丹参中三萜酸的分离及其抗炎活性的研究
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摘要
目的:对丹参中三萜酸化学成分分离和鉴定及其抗炎活性进行研究。方法:采用减压硅胶柱和凝胶sephadex LH-20柱层析法和中压制备和半制备液相法进行丹参中三萜酸化学成分分离,利用1H-NMR和13C-NMR对分离化合物进行结构鉴定;运用LPS刺激RAW264.7细胞体外抗炎模型对分离化合物进行抗炎活性研究。结果:从丹参中分离得到7个三萜酸,分别鉴定为:(1)2α,3α-dihydroxyolean-12-en-28-oic acid;(2)Maslinicacid;(3) Psiguanin A;(4) 2α,3α-dihydroxyurs-12-en-28-oic acid;(5) 2α-hydroxy ursolic acid;(6) 2α,3α-dihydroxyursa-12,20 (30)-dien-28-oic acid;(7)2α,3α,23-trihydroxyurs-12,20 (30)-dien-28-oic acid 7个化合物都具有较好的抗炎活性,他们的IC50分别是35.6,36.6,40.1,37.8,34.8,32.8,32.1μM,且化合物在高剂量的情况下都能很好的抑制TNF-α和IL-6炎症因子的释放。结论:化合物1、3、6、7都属于该植物首次分离得到,且七个化合物具有一定的抗炎活性。
Objective: To isolate triterpenoid acids from Radix Salviae Miltiorrhizae and evaluate their anti-inflammatory activity. Methods: The reduced-pressure chromatography, sephadex LH-20 chromatography, MPLC, and semi-preparative HPLC were employed to isolate triterpenoids from Radix Salviae Miltiorrhizae, whose chemical structures were identified by1 H-NMR and13 C-NMR. The anti-inflammatory activities were investigated in LPS-stimulated RAW264.7 cells.Results: A total of 7 triterpenoid acids were isolated and identified from Radix Salviae Miltiorrhizae, which were listed as:1) 2α,3α-dihydroxyolean-12-en-28-oic acid, 2) Maslinic acid, 3) Psiguanin A, 4) 2α,3α-dihydroxyurs-12-en-28-oic acid, 5) 2α-hydroxy ursolic acid, 6) 2α,3α-dihydroxyursa-12, 20(30)-dien-28-oic acid, 7) and 2α,3α,23-trihydroxyurs-12,20(30)-dien-28-oic acid. They displayed significant anti-inflammatory activities, whose IC50 were calculated as 35.6, 36.6, 40.1, 37.8, 34.8, 32.8, and 32.1 μM respectively. Using ELISA, we founded that those compounds with high concentration significantly inhibited TNF-α and IL-6 release in LPS-induced RAW264.7 cells.Conclusion: A total of 4 triterpenoid acids such as 1, 3, 6, and 7 are obtained from Radix Salviae Miltiorrhizae for the first time, which exhibit anti-inflammatory activities to a certain degree.
Objective: To isolate triterpenoid acids from Radix Salviae Miltiorrhizae and evaluate their anti-inflammatory activity. Methods: The reduced-pressure chromatography, sephadex LH-20 chromatography, MPLC, and semi-preparative HPLC were employed to isolate triterpenoids from Radix Salviae Miltiorrhizae, whose chemical structures were identified by1 H-NMR and13 C-NMR. The anti-inflammatory activities were investigated in LPS-stimulated RAW264.7 cells.Results: A total of 7 triterpenoid acids were isolated and identified from Radix Salviae Miltiorrhizae, which were listed as:1) 2α,3α-dihydroxyolean-12-en-28-oic acid, 2) Maslinic acid, 3) Psiguanin A, 4) 2α,3α-dihydroxyurs-12-en-28-oic acid, 5) 2α-hydroxy ursolic acid, 6) 2α,3α-dihydroxyursa-12, 20(30)-dien-28-oic acid, 7) and 2α,3α,23-trihydroxyurs-12,20(30)-dien-28-oic acid. They displayed significant anti-inflammatory activities, whose IC50 were calculated as 35.6, 36.6, 40.1, 37.8, 34.8, 32.8, and 32.1 μM respectively. Using ELISA, we founded that those compounds with high concentration significantly inhibited TNF-α and IL-6 release in LPS-induced RAW264.7 cells.Conclusion: A total of 4 triterpenoid acids such as 1, 3, 6, and 7 are obtained from Radix Salviae Miltiorrhizae for the first time, which exhibit anti-inflammatory activities to a certain degree.
引文
1张伯礼,高秀梅,商洪才,等.复方丹参方的药效物质及作用机理研究.世界科学技术-中医药现代化,2003,5(5):14-17.
2刘慧颖,姜长涛,冯娟,等.丹参酮类化合物研究进展.中国药理学通报,2016,2016(12):1643-1647.
3王艳梅,曹俊岭.丹参中酚酸类化合物的化学和药理研究进展.世界中医药,2016,11(6):1126-1130.
4林超,刘兆国,钱星,等.丹酚酸B在心血管疾病中药理作用研究进展.中国药理学通报,2015,31(4):449-452.
5高玉琪,梁春,吴宗贵.丹参水溶性有效成分抗动脉粥样硬化的研究进展.国际中医中药杂志,2012,34(4):364-367.
6陶善珺,任尤楠,赵梦秋,等.丹酚酸B对糖尿病血糖波动模型大鼠胰岛细胞的保护作用.中草药,2016,47(17):3058-3063.
7黄双盛,吴肋杰.丹参的抗氧化与抗炎作用研究进展.中国中医药信息杂志,2002,9(1):86-87.
8李翔,吴磊宏,范骁辉,等.复方丹参方主要活性成分网络药理学研究.中国中药杂志,2011,36(21):2911-2915.
9 Gao H,Huang L,Ding F,et al.Simultaneous purification of dihydrotanshinone,tanshinone I,cryptotanshinone,and tanshinone IIA from Salvia miltiorrhiza and their anti-inflammatory activities investigation.Sci Rep,2018,8(1):8460.
10梁勇,羊裔明.丹参酮药理作用及临床应用研究进展.中草药,2000,31(4):304-306.
11郝文慧,赵文文,陈修平.丹参酮类抗肿瘤作用与机制研究进展.中国药理学通报,2014,30(8):1041-1044.
12易成.丹参酮抗动脉粥样硬化分子机制研究.海峡药学,2011,23(6):43-44.
13刘士林.丹参酮保护心血管疾病的作用机制进展.中国医药科学,2015,5(4):32-34.
14 Wu Y B,Ni Z Y,Shi Q W,et al.Constituents from Salvia species and their biological activities.Chem Rev,2012,112(11):5967-6026.
15 Wang M,Wu F,Chen Y.Studies on the triterpenes from Bemeuxia thibetica Decne.Yao Xue Xue Bao,1993,28(11):845-848.
16 Taniguchi S,Imayoshi Y,Kobayashi E,et al.Production of bioactive triterpenes by Eriobotrya japonica calli.Phytochemistry,2002,59(3):315-323.
17 Shao M,Wang Y,Huang X J,et al.Four new triterpenoids from the leaves of Psidium guajava.J Asian Nat Prod Res,2012,14(4):348-354.
18 Kojima H,Tominaga H,Sato S,et al.Pentacyclic triterpenoids from Prunella vulgaris.Phytochemistry,1987,26(4):1107-1111.
19 Na M K,Yang S,He L,et al.Inhibition of protein tyrosine phosphatase1B by ursane-type triterpenes isolated from Symplocos paniculata.Planta Med,2006,72(3):261-263.
20 Ikuta A,Tomiyasu H,Morita Y,et al.Ursane-and Oleanane-Type Triterpenes from Ternstroemia gymnanthera Callus Tissues.J Nat Prod,2003,66(8):1051-1054.
21 Sashida Y,Ogawa K,Mori N,et al.Triterpenoids from the fruit galls of Actinidia polygama.Phytochemistry,1992,31(8):2801-2804.
22 Gao H,Sun W,Zhao J,et al.Tanshinones and diethyl blechnics with anti-inflammatory and anti-cancer activities from Salvia miltiorrhiza Bunge(Danshen).Sci Rep,2016,6:33720.
23 Stumpf C,Fan Q,Hintermann C,et al.Anti-inflammatory effects of danshen on human vascular endothelial cells in culture.Am J Chin Med,2013,41(5):1065-1077.
24 Lin T H,Hsieh C L.Pharmacological effects of Salvia miltiorrhiza(Danshen)on cerebral infarction.Chin Med,2010,5(1):22.
25 Mennaka K B,Ramesh A,Thomas B,et al.Estimation of nitric oxide as an inflammatory marker in periodontitis.J Indian Soc Periodontol,2009,13(2):75-78.
26 Kwon N S,Nathan C F,Gilker C,et al.L-citrulline production from L-arginine by macrophage nitric oxide synthase.The ureido oxygen derives from dioxygen.J Biol Chem,1990,265:13442-13445.
27 Michael T,Feron O.Nitric oxide synthesis:Which,where,how and why?J Clin Invest,1997,100:2146-2152.
2刘慧颖,姜长涛,冯娟,等.丹参酮类化合物研究进展.中国药理学通报,2016,2016(12):1643-1647.
3王艳梅,曹俊岭.丹参中酚酸类化合物的化学和药理研究进展.世界中医药,2016,11(6):1126-1130.
4林超,刘兆国,钱星,等.丹酚酸B在心血管疾病中药理作用研究进展.中国药理学通报,2015,31(4):449-452.
5高玉琪,梁春,吴宗贵.丹参水溶性有效成分抗动脉粥样硬化的研究进展.国际中医中药杂志,2012,34(4):364-367.
6陶善珺,任尤楠,赵梦秋,等.丹酚酸B对糖尿病血糖波动模型大鼠胰岛细胞的保护作用.中草药,2016,47(17):3058-3063.
7黄双盛,吴肋杰.丹参的抗氧化与抗炎作用研究进展.中国中医药信息杂志,2002,9(1):86-87.
8李翔,吴磊宏,范骁辉,等.复方丹参方主要活性成分网络药理学研究.中国中药杂志,2011,36(21):2911-2915.
9 Gao H,Huang L,Ding F,et al.Simultaneous purification of dihydrotanshinone,tanshinone I,cryptotanshinone,and tanshinone IIA from Salvia miltiorrhiza and their anti-inflammatory activities investigation.Sci Rep,2018,8(1):8460.
10梁勇,羊裔明.丹参酮药理作用及临床应用研究进展.中草药,2000,31(4):304-306.
11郝文慧,赵文文,陈修平.丹参酮类抗肿瘤作用与机制研究进展.中国药理学通报,2014,30(8):1041-1044.
12易成.丹参酮抗动脉粥样硬化分子机制研究.海峡药学,2011,23(6):43-44.
13刘士林.丹参酮保护心血管疾病的作用机制进展.中国医药科学,2015,5(4):32-34.
14 Wu Y B,Ni Z Y,Shi Q W,et al.Constituents from Salvia species and their biological activities.Chem Rev,2012,112(11):5967-6026.
15 Wang M,Wu F,Chen Y.Studies on the triterpenes from Bemeuxia thibetica Decne.Yao Xue Xue Bao,1993,28(11):845-848.
16 Taniguchi S,Imayoshi Y,Kobayashi E,et al.Production of bioactive triterpenes by Eriobotrya japonica calli.Phytochemistry,2002,59(3):315-323.
17 Shao M,Wang Y,Huang X J,et al.Four new triterpenoids from the leaves of Psidium guajava.J Asian Nat Prod Res,2012,14(4):348-354.
18 Kojima H,Tominaga H,Sato S,et al.Pentacyclic triterpenoids from Prunella vulgaris.Phytochemistry,1987,26(4):1107-1111.
19 Na M K,Yang S,He L,et al.Inhibition of protein tyrosine phosphatase1B by ursane-type triterpenes isolated from Symplocos paniculata.Planta Med,2006,72(3):261-263.
20 Ikuta A,Tomiyasu H,Morita Y,et al.Ursane-and Oleanane-Type Triterpenes from Ternstroemia gymnanthera Callus Tissues.J Nat Prod,2003,66(8):1051-1054.
21 Sashida Y,Ogawa K,Mori N,et al.Triterpenoids from the fruit galls of Actinidia polygama.Phytochemistry,1992,31(8):2801-2804.
22 Gao H,Sun W,Zhao J,et al.Tanshinones and diethyl blechnics with anti-inflammatory and anti-cancer activities from Salvia miltiorrhiza Bunge(Danshen).Sci Rep,2016,6:33720.
23 Stumpf C,Fan Q,Hintermann C,et al.Anti-inflammatory effects of danshen on human vascular endothelial cells in culture.Am J Chin Med,2013,41(5):1065-1077.
24 Lin T H,Hsieh C L.Pharmacological effects of Salvia miltiorrhiza(Danshen)on cerebral infarction.Chin Med,2010,5(1):22.
25 Mennaka K B,Ramesh A,Thomas B,et al.Estimation of nitric oxide as an inflammatory marker in periodontitis.J Indian Soc Periodontol,2009,13(2):75-78.
26 Kwon N S,Nathan C F,Gilker C,et al.L-citrulline production from L-arginine by macrophage nitric oxide synthase.The ureido oxygen derives from dioxygen.J Biol Chem,1990,265:13442-13445.
27 Michael T,Feron O.Nitric oxide synthesis:Which,where,how and why?J Clin Invest,1997,100:2146-2152.