Enzymatic assays and molecular modeling studies of Schisandra chinensis lignans and phenolics from fruit and leaf extracts

Abstract

Considerable interest has been shown in natural sources and their compounds in developing new therapeutically agents for different diseases. In this framework, investigations performed on this topic play a central role for human health and drug development process. Schisandra chinensis (Turcz.) Baill is a medicinal and edible plant showing highly advantageous bioactivity and nutritional value. The main bioactive compounds from its fruits are lignans, derivatives of dibenzocyclooctadiene whereas concerning its leaves, phenolic acids, and flavonoids are dominant. The purpose of this study was to investigate the enzyme inhibitory potential on selected carbohydrate hydrolases, cholinesterases, and tyrosinase of extracts from fruits and leaves of Schisandra in relation with their main bioactive compounds. Furthermore, the interactions between dominant compounds (schisandrol A, schisandrol B, schisandrin B, and cinnamic acid) from extracts and selected enzymes were investigated by molecular modeling and molecular dynamic studies in order to explain at a molecular level our findings.     

Characteristics and Antioxidant Activity of Lignans in Schisandra chinensis and Schisandra sphenanthera from Different Locations

Twenty Schisandra samples were collected from different locations. Contents of 7 lignans in the samples were determined and analyzed by HPLC method coupled with hierarchical clustering analysis (HCA) and principal component analysis (PCA), and the antioxidant capacity of Schisandra from the different locations was evaluated by reducing power, ferric thiocyanate (FTC) and 2,2′‐diphenyl‐1‐picrylhydrazyl (DPPH) assay. The results showed that there was a significant difference in the content of lignans between Schisandra chinensis and Schisandra sphenanthera. The Schisandra sphenanthera samples in the southwest of China were significantly different from those from the other locations. The antioxidant capacity of Schisandra chinensis was significantly superior to that of Schisandra sphenanthera, and the main antioxidant components were schisandrol A, schisandrol B and schisandrin B based on the result of discrimination analyses. The differences in the chemical composition and antioxidant activity of lignans in Schisandra chinensis and Schisandra sphenanthera from the different locations were investigated in this study, which may provide an experimental basis for the quality control of Schisandra.     

New Thymoquinol Glycosides and Neuroprotective Dibenzocyclooctane Lignans from the Rattan Stems of Schisandra chinensis

Three new lignans ( 1 – 3 ), together with four new thymoquinol glycosides ( 4 – 7 ), were isolated from 70%‐EtOH extract of the rattan stems of Schisandra chinensis. The structures of 1 – 7 were elucidated by detailed spectroscopic analyses, and these new compounds were identified as pinobatol‐9‐O‐β‐d ‐glucopyranoside ( 1 ), 1,2,13,14‐tetramethoxydibenzocyclooctadiene 3,12‐O‐β‐d ‐diglucopyranoside ( 2 ), 3,7‐dihydroxy‐1,2,13,14‐tetramethoxydibenzocyclooctadiene 12‐O‐β‐d ‐glucopyranoside ( 3 ), thymoquinol 2‐O‐β‐d ‐apiofuranosyl‐(1→6)‐β‐d ‐glucopyranoside ( 4 ), thymoquinol 2‐O‐α‐d ‐arabinofuranosyl‐(1→6)‐β‐d ‐glucopyranoside ( 5 ), thymoquinol 5‐O‐β‐d ‐apiofuranosyl‐(1→6)‐β‐d ‐glucopyranoside ( 6 ), and thymoquinol 5‐O‐α‐d ‐arabinofuranosyl‐(1→6)‐β‐d ‐glucopyranoside ( 7 ). The neuroprotective activity of 1 – 7 was evaluated on PC12 cells with neurotoxicity induced by amyloid‐beta 1 – 42 (Aβ_{1 – 42}). Compounds 2 and 3 showed protecting activity against Aβ‐induced toxicity in PC12 cells.     

北五味子果实超临界CO2萃取工艺的研究

通过正交设计的试验方法,探讨了超临界CO₂法萃取五味子果实中木脂素的工艺条件,确定了最佳工艺条件为萃取压力30 Mpa,萃取温度50℃,萃取时间120 min。     

高效液相色谱法测定五味子乙素含量

采用高效液相色谱法,测定了五味子果实中五味子乙素的含量,检测波长为254 nm,标准曲线线性关系R²为0.9907,相对标准偏差为0.48%,平均加样回收率为99.14%。结果表明,此方法准确度较高,为五味子生药及成品药中五味子乙素含量测定提供了一种切实可行的方法。     

Terpenoids from Salvia palaestina

Six known terpenoids: vergatic acid, ursolic acid, crataegolic acid, lupane-3β-,11α, 20-triol, sclareol and sitosteryl 3β-glucoside were isolated from the leaves of Salvia palaestina and were identified by spectral data. Among the compounds, sclareol showed high activity against Staphylococcus aureus, S. epidermis, Escherichia coli, Proteus vulgaris and Pseudomonas aeruginosa, while the triterpenoids were not tested due to solubility problems.     

Terpenoids with Cytotoxic Activity from Roots of Ardisia crispa

Three new terpenoids, ardisiacrispins G-I ( 1 , 4 and 8 ), and eight known compounds, cyclamiretin A ( 2 ), psychotrianoside G ( 3 ), 3-hydroxy-β-damascone ( 5 ), megastigmane ( 6 ), corchoionol C ( 7 ), zingiberoside B ( 9 ), angelicoidenol ( 10 ), trans-linalool-3,6-oxide-β-D-glucopyranoside ( 11 ) were isolated from the roots of Ardisia crispa. The chemical structures of all isolated compounds were elucidated by extensive spectroscopic analyses, such as HR-ESI-MS, 1D and 2D NMR spectra. Ardisiacrispin G ( 1 ) represents the oleanolic-type scaffold featuring a rare 15,16- epoxy system. All compounds were evaluated for the cytotoxicity against two cancer cell lines (U87 MG and HepG2) in vitro. Compounds 1 , 8 and 9 exhibited moderate cytotoxic activity with IC₅₀ values ranging from 7.6±1.1 to 28.8±3.2 μM.     

广东紫珠中的萜类化合物及其抗炎活性

对岭南药材广东紫珠(Callicarpa kwangtungensis)地上部分进行化学成分研究,得到11个萜类化合物,分别鉴定为sambucunlin A(1)、2α-羟基羽扇豆醇(2)、swinhoeic acid(3)、3β-羟基-乌苏烷-11-烯-13β,28-内酯(4)、蔷薇酸(5)、2α,3β,6β,18β,23-pentahydroxy-olean-12-en-28-oic acid(6)、rel-5-(3S,8S-dihydroxy-1R,5S-dimethyl-7-oxa-6-oxobicyclo-oct-8-yl)-3-methyl-2Z,4E-pentadienoic acid(7)、salvionoside B(8)、齐墩果酸(9)、白桦脂酸(10)和α-香树脂醇(11)。其中,化合物1～4和6～8为首次从该属植物中分离得到。在化学成分分离基础上,进一步选择脂多糖(LPS)诱导的RAW 264.7小鼠巨噬细胞炎症模型进行萜类化合物抗炎活性测试。结果表明:化合物3和9具有显著的抗炎活性,对比结构发现,三萜类化合物(3～6、9和11)抗炎活性优于倍半萜类化合物(7和...     

A Cytotoxic Neolignan from Schisandra propinqua （Wall.） Baill.

In the course of our study of bioactive natural products from Schisandra plants, we isolated a neolignan from an EtOAc extract of the stems of Schisandra propinqua （Wall.） Baill. The structure of the new com- pound was determined to be 4, 4-di （4-hydroxy-3-methoxyphenly）-2, 3-dimethylbutanol （compound 1） on the basis of 1H- and 13C-NMR spectra and 2D NMR methods. Eight known compounds, compounds 2-9, were also isolated and identified, of which compounds 3, 4, 6 and 9 were isolated for the first time from this plant. In addition, compounds 1-4 were evaluated for cytotoxicity by an 3-（4,5-dimethyl-2 thiazoyl）-2,5-diphenyl-2H- tetrazolium bromide （MTT） assay. Compound 1 showed significant potential cytotoxic ability in the bioassay.     

Analysis of volatile fractions of Schisandra chinensis (Turcz.) Baill. using GC-MS and chemometric resolution

The two-dimensional data obtained from GC-MS has been used qualitatively and quantitatively to determine the components of the volatile fractions of Schisandra chinensis obtained by six different extraction methods. Sub-window factor analysis (SFA) was employed to confirm the identities of components determined in different samples. With the help of SFA, and other chemometric techniques, peak purity in the chromatograms was determined, and overlapping peaks were resolved to yield a pure chromatographic profile and mass spectrum for each component. It is demonstrated that the accuracy of qualitative and quantitative analysis may be greatly enhanced using chemometric resolution methods, such methods being particularly valuable with respect to the analysis of complex samples such as traditional Chinese medicines. It is further demonstrated that different extraction methods give rise to volatile fractions of S. chinensis which differ qualitatively and quantitatively in their composition.     

高压脉冲电场法快速提取北五味子中五味子醇甲的工艺研究

本文采用正交实验优化提取方法,HPLC法测定五味子醇甲含量,对五味子的快速提取工艺方法进行了研究。结果表明:以80%乙醇作为提取液,电场强度10 kv/cm,脉冲数为10,料液比1∶10为最佳提取工艺;影响因素的大小依次为:电场强度、料液比、脉冲数。此法提取五味子中的五味子醇甲,提取率可达0.76%,得膏率达27.886%。本提取工艺可行,经济、省时、回收率高。     

A compound isolated from Schisandra chinensis induces apoptosis

Schizandra chinensis has been known to have five predominant tastes: salty, sweet, sour, astringent, and bitter. It has also been shown to have various effects on the cardiovascular system, gastrointestinal system, anti-inflammatory, central nervous system, endocrine system, and stress protect. However, its anti-cancer activity on colon carcinoma HCT-116 cells has not been yet been examined. Thus, in this study, we attempted to isolate a compound from Schisandra chinensis that induced apoptosis in HCT-116 cells. An active compound was found and identified to be Gomisin A. It displayed apoptotic activity through caspase-7 cleavage in colon carcinoma HCT-116 cells. In addition, we further assessed the effects of this compound using long-term survival clonogenic assay with HCT116 cells.     

中国红豆杉枝叶中的紫杉烷二萜

从中国红豆杉(Taxus chinensis)枝叶的乙醇提取物中分离得到8个紫杉烷二萜,通过波谱分析分别确定为：14p羟基巴卡亭Ⅵ(1),巴卡亭Ⅵ(2),巴卡亭Ⅳ(3),1β-去羟基巴卡亭Ⅳ(4),云南红豆杉酯甲(5),2α-去乙酰-2α-苯甲酰基-13α-乙酰基云南紫杉亭(6),5α-羟基-2α,7βp,9α,10β,13α-五乙酰氧基紫杉-4(20),11-二烯(7)和Taxaein(8)。其中化合物1为新化合物,并报道八个化合物在丙酮中测得的核磁共振信号,化合物8的数据属首次报道。     

蛇含委陵菜的木脂素类成分及其细胞毒活性研究

为了研究蛇含委陵菜(Potentilla kleiniana)的化学成分及其肿瘤细胞毒活性,该研究综合运用D-101大孔树脂、硅胶、Sephadex LH-20、Toyopearl HW-40F及半制备高效液相等现代色谱分离技术对蛇含委陵菜60%乙醇提取物进行分离纯化,根据化合物的理化性质结合核磁共振波谱(NMR)、高分辨质谱(HR-ESI-MS)鉴定化合物的结构,并采用MTT法测定各化合物对人宫颈癌细胞株Hela的细胞毒活性。结果表明:(1)从蛇含委陵菜中分离鉴定了13个木脂素类化合物,分别为(+)-松脂素(1)、(+)-8-羟基松脂素(2)、(+)-丁香脂素(3)、(+)-杜仲树脂酚(4)、(+)-松脂素-4-O-β-D-吡喃葡萄糖苷(5)、(+)-8''-羟基松脂素-4-O-β-D-吡喃葡萄糖苷(6)、(+)-8''-羟基松脂素-4''-O-β-D-吡喃葡萄糖苷(7)、(+)-松脂素-8''-O-β-D-吡喃葡萄糖苷(8)、schilignan F(9)、(+)-松脂素-4, 4''-O-双吡喃葡萄糖苷(10)、(+)-落叶松脂素-4''-O-β-D-吡喃葡萄糖苷(11)、neoolivil-4-O-β-D-glucopyranoside(12)、3,3''-bis [3,4-dihydro-4-hydroxy-6-methoxy-2H-1-benzopyran](13)。其中,化合物1-4、7、8、10、12、13为首次从委陵菜属植物中分离得到,化合物5、6、9、11为首次从蛇含委陵菜中分离得到。(2)细胞毒试验结果显示,化合物1、3、4对Hela细胞具有较好的抑制活性,其半数抑制浓度IC₅₀值分别为(69.94±1.89)、(66.25±2.11)、(59.81±1.73)μmol·L^-1。该研究结果进一步丰富了蛇含委陵菜的化学成分,为抗宫颈癌药物的研发提供物质基础。     

Influence of mannan oligosaccharide,Ligustrum lucidum and Schisandra chinensis on parameters of antioxidative and immunological status of broilers

Abstract

The study was conducted to evaluate effects of dietary supplementation with Ligustrum lucidum (LL, 10 g/kg), Schisandra chinensis (SC, 10 g/kg), LL (10 g/kg) + mannan oligosaccharides (MOS, 50 mg/kg), or SC (10 g/kg) + MOS (50 mg/kg) on growth performance and parameters of antioxidative and immunological status of broilers. The results showed that feeding LL, SC, LL + MOS, or SC + MOS had no significant effect on growth performance of broilers relative to the control. However, compared to the control, LL, SC, LL + MOS, or SC + MOS significantly decreased malondialdehyde concentration in serum, thigh, and heart of broilers. In addition, glutathione reductase activity of heart and sera of the birds were significantly elevated by supplementation LL, SC, LL + MOS, or SC + MOS. Furthermore, LL, SC, LL + MOS, or SC + MOS significantly improved antibody titres against Newcastle disease virus and lymphocyte proliferation of broilers (p < 0.05). Whereas, no cooperating effect between LL (or SC) and MOS on antioxidant status and immunity of broilers were found.     

Dimethylbutyrylated Dibenzocyclooctadiene Lignans from the Fruits of Schisandra cauliflora Inhibit NO Production in LPS-Activated RAW264.7 Cells

From the fruits of Schisandra cauliflora, five new dimethylbutyrylated dibenzocyclooctadiene lignans, named schisandracaurins A−E, were isolated using separation and chromatographic techniques. Their structures were determined by extensive analyses of HR-ESI-MS, NMR, and ECD spectra. The schisandracaurins A−E potentially inhibited NO production in LPS-activated RAW264.7 cells with their IC₅₀ values from 21.4 to 30.3 μM.     

胚乳和外源赤霉素对五味子[Schisandra chinensis(Turcz.)Baill]种子发芽的影响

以组织培养和土壤播种的未经层积处理的五味子种子为材料,检测胚乳和外源赤霉素(GA3)对发芽影响的结果表明,在不加生长调节物质的MS培养基上,去掉外种皮带全胚乳的种子发芽率只有2.7%,而带1/2和1/3胚乳的种子发芽率则分别为46%和70.6%;培养基中添加GA3,可提高去掉外种皮带全胚乳和带1/2胚乳种子的发芽率,而对带1/3胚乳的种子则无效果.土壤播种时,用10～1000mg·L-1GA3对去掉外种皮种子进行1～10d前处理,其中100mg·L-1GA3前处理10d的发芽率最高,达到38.7%.