中药山茱萸的化学成分和生物活性研究

发布时间：2018-06-30 19:16

本文选题：山茱萸 + 化学成分　；参考：《北京中医药大学》2017年硕士论文

【摘要】：山茱萸为山茱萸科(Cornaceae)植物山茱萸Cornus officinalis Sieb.et Zucc.的干燥成熟果肉,主产于陕西、山西、河南等省,是我国传统的名贵滋补中药材。其药用历史已有两千余年,始载于《神农本草经》"治心下邪气寒热,温中逐寒湿痹,去三虫,久服轻身",其性酸、涩,微温,归肝、肾经,具有补益肝肾和涩精固脱之功效。山茱萸是一些经典中药名方的主药,如六味地黄丸、金匮肾气丸、左归丸等。研究表明山茱萸的化学成分主要为环烯醚萜类、鞣质类、黄酮类、三萜类和苯丙素类等。现代药理学研究表明山茱萸在抗炎、糖尿病的防治、心脑血管的保护、神经保护、抗肿瘤、抗氧化和抗衰老等方面均表现出较好的活性。虽然山茱萸含有多种药理活性,但其研究多集中于粗提物水平,而单体化合物少有报道。为了更好的阐明山茱萸的药效物质基础,对其化学成分进行系统的研究具有十分重要的意义。本论文对山茱萸水提物的化学成分进行了较系统地分离和结构鉴定,并且对分离到的单体化合物进行了初步的抗炎和神经保护作用的评价,为山茱萸的进一步开发利用提供了科学依据。根据环烯醚萜类化合物的最大紫外吸收在230 nm处特点,采用化学导向分离的思路,运用硅胶柱色谱、ODS柱色谱、SephadexLH-20柱色谱和制备高效液相色谱等方法从山茱萸中分离纯化得到80个化合物,根据UV、IR、MS、HRESIMS、1DNMR、2DNMR、CD、ECD等技术方法,鉴定了其中72个化合物的结构,包括41个环烯醚萜苷类化合物、3个单萜类化合物、5个黄酮类化合物、13个木脂素类化合物以及10个其它类型化合物(化合物名称见下表),它们分别为:cornuside A(1*),cornuside B(2*),cornuside C(3*),cornuside D(4*),cornuside E(5*),cornuside F(6*),cornuside G(7*),cornuside H(8*),cornuside I(9*),cornuside J(10*),cornuside K(11*),cornuside L(12*),cornuside M(13*),cornuside N(14*),cornuside O(15*),cornuside P(16*),cornuside Q(17*),cornusfuroside A(18*),cornusfuroside B(19*),cornusfuroside C(20*),cornusfuroside D(21*),6'-O-acetyl-7α-O-ethyl(22*),6-O-acetyl-7β-O-ethyl(23*),cornusglucoside A(24*),comusglucoside B(25*),comusglucoside C(26*),comusglucoside D(27*),cornusglucoside E(28*),cornusglucoside F(29*),comusglucoside G(30*),cornusglucoside H(31*),williamsoside D(32),loganin(33),8-epiloganin(34),7β-O-morroniside(35),7α-O-morroniside(36),7β-O-methyl morroniside(37),7α-O-methyl morroniside(38),7β-O-ethyl morroniside(39),7α-O-ethyl morroniside(40),sweroside(41),sarracenin(42),sachalinoide B(43),(4α)-3-(5,5-dimethyltetrahydrofuranyl)-1-buten-3-ol 3-O-β-D-glucopyranoside(44),apigenin(45),chrysoderol(46),luteolin(47),quercetin(48),dihydroapigenin(49)3,4,3',4'-tetrahydroxy-δ-truxinate(50),(-)-isolariciresinol 3α-O-β-D-glucopyranoside(51),(+)-isolariciresinol 3α-O-/β-D-glucopyranoside(52),(+)-pinoresinol(53),(+)-epipinoresinol(54),syringaresinol(55),(-)-episyringaresinol(56),(-)-medioresinol(57),(7S,8R)-urolignoside(58),glochidioboside(59),(7R,8S)-dihydrodehydrodiconiferyl alcohol 9-O-β-D-glucopyranoside(60),(-)-secoisolariciresinol-9'-O-β-D-glucopyranoside(61),4-0-(6'-O-galloyl-β-D-glucopyranosyl)-cis-pcoumaric acid(62),5-(1 '-hydroxyethyl ethylenediamine)-methyl nicotinate(63),cirsiumaldehyde(64),5-hydroxymethylfurfural(65),p-Hydroxybenzoic acid(66),ethyl gallate(67),syringate(68),p-coumaric acid(69),caffeic acid(70),caffeic acid methyl ester(71),p-methoxycinnamicacid(72)。其中化合物 1～31 为新化合物,化合物42～47,49～～64和72为首次从山茱萸属植物中分离得到。基于荧光素酶报告基因技术研究化合物1～15和32～41对STAT3通路的影响,评价其抗炎活性。实验结果显示化合物3,12,33,35,36对STAT3通路表现出明显的抑制活性且未表现出细胞毒性,构效关系研究表明:a.化合物的C-7"为β构型且与葡萄糖的C-3'脱水缩合形成醚键时,对STAT3通路表现出明显的抑制活性;b.化合物的C-7"为α构型且与葡萄糖的C-6'脱水缩合形成醚键时对STAT3通路表现出明显的抑制活性;c.马钱苷和7α,β-莫诺苷对STAT3通路表现出明显的抑制活性。采用脂多糖(LPS)诱导小鼠腹腔巨噬细胞(RAW264.7)模型,采用ELISA试剂法检测化合物对细胞分泌的TNF-α、IL-6的抑制作用,评价化合物32～41的抗炎活性。实验结果表明化合物33对炎性因子TNF-α分泌的抑制作用超过50%,其余化合物未显示出抑制效果。此外,还对化合物1～15和32～41进行了 PC12细胞缺氧缺糖损伤保护作用的筛选。实验结果表明筛选的25个化合物中,化合物1和3与OGD/R相比,细胞存活率提高,说明化合物1和3对OGD/R损伤的PC12细胞物有一定的保护作用。
[Abstract]:The Cornus officinalis is the dry mature pulp of the Cornaceae plant of Cornus officinalis (Cornaceae) Cornus officinalis Sieb.et Zucc.. It mainly produced in the province of Shaanxi, Shanxi, Henan and other provinces. It has been used as the traditional medicinal herbs in China. Its medicinal history has been used for more than two thousand years. "Light body", its sexual acid, astringent, micro temperature, return to the liver, kidney meridian, with the effect of liver and kidney and astringent essence. Cornus is the main medicine of some classic Chinese medicine, such as six flavored Rehmannia pills, Jingui kidney qi pill, Zuo GUI pill and so on. The chemical composition of the fruit is mainly composed of eidolene, tannins, flavonoids, three terpenoids and phenylpropanoids. The study shows that the Fructus Corni showed good activity in the aspects of anti inflammation, prevention and control of diabetes, protection of cardiovascular and cerebrovascular, neuroprotection, antitumor, antioxidation and anti aging. Although the Fructus cornel contains a variety of pharmacological activities, its research is mostly concentrated on the level of crude extract, but few of the monomers are reported. It is of great significance to systematically study the chemical components of Cornus officinalis. This paper systematically separates and identifies the chemical constituents of the aqueous extracts of Cornus officinalis, and the preliminary evaluation of the anti-inflammatory and neuroprotective effects of the isolated monomers, for the further opening of Cornus officinalis. According to the characteristics of the maximum UV absorption of the iridoid compounds at 230 nm, 80 compounds were isolated and purified from the Fructus Corni by silica gel column chromatography, ODS column chromatography, SephadexLH-20 column chromatography and preparation high performance liquid chromatography, according to the characteristics of the maximum UV absorption of the iridoid compounds, and 80 compounds were isolated and purified from the Cornus officinalis by means of silica gel column chromatography, SephadexLH-20 column chromatography and preparation of high performance liquid chromatography. According to UV, IR, MS, HRESIMS, 1DNMR, 2DNMR, CD, ECD and other technical methods, identified the structure of 72 of these compounds, including 41 iridoid terpenoids, 3 monoterpenes, 5 flavonoids, 13 lignans and 10 other types of compounds (the compounds are named as cornuside A (1*), cornuside B (2*), cornusid, respectively. E C (3*), cornuside D (4*), cornuside E (5*), cornuside F (6*). C (20*), cornusfuroside D (21*), 6'-O-acetyl-7 alpha -O-ethyl (22*), 6-O-acetyl-7 beta -O-ethyl (23*). -epiloganin (34), 7 beta -O-morroniside (35), 7 alpha -O-morroniside (36), 7 beta -O-methyl morroniside (37), 7 alpha -O-methyl morroniside (38), 7 beta -O-ethyl morroniside (39), 7 alpha -O-ethyl morroniside (40), sweroside. (44), apigenin (45), chrysoderol (46), luteolin (47), quercetin (48), dihydroapigenin (49) 3,4,3', 4'-tetrahydroxy- Delta -truxinate (50), (-) -isolariciresinol 3 alpha -O- beta -D-glucopyranoside (51), (+) -isolariciresinol 3 alpha / beta (52), (+), (+), (54), (55), (-), (-) Garesinol (56), (-) -medioresinol (57), (7S, 8R) -urolignoside (58), glochidioboside (59), (7R, 8S) -dihydrodehydrodiconiferyl alcohol 9-O- beta -D-glucopyranoside (60), (62), 1 Mine) -methyl nicotinate (63), cirsiumaldehyde (64), 5-hydroxymethylfurfural (65), p-Hydroxybenzoic acid (66), ethyl gallate (67), Syringate (68), p-coumaric acid (69), 71 (71), 72. Compound 1~31 is a new compound, compound 42 ~ 64 and 72 For the first time isolated from the genus cornel. The effect of compound 1~15 and 32~41 on the STAT3 pathway was evaluated based on luciferase reporter gene technique and its anti-inflammatory activity was evaluated. The experimental results showed that compound 3,12,33,35,36 showed obvious inhibitory activity to the STAT3 pathway and did not show cytotoxicity. The structure-activity relationship study showed that: a. When the compound's C-7 "is a beta conformation and condenses with glucose C-3'dehydration to form an ether bond, the STAT3 pathway shows obvious inhibitory activity. The C-7" of the B. compound shows an obvious inhibitory activity to the STAT3 pathway when it is formed by condensation of the C-6' dehydration of glucose to the STAT3 pathway, and C. and 7 alpha and beta Monod glucoside are shown to the STAT3 pathway. The inhibitory activity of the mouse peritoneal macrophage (RAW264.7) was induced by lipopolysaccharide (LPS). The inhibitory effect of the compound on the secretion of TNF- A and IL-6 was detected by ELISA reagent, and the anti-inflammatory activity of compound 32~41 was evaluated. The experimental results showed that the inhibitory effect of compound 33 on the secretion of inflammatory factor TNF- alpha was more than 50%, and the remainder was found. The inhibitory effects were not shown in the compound. In addition, the protective effects of PC12 cells 1~15 and 32~41 on the protection of hypoxia and glucose deficiency were also screened. The results showed that the cell survival rate of the compounds 1 and 3 of the selected 25 compounds increased, indicating that compounds 1 and 3 had certain protective effects on the PC12 cells damaged by OGD/R.
【学位授予单位】：北京中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R284.1

【参考文献】