基于基因芯片技术的五味子素B诱导高甘油三酯血症小鼠模型作用机制研究＊

目的 探讨五味子素B（Schisandrin B，Sch B）诱导高脂血症小鼠模型的基因（肝脏）表达谱的变化及可能的机制。方法 ICR小鼠，雄性，分为7组，每组10只：正常饮食（normal diet，ND）组（2组）、ND/Sch B（0.5，2 g·kg^-1）组、高脂/糖饮食（high fat/fructose diet，HFFD）组、HFFD/Sch B（0.5，2 g·kg^-1）组。小鼠用Sch B（橄榄油配制）灌胃，48 h后用生化法检测血清和肝脏甘油三酯（TG）和总胆固醇（TC）水平，血清高密度脂蛋白胆固醇（HDL）和低密度脂蛋白胆固醇（LDL）水平；ELISA法检测血清极低密度脂蛋白胆固醇（VLDL）、载脂蛋白A1（Apo A1）、Apo E、Apo B48、Apo B100、Apo CⅡ和Apo CⅢ水平；HE染色观察肝脏病理学变化；油红O染色观察肝脏脂质沉积；通过基因芯片技术分析各组小鼠肝脏中全基因表达谱的变化及所涉及的通路。结果 ND/Sch B组小鼠血清/肝脏TG（升高266%/352.57%）和TC水平显著升高（升高36.54%/25.70%），血清HDL和LDL水平显著升高（分别升高29.96%和30.68%），但VLDL、Apo E、Apo B100、Apo CⅡ和Apo CⅢ显著降低（分别降低15.98%、28.90%、20.76%、20.53%和17.82%）；肝脏脂质沉积并出现病理学损伤；与HFFD组比，HFFD/Sch B组血清TG水平、肝脏TG和TC水平升高，血清HDL、LDL、VLDL、Apo E、Apo B100和Apo CⅡ水平显著降低；ND/Sch B组与ND组比肝脏有1016个基因差异表达，其中上调基因722个，下调基因294个，HFFD/Sch B组与HFFD组比肝脏有1162个基因差异表达，其中上调基因671个，下调基因491个。而HFFD组与ND组比肝脏有2070个基因差异表达，其中上调基因1289个，下调基因781个。Pathway分析结果显示Sch B诱导的高脂血症与11条通路的改变有关。结论 Sch B诱导高脂血症的机制与肝脏调节脂代谢多条通路、载脂蛋白及脂蛋白代谢密切相关。     

五味子乙素防护晶状体氧化损伤的体外实验研究

 目的探讨五味子乙素(Sch B)对实验性晶状体氧化损伤的防护作用。方法采用Fenton反应复制兔晶状体氧化损伤模型,以吡诺克辛钠滴眼液(PS)为对照,测定晶状体SP,SOD,GSH-Px,GSH,VitC及MDA的含量,观察终浓度0.5 mmol·L^-1Sch B对晶状体氧化损伤的防护作用。结果Sch B组与Fenton对照组比较,Sch B可显著减少Fenton反应中SP的丢失,显著提高SOD和GSH-Px的活性及GSH和Vit C的水平,并降低MDA含量。Sch B组与PS组比较,Sch B组的SOD活性、GSH含量和Vit C水平分别是PS组的1.66倍,2.58倍和2.36倍,MDA含量比PS组下降24%。上述各项指标均有显著性差异(P＜0.01)。结论 Sch B对Fenton反应中晶状体的氧化损伤有明显的防护作用,且优于吡诺克辛钠滴眼液。     

Schisandrin B attenuates epidural fibrosis in postlaminectomy rats by inhibiting proliferation and extracellular matrix production of fibroblasts

Laminectomy has been widely considered one of the most common treatments for lumbar disorders. Epidural fibrosis (EF) is a common complication after laminectomy, causing recurrent postoperative pain. Schisandrin B (Sch.B), the active ingredient extracted from Schisandra chinensis Fructus, has been found to have potent antiproliferative and antifibrotic effects on several cells. This study aimed to investigate the effects of Sch.B on the prevention of postlaminectomy EF formation. In vitro, we studied the effects of Sch.B on transforming growth factor beta 1 (TGF‐β1)‐induced proliferation and extracellular matrix (ECM) production of primary fibroblasts, as well as its underlying mechanism. We found that Sch.B not only inhibited the proliferation of fibroblasts but also reduced ECM production, including that of connective tissue growth factor, fibronectin, and type I collagen, in a dose‐dependent manner. Mechanistically, we found that Sch.B suppressed TGF‐β1‐stimulated activation of the Smad2/3 and mitogen‐activated protein kinase pathways. Moreover, the in vivo study demonstrated that Sch.B treatment attenuated the progression of EF in a postlaminectomy rat model via reducing the cell number and ECM production of scar tissue. Taken together, these data suggested that Sch.B possesses great potential value as a preventative agent for EF.     

Chemokine receptor CCR-5 inhibitors produced by Chaetomium globosum

Two novel chemokine receptor CCR-5 inhibitors, Sch 210971 (1) and Sch 210972 (2), were isolated from the fungal fermentation broth of Chaetomium globosum by normal- and reversed-phase HPLC purifications. The structure determination of 1 and 2 was accomplished on the basis of UV, MS, and NMR spectral data analyses including COSY, NOESY, HMQC, and HMBC experiments. The structure and relative configuration of 2 were determined unequivocally by X-ray crystallographic analysis. The major component 2 demonstrated a potent inhibitory activity of IC(50) = 79 nM in the CCR-5 receptor in vitro binding assay.     

Schisandrin C targets Keap1 and attenuates oxidative stress by activating Nrf2 pathway in Ang II‐challenged vascular endothelium

Vascular endothelial dysfunction plays a crucial role in the pathogenesis of cardiovascular diseases. Oxidative stress is a key pathophysiological mechanism underpinning endothelial dysfunction. Schisandrin C (Sch C), a dibenzocyclooctadiene derivative of Schisandra chinensis, has antioxidative properties. Here, we report the use of Sch C as a novel therapeutic for the treatment of angiotensin II (Ang II)‐induced endothelial deficits and explore the underlying mechanisms and the target of Sch C. Our results demonstrated that Sch C treatment prevents aorta oxidative stress and improves relaxation in mice, challenged with subcutaneous infusion of Ang II. In addition, Sch C significantly ameliorates Ang II‐induced oxidative stress in rat aortic endothelial cells. We then discovered that these antioxidative effects of Sch C are mediated through the induction of nuclear factor (erythroid‐derived 2)‐like 2 (Nrf2). Using an expression plasmid and molecular docking, we identified that Kelch‐like ECH‐associated protein‐1 (Keap1), a negative regulator of Nrf2, is a target of Sch C. These findings provide evidence for the potential use of Sch C as an antioxidative agent for treatment of vascular endothelial deficits.     

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??OBJECTIVE To investigate the synergetic effect of schisandrin B and liposomes on overcoming multidrug resistance(MDR). METHODS Co-delivery of doxorubicin and schisandrin B by liposome(DS-L) were prepared, and doxorubicin solution(F-DOX), schisandrin B and doxorubicin mixture(Sch B+DOX), doxorubicin liposomes(D-L), doxorubicin liposomes and schisandrin B mixture(D-L+Sch B), verapamil and doxorubicin mixture(Ver+DOX) were also prepared as the control. The MTT test were measured, and the amount of doxorubicin in the K562/DOX cells at different time were determined, and time course of uptake and efflux were drawn. RESULTS The MTT test shows that the resistance factor(RF) of group DS-L were 1.68, 14.52 and 1.42 times of group Sch B+DOX, group D-L and group D-L+Sch B respectively. The uptake test shows that amount of doxorubicin of K562/DOX cells in group DS-L was 1.30 and 1.21 times of that in group D-L and group Sch B+DOX respectively. And the efflux test shows DS-L could delay doxorubicin efflux from K562/DOX cells. CONCLUSION The co-delivery of chemotherapeutics and P-gp inhibitors schisandrin B by liposome is a promising approach to overcome MDR. And there is a synergistic effect between liposome and schisandrin B to overcome MDR.     

Nematicidal epipolysulfanyldioxopiperazines from Gliocladium roseum

Five new verticillin-type epipolysulfanyldioxopiperazines, gliocladine A (1), B (2), C (3), D (4), and E (5), were isolated from wheat solid-substrate fermentation of Gliocladium roseum 1A, along with four known compounds, verticillin A (6), 11'-deoxyverticillin A (7), Sch52900 (8), and Sch52901 (9). Their structures were elucidated by extensive 1D and 2D NMR studies, MS, and chemical transformations. In vitro immersion tests showed that all nine compounds exhibited antinematodal activity against Caenorhabditis elegans and Panagrellus redivivus. The monomeric epipolysulfanydioxopiperazines (3-5), with the indole moiety, were found to be less active than the dimeric compounds (1, 2, 6-8).     

Protective effect of schisandrin B against cyclosporine A-induced nephrotoxicity in vitro and in vivo

Schisandrin B (Sch B) is an active ingredient of the fruit of Schisandra chinensis. It has many therapeutic effects arising from its tonic, sedative, antitussive and antiaging activities and is also used in the treatment of viral and chemical hepatitis. The aim of this study was to investigate the protective effects of Sch B on cyclosporine A (CsA)-induced nephrotoxicity in mice and HK-2 cells (a human proximal tubular epithelial cell line). After gavage with Sch B (20 mg/kg) or olive oil (vehicle), mice received CsA (30 mg/kg) by subcutaneous injection once daily for four weeks. Renal function, histopathology, and tissue glutathione (GSH) and malondialdehyde (MDA) levels were evaluated after the last treatment. The effects of Sch B on CsA-induced oxidative damage in HK-2 cells were investigated by measuring cell viability, the release of lactate dehydrogenase (LDH), the level of reactive oxygen species (ROS), and the cellular GSH and ATP concentrations. Cellular apoptosis was assessed by flow cytometry. Treatment with Sch B in CsA-treated mice significantly suppressed the elevation of blood urea nitrogen (BUN) and serum creatinine levels and attenuated the histopathological changes. Additionally, Sch B also decreased renal MDA levels and increased GSH levels in CsA-treated mice. Using an in vitro model, Sch B (2.5, 5 and 10 μM) significantly increased the cell viability and reduced LDH release and apoptosis induced by CsA (10 μM) in HK-2 cells. Furthermore, Sch B increased the intracellular GSH and ATP levels and attenuated CsA-induced ROS generation. In conclusion, Sch B appears to protect against CsA-induced nephrotoxicity by decreasing oxidative stress and cell death.     

Schisandrin B stereoisomers protect against hypoxia/reoxygenation‐induced apoptosis and associated changes in the Ca2+‐induced mitochondrial permeability transition and mitochondrial membrane potential in AML12 hepatocytes

The effects of the schisandrin B stereoisomers, (±)γ‐schisandrin [(±)γ‐Sch] and (?)schisandrin B [(?)Sch B], on hypoxia/reoxygenation‐induced apoptosis were investigated in AML12 hepatocytes. Changes in cellular reduced glutathione (GSH) levels, Ca²⁺‐induced mitochondrial permeability transitions (MPTs) and mitochondrial membrane potentials (Δψ_m values) were also examined in (±)γ‐Sch‐ and (?)Sch B‐treated cells, without or with hypoxia/reoxygenation challenge. The (±)γ‐Sch/(?)Sch B pretreatments (2.5–5.0 µm ) protected against hypoxia/reoxygenation‐induced apoptosis in AML12 cells in a concentration‐dependent manner, with the (?)Sch B effect being more potent. Drug antiapoptotic effects were further evidenced by suppression of hypoxia/reoxygenation‐induced mitochondrial cytochrome c release and subsequent cleavage of caspase 3 and poly‐ADP‐ribose polymerase by (?)Sch B pretreatment. Whereas hypoxia/reoxygenation challenge increased the extent of Ca²⁺‐induced MPT pore opening, and Δψ_m, in AML12 hepatocytes, cytoprotection afforded by (±)γ‐Sch/(?)Sch B pretreatment against hypoxia/reoxygenation‐induced apoptosis was associated with a decreased sensitivity to Ca²⁺‐induced MPT and an increased Δψ_m in both unchallenged and challenged cells, compared with the drug‐free control. The results indicate that (±)γ‐Sch/(?)Sch B pretreatment protected against hypoxia/reoxygenation‐induced apoptosis in AML12 hepatocytes and that the cytoprotection afforded by (±)γ‐Sch/(?)Sch B may at least in part be mediated by a decrease in sensitivity to Ca²⁺‐induced MPT, which may in turn result from enhancement of cellular GSH levels by drug pretreatments. Copyright © 2009 John Wiley & Sons, Ltd.     

天然抗氧化剂对抗PC12细胞氧化损伤的实验研究

目的:研究五味子乙素(SchB)对H2O2诱导的PC12细胞氧化损伤的保护作用及作用机制。方法:SchB与PC12细胞预培养24h后,加入H2O2处理,在倒置显微镜下观察细胞的形态改变,采用MTT法测定细胞生存率,采用单细胞凝胶电泳法测定DNA单链损伤,采用荧光法测定细胞内的谷胱甘肽(GSH)含量。结果:H2O2处理后细胞生存率明显降低,细胞内DNA单链损伤明显升高,而GSH含量明显降低。经SchB预培养的细胞生存率明显升高,细胞内DNA单链损伤明显减轻,GSH含量明显升高,其作用呈剂量依赖关系。结论:SchB对PC12细胞有明显的保护作用,其作用机制可能与提高GSH抗氧化系统的能力有关。     

民族药马兰的生药学鉴定

目的对民族药马兰进行系统的生药学鉴定,为其鉴别提供科学依据。方法采用原植物、性状、显微、薄层鉴定方法。结果通过原植物、性状鉴别、显微鉴别、薄层色谱鉴别能够很好地鉴定原植物。结论这些方法简便易操作,可作为该民族药材定性鉴别方法。     

Isolation and structure elucidation of Sch 642305, a novel bacterial DNA primase inhibitor produced by Penicillium verrucosum

A novel primase inhibitor, Sch 642305 (1), was isolated from the fermentation broth of the fungal culture Penicillium verrucosum. The structure of 1 was elucidated on the basis of MS and NMR spectroscopic data as a new and unusual bicyclic 10-membered macrolide. The absolute configuration of the asymmetric centers was determined by X-ray crystallographic analysis of the p-bromobenzoate derivative (3). Compound 1 exhibited inhibitory activity against bacterial DNA primase enzyme with an EC(50) of 70 microM.     

Pharmacological aspects of selected herbs employed in Hispanic folk medicine in the San Luis Valley of Colorado, USA: I. Ligusticum porteri (osha) and Matricaria chamomilla (manzanilla)

Interviews with Hispanic families in the San Luis Valley of Colorado delineated several medicinal herbs that are popular in Hispanic folk medicine, including Ligusticum porteri (osha) and Matricaria chamomilla (manzanilla). A search of the scientific literature reveals that related species of Ligusticum and Matricaria chamomilla contain compounds that possess significant pharmacologic activity. This combined information is now being used as a basis for further investigation at the University of Colorado School of Pharmacy in an effort to detect pharmacologic activity in osha and manzanilla preparations.     

细胞凋亡抑制剂防护晶状体上皮细胞氧化损伤及信号转导机制

目的：探讨细胞凋亡抑制剂阿魏酸钠（SF）、五味子乙素（Sch B）、菊花（FC）、车前子（SP）对过氧化氢（H2O2）所致氧化损伤的晶状体上皮细胞（LEC）活性以及细胞内钙（Ca2＋）、环磷酸腺苷（cAMP）、环磷酸鸟苷（cGMP）的影响,研究四种细胞凋亡抑制剂防治白内障的细胞信号转导机制。方法：将传代培养的牛LEC与H2O2共同孵育复制氧化损伤的模型,同时加入SF和Sch B单体及FC和SP水提取液孵育后,采用四甲基偶氮唑蓝法（MTT）检测LEC活性,采用荧光分光光度法检测LEC内钙离子的浓度（[Ca2＋]i）、放射免疫分析法检测LEC内cAMP和cGMP含量,并探讨不同浓度和不同孵育时间（12h、24h、36h）的影响。结果：①H2O2组LEC活性下降,SF、Sch B、FC、SP均可明显增强氧化损伤的LEC活性（P〈0.01）,呈浓度依赖关系和时间依赖关系。②H2O2组LEC的[Ca2＋]i升高,SF、Sch B、FC、SP均可显著降低氧化损伤LEC的[Ca2＋]i（P〈0.01）,呈时间-效应关系。③H2O2组LEC的cAMP浓度升高、cGMP浓度下降,SF、Sch B、FC、SP均可使氧化损伤的LEC的cAMP水平显著降低（P〈0.01）、cGMP水平显著升高（P〈0.01）,也呈时间-效应关系。结论：四种细胞凋亡抑制剂SF、Sch B、FC、SP可明显抑制LEC氧化损伤及凋亡,通过[Ca2＋]i、cAMP、cGMP信号转导及相互作用调节生物学效应可能是其主要的细胞和分子生物学机制。     

Pinophilins A and B, inhibitors of mammalian A-, B-, and Y-family DNA polymerases and human cancer cell proliferation

Pinophilins A (1) and B (2), new hydrogenated azaphilones, and Sch 725680 (3) were isolated from cultures of a fungus (Penicillium pinophilum Hedgcok) derived from a seaweed, and their structures were determined using spectroscopic analyses. These compounds selectively inhibited the activities of mammalian DNA polymerases (pols), A (pol γ), B (pols α, δ, and ε), and Y (pols η, ι, and κ) families, but did not influence the activities of the four X-family pols (pols β, λ, μ, and terminal deoxynucleotidyl transferase). Compound 1 was the strongest inhibitor, with IC?? values of 48.6 to 55.6 μM. Kinetic analysis showed that compound 1 is a noncompetitive inhibitor of both pol α and κ activities with the DNA template-primer substrate, and a competitive inhibitor with the nucleotide substrate. In contrast, compounds 1-3 showed no effect on the activities of plant and prokaryotic pols or any other DNA metabolic enzymes tested. The compounds suppressed cell proliferation and growth in five human cancer cell lines, but had no effect on the viability of normal human cell lines.     

Plants used for the treatment of diabetes in Israel

In an extensive ethnobotanical survey (130 informants) of the medicinal plants of Israel, 16 species were found to be used for hypoglycaemic treatments. The list includes Achillea fragrantissima (Forssk.) Sch.-Bip, Ammi visnaga (L.) Lam, Atriplex halimus L., Capparis spinosa L., Ceratonia siliqua L., Cleome droserifolia (Forssk.) Del., Eryngium creticum Lam., Inula viscosa (L.) Ait., Matricaria aurea (Loefl.) Sch.-Bip, Origanum syriaca L., Paronychia argentea Lam, Prosopis farcta (Banks et Sol.) Macbride, Salvia fruticosa Mill., Sarcopoterium spinosum (L.) Sp., and Teucrium polium L.; eight of them (marked with an asterisk) are first recorded here as used for this purpose.     

Isolation, partial characterisation and immunomodulating activities of polysaccharides from Vernonia kotschyana Sch. Bip. ex Walp

The roots from Vernonia kotschyana Sch. Bip. ex Walp. (Baccharoides adoensis var. kotschyana (Sch. Bip. ex Walp.) M.A. Isawumi, G.El-Ghazaly & B. Nordenstam) (Asteraceae) are used in Malian folk medicine for the treatment of gastritis, gastro duodenal ulcers, as an aid to ameliorate digestion and as a wound healing remedy. Since a common feature among these conditions is related to immune responses, immunomodulating activities of fractions isolated from both the 50 degrees C and the 100 degrees C water extracts from Vernonia kotschyana were investigated in this study. The active principles were identified as acidic polysaccharide fractions, containing pectic arabinogalactan type II structures, which showed both complement fixing ability and T-cell independent induction of B-cell proliferation in vitro. Some activity was also observed on macrophages. The present study may provide additional support for the popular use of this plant to improve intestinal health.     

补肾中药对肾阳虚大鼠下丘脑—垂体—肾上腺轴Fos蛋白表达的影响

目的：研究c-Fos蛋白在肾阳虚发病中的作用及补肾中药的调整作用。方法：应用免疫染色技术检测肾阳虚大鼠下丘脑－垂体－肾上腺轴c-Fos蛋白的表达。结果：肾阳虚大鼠下丘脑视交叉上核和肾上腺组织中c-Fos阳性细胞数显著升高,补肾中药能够抑制肾阳虚大鼠下丘脑我叉上核和肾上腺组织c-Fos的升高。结论：下丘脑视交叉上核和肾上腺c-Fos蛋白的过度表达可能是肾阳虚证的病机之一,补肾中药可通过抑制下丘脑视交叉上核和肾上腺组织中c-Fos的表达,而改善下丘脑－垂体－肾上腺轴的功能,起到治疗作用。     

清热活血法为主治疗小儿紫癜性肾炎14例

应用清热解毒、活血化瘀法为主治疗14例紫癜性肾炎,其中轻型2例,肾炎型10例,肾炎肾病型2例。中医辨证多为血热型或血热夹瘀型。结果13例痊愈,1例好转;见效时间平均1.7月;平均随访1.58年未见复发。认为用清热活血法疗效满意。对轻型或肾炎型紫癜性肾炎单用中药治疗为好。     

Anti-hypoxic activity at simulated high altitude was isolated in petroleum ether extract of Saussurea involucrata

Ethnopharmacological relevance

Rhodiola algida, Saussurea involucrata, and other herbs grown in Qinghai-Tibetan plateau have long been used to prevent and treat acute mountain sickness.

Aim of the study

To screen and identify the anti-hypoxic constituents in the herbs grown in Qinghai-Tibetan plateau of Northwestern China.

Materials and methods

The anti-hypoxic activities of 20 selected plateau herbs were examined against two positive controls, Rhodiola algida and acetazolamide, using the normobaric hypoxia model of mice. The herb with the highest activity was successively extracted with 70% ethanol, petroleum ether, chloroform, ethyl acetate and n-butanol. The extract with the highest activity was identified by comparing the survival time of mice under normobaric hypoxia condition after being subjected to different extracts. The identified extract was further tested by simulating high altitudes through an acute decompression model and a chronic decompression model for mice.

Results

The herb found to have the highest anti-hypoxic activity was Saussurea involucrate (Kar. et Kir.) Sch.-Bip, and the most effective fraction was in the petroleum ether extract. Administration of petroleum ether extract of Saussurea involucrata (PESI) to mice at 50 mg/kg significantly decreased the mortality of animals under acute decompression conditions. Changes in biochemical indicators for glycometabolism and energy metabolism, including adenosine triphosphate (ATP) content and adenosine triphosphatase (ATPase) activity in brain and cardiac muscle, lactic acid (LAC) and lactate dehydrogenase (LDH) in blood and cardiac muscles, blood sugar, and glycogen content in liver and skeletal muscle were reversed under chronic decompression conditions.

Conclusions

Saussurea involucrata (Kar. et Kir.) Sch.-Bip exhibits high anti-hypoxic activity that may be effective in preventing acute mountain sickness, and the active constituents are mainly in the petroleum ether extract.