J].Planta Med,2013,79(8):680-686.
[3] Koshioka M,Miyamoto K,Horio T,et al. Identification of endogenous gibberellins in stems and leaves in vegetative growth stage of Gentiana triflora[J]. J Plant Physiol,1998,153(1/2):230-232.
[4] Suyama Y,Tanaka N,Kawazoe K,et al. Rigenolides D-H,norsecoiridoid and secoiridoids from Gentiana rigescens Franch[J]. J Nat Med,2018,72(2):1-6.
[5] Yang Y,Wang Z,Zhang L,et al. Protective effect of gentiopicroside from Gentiana macrophylla Pall. in ethanolinduced gastric mucosal injury in mice[J]. Phytother Res,2017,32(2):259-266.
[6] Xu Y,Li Y,Maffucci KG,et al. Analytical methods of phytochemicals from the genus Gentiana[J]. Molecules,2017,22(12):2080.
[7] Chen B,Peng Y,Wang X,et al. preparative separation and purification of four glycosides from Gentianae radix by high-speed counter-current chromatography and comparison of their anti-NO production effects[J]. Molecules,2017,22(11):2002.
[8] Ghazanfar K,Mubashir K,Dar SA,et al. Gentiana kurroo Royle attenuates the metabolic aberrations in diabetic rats;Swertiamarin,swertisin and lupeol being the possible bioactive principles[J]. J Complement Integr Med,2017,14(3):27.
[9] Zou YF,Fu YP,Chen XF,et al. Polysaccharides with immunomodulating activity from roots of Gentiana crassicaulis[J]. Carbohydr Polym,2017,172:306-314.
[10] Cao X,Guo X,Yang X,et al. Transcriptional Responses and Gentiopicroside Biosynthesis in Methyl JasmonateTreated Gentiana macrophylla Seedlings[J]. PLloS One,2016,11(11):e0 166 493.
[11] Pan Y,Zhao YL,Zhang J,et al. Phytochemistry and Pharmacological Activities of the Genus Gentiana(Gentianaceae)[J]. Chem. Biodivers, 2016,47(16):107.
[12] Waltenberger B,Liu R,Atanasov AG,et al. Nonprenylated Xanthones from Gentiana lutea,Frasera caroliniensis,and Centaurium erythraea as Novel Inhibitors of Vascular Smooth Muscle Cell Proliferation[J]. Molecules,2015,20(11):20 381.
[13] Wang Z,Wang C,Su T,et al. Antioxidant and immunological activities of polysaccharides from Gentiana scabra Bunge roots[J]. Carbohydr Polym,2014,112(112):114-118.
[14] Wang S,Xu Y,Chen P,et al. Structural characterization of secoiridoid glycosides by high-performance liquid chromatography/electrospray ionization mass spectrometry[J].Rapid Commun Mass Spectrom,2014,28(14):1569-79.
[15] Xu M,Wang D,Zhang YJ,et al. Iridoidal glucosides from Gentiana rhodantha[J]. J Asian Nat Prod Res,2008,10(6):491-498.
[16]徐毅敏.三花龙胆化学成分及其抗炎活性研究[D].杭州:浙江大学,2013.
[17] Yang AM,Sun J,Li H,et al. Chemical Constituents and Anti-Tumor Activity of Gentiana farreri[J]. Advan Mat Res,2012,554-556:1682-1685.
[18] Cali s覨I,Kirmizibekmez H,Sticher O. Iridoid glycosides from Globularia trichosantha[J]. J Nat Prod,2001,64(1):60-64.
[19] He Y,Peng J,Hamann M T,et al. An iridoid glucoside and the related aglycones from Cornus florida[J]. J Nat Prod,2014,77(9):2138-43.
[20] Delazar A,Byres M,Gibbons S,et al. Iridoid Glycosides from Eremostachys glabra[J]. J Nat Prod,2004,67(9):1584-1587.