Cytotoxicity and anti-hepatitis B virus activities of saikosaponins from Bupleurum species

Saikosaponins, the main active constituents of Bupleurum spp., have been shown to possess immunomodulatory, hepatoprotective, anti-tumor and anti-viral activities. In this study, saikosaponins a, c and d were evaluated for cytotoxicity and anti-hepatitis B virus ( HBV) activities. Results showed that, with the exception of saikosaponins a and d, HBV-transfected human hepatoma cells (2.2.15 cells) cultured with saikosaponin c showed a significantly lower level of HBeAg in culture medium. Saikosaponin c also possessed activity in inhibiting HBV DNA replication; this inhibitory effect was not due to the cytotoxicity of saikosaponin c or its effect on 2.2.15 cell proliferation. Although saikosaponin d exhibited cytotoxicity on 2.2.15 cells, it failed to inhibit HBV multiplication. The cytotoxicity of saikosaponin d against HepG2 human hepatocellular carcinoma cells was due to the induction of apoptosis through the activation of caspases 3 and 7, which subsequently resulted in poly-ADP-ribose-polymerase (PARP) cleavage. DNA fragmentation was clearly noted at more than 6 h after HepG2 cells exposure to saikosaponin d. The present study concludes that saikosaponin c exhibits anti-HBV activity and saikosaponin d possesses potent cytotoxicity against human hepatocellular carcinoma cells.     

Structure and actions of saikosaponins isolated from Bupleurum falcatum L. I. Anti-inflammatory action of saikosaponins.

Anti-inflammatory action of saikosaponins isolated from the root of Bupleurum falcatum L were examined using female albino rats. The anti-exudative action by granuloma pouch method and the antigranulomatous action by cotton pellet method were demonstrated with i.m. and oral administrations of saikosaponins. The oral administration of saikosaponins in 10 times the dosage of i.m. injection showed almost the same effect. Among saikosaponins isolated from Bupleurum falcatum, saikosaponins a and d, not c, were demonstrated to have anti-inflammatory action. The relationship between structure and action of saikosaponins was discussed. No changes in body weight, adrenal weight, plasma-11-OH-corticosteroid level and hematocrit value were observed.     

Reverse-phase high pressure liquid chromatographic analysis of harpagoside, scorodioside and verbascoside from Scrophularia scorodonia: quantitative determination of harpagoside

A reversed-phase high performance liquid chromatographic method has been developed for the determination of the main compounds (harpagoside, scorodioside, and verbascoside) from different samples of Scrophularia scorodonia. The chromatographic method has been validated and applied for quantitative determination of harpagoside. The results show the highest levels of harpagoside in the leaf extract. The purity and identity of peaks were controlled by diode-array detection and comparison with standards.     

New sesquiterpenoid glycoside and phenylpropanoid glycosides from the tuber of Ophiopogon japonicus

A new sesquiterpenoid glycoside, cryptomeridiol 11-O-β-d-xylopyranosyl-(1→6)-β-d- glucopyranoside (1), two new phenylpropanoid glycosides, 3,4-dihydroxy-allylbenzene 3-O-β-d-glucopyranosyl-4-O-β-d-apiofuranosyl-(1→6)-β-d-glucopyranoside (2), and 3,4,5-trihydroxy-allylbenzene 3-O-β-d-glucopyranosyl-4-O-β-d-glucopyranoside (3), along with four known phenylpropanoid glycosides (4–7), were isolated from the tuber of Ophiopogon japonicus. Compounds 4–7 were obtained from the genus Ophiopogon for the first time. Their structures were elucidated by spectroscopic methods, including 1D and 2D NMR and HR-ESI-MS.     

玄参中苯丙素苷对脱氧核苷酸羟基加成自由基的快速修复作用(英文)

目的:探讨玄参化学成分的抗氧化作用.方法:应用多种层析方法和化学和光谱方法,对玄参的水溶性主要成分进行分离和结构鉴定.应用脉冲辐解方法,观察分离得到的主要单体成分对脱氧核苷酸dAMP和dGMP羟基加成自由基的快速修复过程,测定苯丙素苷与羟基加成自由基的反应速率常数.结果:从玄参水溶性部位分得四种主要成分,发现它们属于环烯醚萜苷:哈帕酯苷与哈帕苷,和苯丙素苷:安格洛苷C与acteoside.脉冲辐解实验中观察到在0.1 mmol·L~(-1)时,苯丙素苷安格洛苷C与acteoside对脱氧核苷酸羟基加成自由基产生显著的修复作用,而环烯醚萜苷:哈帕酯苷与哈帕苷在相同条件下作用不明显.安格洛苷C与dAMP及dGMP间的电子转移速率常数为4.2×10~8及10.3×10~8 L·mol~(-1)·S~(-1);acteoside与dAMP及dGMP间的电子转移速率常数为5.3×10~8及20.2×10~8 L·mol~(-1)·S~(-1).结论:玄参中的苯丙素苷在还原脱氧核苷酸的氧化性羟基加成自由基方面有很好的抗氧化作用.     

苯丙素苷类抗氧化剂异毛蕊花苷对人胃癌细胞生长和分化的影响

目的：探讨苯丙素苷类抗氧化剂异毛蕊花苷对人胃癌MGC 803细胞生长和分化的影响。方法：以1．5％二甲基亚砜为阳性对照,采用细胞计数、肿瘤形成率实验、酶活性测试、流式细胞仪分析和Western blot等方法分别评价异毛蕊花苷对MGC 803细胞生长、致瘤性、碱性磷酸酶（ALP）及乳酸脱氢酶（LDH）活性、细胞周期和周期相关基因表达的影响。结果：MC 803细胞经异毛蕊花苷处理后,细胞生长和增殖呈浓度和时间依赖性抑制;异毛蕊花苷20μmol/L可使细胞致瘤性明显受抑,胃癌细胞分化标志酶ALP和LDH活性均呈时间依赖性下降,细胞周期表现为G0／G1期阻滞,G1→S期调控点相关蛋白p53、p21/WAF1和p16/INK4表达上调,C－myc蛋白表达受抑。结论：异毛蕊花苷可明显抑制MGC 803细胞生长及逆转胃癌细胞恶性表型进而诱导分化,这可能与细胞周期G0／G1期阻滞及其对周期相关基因表达的调控作用有关。     

Antiviral and interferon-inducing activities of a new peptidomannan, KS-2, extracted from culture mycelia of Lentinus edodes

Oral (PO) administration of KS-2 to adult DDI mice resulted in a peak serum interferon (IF) titer of 800 units (U)/ml 20 hours after administration with detectable levels persisting until 30 hours. After intraperitoneal (IP) injection, a peak serum IF titer of 1,600 U/ml was detected and it followed the same time course as that of oral administration. The IF induced by KS-2 shared certain physico-chemical properties with the standard preparation of immune IF and was not neutralized by an antiserum against type I IF. In mice infected intranasally (IN) with influenza A2 (H2N2) virus, KS-2 was found to possess significant protective activities. Efficacy of the agent was evidenced by an increase in survivor number, a prolongation of mean survival time, an inhibition of the development of lung consolidation induced by the viral infection and a decrease in virus titer in lung tissues. Both PO and IP administrations of KS-2 protected mice against infection and significant antiviral activities were achieved not only by prophylactic but also chemotherapeutic administration. The protective activities of KS-2 against influenza virus infection in mice are discussed in view of the immunopotentiation of the host animals.     

Antiviral activity of simalikalactone D, a quassinoid from Quassia africana

After removing lipophilic material, the ground root bark of Quassia africana Baill. (Simaroubaceae) was extracted with ethanol 95 %. Partitioning between chloroform, ethyl acetate and water yielded three crude extracts. Pronounced activities were shown by the chloroform and ethyl acetate crude extracts against Herpes simplex, Semliki forest, Coxsackie and Vesicular stomatitis viruses. By repeated column chromatography and preparative thin layer chromatography on silica gel, two quassinoids, i. e., quassin and simalikalactone D were isolated. Structures of the pure compounds were established primarily using NMR spectroscopy. Mass spectral information confirmed the assigned structures. Simalikalactone D was responsible, at least in part, for the high antiviral activity observed for the chloroform crude extract. Quassin showed no activity. For quassinoids the ester group at C-15 and the epoxymethano bridge between C-8 and C-13 appeared to be important structural features in order to exhibit a pronounced antiviral activity.     

In vitro anti-inflammatory activity of iridoids and triterpenoid compounds isolated from Phillyrea latifolia L

Two iridoids, oleuropeoside and ligustroside, and two triterpenoid compounds, oleanolic acid and ursolic acid, have been isolated from the leaves of Phillyrea latifolia L. (Oleaceae). These compounds were tested for interactions with the cyclooxygenase (COX) and 5-lipoxygenase (5-LOX) pathways of arachidonate metabolism in calcium ionophore-stimulated mouse peritoneal macrophages and human platelets, and for their effect on cell viability. Structure-activity relationships obtained for in vitro screening results were discussed. These compounds are capable of exerting inhibitory actions on enzymes of the arachidonate cascade. All compounds assayed showed a significant effect on prostaglandin E2 (PGE2)-release, with inhibition percentages similar to the reference drug indomethacin (IC50 = 0.95 microM). The IC50 values of the active compounds are: oleuropeoside 47 microM, ligustroside 48.53 microM, oleanolic acid 23.51 microM and ursolic acid 60.91 microM. In the leukotriene C4 (LTC4)-assay, only oleanolic acid showed a significant effect (IC50 = 16.79 microM). We also investigated the action of compounds on thromboxane B2 (TXB2)-release induced by calcium ionophore in human platelets. Of all the tested compounds, only ligustroside (IC50 = 122.63 microM) and ursolic acid (IC50 = 50.21 microM) showed a significant effect, although with less potency than the reference drug ibuprofen (IC50 = 1.27 microM). Thus, our compounds possess an array of potentially beneficial anti-inflammatory properties which may, alongside other constituents, contribute to the claimed therapeutic properties of the plant from which they are derived.     

Antiviral activity, antimetabolic activity, and cytotoxicity of 3'-substituted deoxypyrimidine nucleosides

The antiviral activity, effect on cellular DNA and RNA synthesis, and cytotoxicity toward mammalian cells of 5-fluoro-2'-deoxyuridine, 5-methoxymethyl-2'-deoxyuridine, 2'-deoxythymidine, and their corresponding 3'-p-nitrophenylphosphate and 3'-p-aminophenylphosphate derivatives were determined. The 3'-p-aminophenylphosphate-2'-deoxy-5-methoxymethyluridine derivative was as potent as 5-methoxy-methyl-2'-deoxyuridine in inhibiting herpes simplex viruses; however, 3'-p-aminophenylphosphate-2'-deoxy-5-fluorouridine was less potent than 5-fluoro-2'-deoxyuridine in inhibiting viral replication. The results suggest that the deoxypyrimidine ribonucleoside kinase has bulk tolerance for substituents at the 3-position of the ribofuranose moiety. The effect on cellular DNA and RNA synthesis and cytotoxicity toward mammalian cells were monitored by studying the incorporation of radioactive precursors. 5-Methoxymethyl-2'-deoxyuridine and 3'-p-aminophenylphosphate-2'-deoxy-5-methoxymethyluridine failed to inhibit DNA or RNA synthesis. 5-Fluoro-2'-deoxyuridine and 3'-p-aminophenylphosphate-2'-deoxy-5-fluorouridine decreased incorporation of [3H]deoxyuridine by 50% at 1.0 and 40 microM, respectively. Cytotoxicity (microscopic lesions using monolayer cells) on exposure to 5-methoxymethyl-2'-deoxyuridine, 3'-p-aminophenylphosphate-2'-deoxy-5-methoxymethyluridine, 5-fluoro-2'-deoxyuridine, and 3'-p-aminophenylphosphate-2'-deoxy-5-fluorouridine was observed at 3800, 1600, 1.6, and 110 microM, respectively.     

Anti-hepatotoxic activity of cichotyboside,a sesquiterpene glycoside from the seeds of Cichorium intybus

The seeds of Cichorium intybus L. (Asteraceae) afforded a new guaianolide sesquiterpene glycoside, cichotyboside, which was characterized as 2α, 6β, 7β, 15-tetrahydroxy-1 (10), 4 (5)-diene-guaian-9α, 12-olide-7-O-β-caffoyl-15-O-β-D-glucoside (1) by means of spectral methods. Cichotyboside (1) exhibited a significant anti-hepatotoxic activity against CCl₄ induced toxicity in Wistar rats, wherein it reduced the elevated levels of liver enzymes such as serum glutamate oxaloacetate transaminase (SGOT) by 52 units/ml; SGPT 38 units/ml; ALKP 24.97 units/ml and 7.54 g/dl, 5.48 g/dl increase in total protein and albumin, respectively. It was observed that cichotyboside (1) decreased the level of ALKP comparable with that of standard drug silymarin, exhibiting an 88% decrease in comparison to silymarin (92%) and increased the level of total albumin 85% in comparison to silymarin (89%) against intoxicated control. Whereas, the levels of SGOT and SGPT were also decreased considerably in comparison to standard and intoxicated control.     

Anti-hepatotoxic activity of cichotyboside, a sesquiterpene glycoside from the seeds of Cichorium intybus

The seeds of Cichorium intybus L. (Asteraceae) afforded a new guaianolide sesquiterpene glycoside, cichotyboside, which was characterized as 2alpha, 6beta, 7beta, 15-tetrahydroxy-1 (10), 4 (5)-diene-guaian-9alpha, 12-olide-7-O-beta-caffoyl-15-O-beta-D-glucoside (1) by means of spectral methods. Cichotyboside (1) exhibited a significant anti-hepatotoxic activity against CCl4 induced toxicity in Wistar rats, wherein it reduced the elevated levels of liver enzymes such as serum glutamate oxaloacetate transaminase (SGOT) by 52 units/ml; SGPT 38 units/ml; ALKP 24.97 units/ml and 7.54 g/dl, 5.48 g/dl increase in total protein and albumin, respectively. It was observed that cichotyboside (1) decreased the level of ALKP comparable with that of standard drug silymarin, exhibiting an 88% decrease in comparison to silymarin (92%) and increased the level of total albumin 85% in comparison to silymarin (89%) against intoxicated control. Whereas, the levels of SGOT and SGPT were also decreased considerably in comparison to standard and intoxicated control.     

管花肉苁蓉苯乙醇苷提取物对血管内皮细胞EA.hy926增殖的抑制和凋亡的诱导作用

目的 研究管花肉苁蓉苯乙醇苷提取物对血管内皮细胞EA.hy926增殖的抑制和凋亡的诱导作用.方法 采用CCK-8法测定细胞增殖的抑制率,用Hoechst33342染色法观察细胞的凋亡形态,采用Annexin V-FITC/PI双染法结合流式细胞仪测定细胞的凋亡率,用Western blot法检测凋亡相关蛋白Bcl-2、Bax的表达情况.结果 与对照组相比,管花肉苁蓉苯乙醇苷提取物能够抑制血管内皮细胞的增殖且呈剂量依赖性,其作用24h时的IC50 =30.61 μg·mL-1;能够增加血管内皮细胞凋亡率,药物组的细胞呈现核浓染和固缩现象,能够降低抗凋亡蛋白Bcl-2的表达、增加促凋亡蛋白Bax的表达、Bcl-2/Bax值显著降低.结论 肉苁蓉苯乙醇苷提取物能够通过诱导血管内皮细胞的凋亡来抑制其增殖,其机制与调控凋亡相关蛋白Bcl-2、Bax的表达有关.     

Comparative in vitro activity of LY 127935 (6059-S), seven cephalosporins, three aminoglycosides, carbenicillin, and ticarcillin

LY 127935 (6059-S), a new semi-synthetic beta-lactam antibiotic was tested simultaneously with 6 cephalosporins, 3 aminoglycosides, carbenicillin and ticarcillin against 398 clinical isolates of Gram-negative bacilli and Gram-positive cocci. Many of the organisms were selected for study because of known resistance to one or more of the clinically available antibiotics tested. Escherichia coli, Klebsiella, Serratia and Providencia were susceptible to LY 127935. Some resistant strains of Enterobacter, Proteus, Pseudomonas aeruginosa and Acinetobacter were also resistant to LY 127935, but many of the strains resistant to other antibiotics were susceptible to LY 127935. The activity of LY 127935 against Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus pyogenes, Streptococcus viridans and Streptococcus bovis was similar to that of cephalexin and cephradine. LY 127935 was not active against methicillin-resistant S. aureus nor enterococcus.     

Abortifacient and toxic actions of the glycoside ‘albitocin’ extracted from some Albizia species

Conscious intact mice, rats, guinea-pigs, rabbits and monkeys were dosed with “albitocin”, an active glycoside extracted from plants of certain Albizia species used by East African native doctors to accelerate labour and procure abortion. In pregnant animals abortion usually occurred within 12 hr at dose levels characteristic for each species irrespective of stage of gestation. In larger doses the drug was toxic, and with lethal doses animals survived 12–170 hr, with increasing apathy and anorexia, conscious but moribund as death approached. Toxicity in the orally dosed animals was lower than in those dosed intraperiotoneally or intravenously. The changes observed which could account for the mortality are discussed.     

Antiviral and antioxidant activity of flavonoids and proanthocyanidins from Crataegus sinaica

The antiviral and antioxidant activity of some fractions and of a series of flavonoids and proanthocyanidins obtained from Crataegus sinaica (Rosaceae) was evaluated. The O-glycosidic flavonoids and the oligomeric proanthocyanidins exhibited significant inhibitory activity against herpes simplex virus type 1 (HSV-1), which was shown to be due to an extracellular mechanism for procyanidin C-1. Procyanidin C-1 also had the highest antioxidant activity in both the microsomal lipid peroxidation and the hydroxyl radical scavenging assay. In addition to the previously reported phenolic compounds, the pentacyclic triterpenoid ursolic acid (1) and a tetrameric (2) and pentameric procyanidin (3) are reported for the first time.     

Antiviral activity of N-phosphorylated amides,thioamides, and their salts

Translated from Khimiko-farmatsvticheskii Zhurnal, Vol. 23, No. 5, pp. 600–602, May, 1989.