Toxicological screening of Euphorbia tirucalli L.: developmental toxicity studies in rats

Euphorbia tirucalli (Euphorbiaceae family), an environmental risk factor for Burkitt's lymphoma, also presents pharmacological activities. In the northeast region of Brazil its latex is used as an antimicrobial, antiparasitic in the treatment of coughs, rheumatism, cancer and other maladies as folk remedy. The present work concerns its developmental toxicity in rats. Wistar rats on the first day post-coitum (dpc) were grouped as control (distilled water) and treated (latex aqueous solution) groups. Animals were treated by oral gavage from the 1st to the 4th (Experiment I) and from the 5th to 7th dpc (Experiment II) and killed on the 5th or 14th dpc, respectively. Maternal variables were: clinical signs of toxicity, body weight, ovaries, liver and kidneys weight and number of corpora lutea. The uterine tubes and cornua were washed for counting and identification of embryos. The embryos and placenta were weighed. Apart from the leucopenia and the higher placental weight observed in treated rats from Experiment II, there were no significant differences between the groups. It is possible to conclude that the latex aqueous solution of Euphorbia tirucalli did not interfere with tubaric embryo development or with implantation, but it seems to alter the placenta morphology.     

Tanshinlactone A from Salvia miltiorrhiza modulates interleukin-2 and interferon-gamma gene expression

Salvia miltiorrhiza Bunge (Tanshen), a traditional Chinese herbal medicine, is popularly used to treat cardiovascular disorders. In the present study, effects of tanshinlactone A (C(16)H(12)O(4); M.W. 268), newly discovered from Salvia miltiorrhiza, on phytohemagglutinin (PHA)-stimulated cell proliferation were investigated in human peripheral blood mononuclear cells (PBMC). The results indicated that tanshinlactone A inhibited PBMC proliferation activated with PHA with an IC(50) of 15.6+/-1.9 microM. Cell viability test indicated that inhibitory effects of tanshinlactone A on PBMC proliferation were not through direct cytotoxicity. Furthermore, tanshinlactone A significantly decreased the interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) gene expression in PHA-activated PBMC. It reduced the phosphorylation of mitogen-activated protein kinases (MAPK) involving extracellular signal-regulated protein kinase (ERK), P38, and c-Jun NH(2)-terminal kinase (JNK) in PHA-treated PBMC. We suggested that the inhibitory effects of tanshinlactone A on PHA-induced PBMC proliferation, appeared to be mediated, at least in part, through reduction of MAPK activation and IL-2 and IFN-gamma production. Therefore, data demonstrate for the first time that tanshinlactone A is likely an immunomodulatory agent for PBMC.     

艾纳香叶废渣乙酸乙酯部位对佐剂类风湿关节炎大鼠药理作用研究

目的:研究艾纳香叶废渣乙酸乙酯部位(BBE)对大鼠实验性佐剂类风湿关节炎的作用及其机制。方法:用弗氏完全佐剂建立佐剂型类风湿关节炎大鼠模型,用废渣乙酸乙酯部位不同剂量进行28 d的灌胃治疗处理,以体重变化、足肿胀度、关节炎指数、免疫器官增生以及病理切片等来评价治疗效果,并同时用比色法测定血清中的SOD,MDA,GSH,NO,OH·,ALP,AST,ALT,NAG,SA水平以及采用酶联免疫吸附实验(ELISA)测定血浆中IL-1,IL-6,TNF-α水平。结果:其乙酸乙酯部位能显著减轻AA大鼠关节肿胀和关节炎指数,能有效抑制关节滑膜增生,下调血清中MDA,NO,OH·,ALP,AST,ALT,NAG,SA,IL-1,IL-6,TNF-α水平,上调SOD,GSH水平。结论:艾纳香叶废渣乙酸乙酯部位对类风湿关节炎有一定的治疗作用。     

野菊花心血管活性部位化学成分的研究

目的:研究菊科植物野菊花心血管活性部位中的主要化学成分.方法:采用多种色谱方法进行分离纯化,通过波谱方法结合文献对照鉴定化合物结构.结果:从野菊花心血管活性部位中分离并鉴定了12个化合物,包括7个黄酮类化合物(1～7),5个苯丙素类化合物(8～12),分别鉴定为(2S)-圣草酚-7-O-β-D-吡喃葡萄糖醛酸苷(1),(2S)-圣草酚-7-O-β-D-吡喃葡萄糖苷(2),(2S)-橙皮素-7-O-β-D-吡喃葡萄糖醛酸苷(3),木犀草素-7-O-β-D-吡喃葡萄糖苷(4),木犀草素-7-O-β-D-吡喃葡萄糖醛酸苷(5),香叶木素-7-O-β-D-吡喃葡萄糖醛酸苷(6),槲皮素-7-O-β-D-呲喃葡萄糖苷(7),3,5-二咖啡酰奎宁酸甲酯(8),3,5-二咖啡酰奎宁酸(9),3,5-二顺式咖啡酰奎宁酸(10),1,5-二咖啡酰奎宁酸(11),1,3-二咖啡酰奎宁酸(12).结论:野菊花心血管活性部位中主要成分为黄酮类化合物,共5个化合物分别为2～3,7～8,10;共10个化合物分别为2～3,5～12系首次从该种植物分离得到.     

大果大戟挥发油化学成分

目的:研究大果大戟Euphorbia wallichii地上部分挥发油的化学成分.方法:采用水蒸气蒸馏法,得到大果大戟挥发油,利用气相色谱-质谱联用技术进行化学成分分析.结果:共分离出25个峰,用面积归一化法测定其相对质量分数,并鉴定出大果大戟地上部分挥发油中20种主要化学成分,已鉴定出的组分占挥发油总量的94.28％.结论:大果大戟地上部分挥发油化学成分包含有机酸、烷烃、萜类及其衍生物,萜类主要为倍半萜,其衍生物则主要为酮类,主要化学成分是十六烷酸(60.36％),其次是降姥鲛-2-酮(7.674％)和十四烷酸(5.433％).     

中药十八反“藻戟遂芫俱战草”的毒理学研究进展

中药“十八反”是在相反基础上形成的一组严格的配伍禁忌,然而从古至今临床应用反药治疗疾病的例子却层出不穷.反药是否相反,现代药性理论争议很多,并且也尚未形成较统一的判断标准和结论.通过查阅文献,从临床应用、制剂及给药方式、配伍比例、对P450酶系影响和毒理方面介绍了“十八反”中有关“海藻、大戟、甘遂、芫花反甘草”的研究现状,指出以往“藻戟遂芫俱战草”的研究,主要集中在急性毒性、长期毒性,对特定部位的实验研究还很少,并且存在药物基源、给药方式、配伍比例等实验条件不统一的现象,在此基础上提出应规范实验条件,降低各种不确定因素的影响,还应根据不同药物的作用特点从特定部位对海藻、大戟、甘遂、芫花配伍甘草进行系统研究,以期为藻戟遂芫伍甘草的准确用药和进一步深入研究提供文献基础.     

醋制降低京大戟对人正常肝细胞LO2的毒性及机制研究

目的:比较京大戟醋制前后对人正常肝细胞LO2的毒性作用,并探讨其可能的作用机制.方法:体外培养LO2细胞,用不同浓度的京大戟生品和醋品对其进行处理,MTT法测定各样品对LO2细胞增殖的抑制作用;Hoeehst 33258荧光染色法观察细胞凋亡的形态学改变;AnnexinV/PI染色流式细胞术检测LO2细胞凋亡;PI染色流式细胞术分析其对细胞周期的影响;检测细胞培养液上清谷丙转氨酶(ALT)、谷草转氨酶(AST)、乳酸脱氢酶(LDH)活力,细胞裂解上清超氧化歧化酶(SOD)活力、丙二醛(MDA)和谷胱甘肽(GSH)含量.结果:与阴性对照组比较,京大戟生品各浓度组均可抑制LO2细胞增殖,诱导细胞凋亡,引起细胞S期阻滞(P＜0.05);能显著升高细胞培养液上清ALT,AST,LDH活力(P＜0.05),显著降低细胞裂解上清SOD活性和GSH含量、增加MDA含量.醋制后,与京大戟生品组比较,可显著降低细胞凋亡率、降低细胞周期阻滞,显著降低细胞培养液上清ALT,AST,LDH活力(P＜0.05),显著升高细胞裂解上清SOD活性和GSH含量、显著降低MDA含量.结论:醋制可降低京大戟对LO2细胞的毒性,其机制可能是减轻氧化损伤,从而降低细胞周期阻滞,减少细胞凋亡.     

In vivo anti-tussive activity and structural features of a polysaccharide fraction from water extracted Withania somnifera

Aim of the study

Anti-tussive drugs are amongst the most widely used medications worldwide; however no new class of drugs has been introduced into the market for many years. The present study aims at evaluating the structural features and in vivo anti-tussive activity of a polysaccharide fraction from water extracted Withania somnifera.

Materials and methods

Herein, we have analyzed water extracted material of Withania somnifera using chemical, chromatographic, spectroscopic and biological methods.

Results

A polysaccharide fraction (F3) containing arabinosyl, galactosyl and galacturonosyl residues were obtained by anion exchange chromatography of the water extracted material. This polymer is branched and contained (1,5)-/(1,3,5)-linked arabinofuranosyl, (1,3)-/(1,6)-/(1,3,6)-linked galactopyranosyl residues together with small amount of terminal rhamnopyranosyl and terminal arabinofuranosyl residues. Peroral administration of this pectic arabinogalactan in a dose of 50 mg kg⁻¹ body weight (b.w.) decreased the number of cough efforts induced by citric acid in guinea pigs like that of codeine.

Conclusions

This study provides a scientific basis for the past and present ethnomedical uses of this plant.     

Evaluation of antiulcer activity of chromanone fraction from Calophyllum brasiliesnse Camb

Ethnopharmacological relevance

Calophyllum brasiliense Camb. (Clusiaceae), popularly known as ‘guanandi’, is found in the tropical areas and swampy lands. The latex exuding from its bark is used in the treatment of gastric ulcer in folk medicine. Several active compounds have been isolated from its stem bark among them, are the chromanone acids. Therefore, this study aimed to evaluate antiulcer activity and probable mechanism(s) of action of a fraction containing a mixture of chromanone acids (BI), derived by column chromatography fractionation of the hexane extract of the stem bark of Calophyllum brasiliense (HECb), using experimental in vitro and in vivo models.

Materials and methods

Ulcer was induced by oral administration of ethanol (75%, v/v) and indomethacin (50 mg/kg). Malondialdehyde, reduced glutathione and catalase activity was measured in stomach tissue after ethanol induced ulcer. In order to evaluate the effect of BI on nitric oxide, ulcer was induced by ethanol in l-NAME pretreated animals. Anti-Helicobacter pylori activity was verified in disk diffusion and broth microdilution in vitro assays, using cagA+ and vacA+ Helicobacter pylori strains.

Results

BI prevented the gastric ulceration caused by ethanol and indomethacin treatments. Its gastroprotective mechanism in ethanol-induced ulcer was partly due to reduction of MDA and CAT levels in the gastric tissue. BI did not affect the GSH levels and its gastroprotective effect was not reversed by pretreatment with l-NAME. BI showed anti-Helicobacter pylori in the both assays.

Conclusion

The results indicate that BI is partly responsible for the HECb antiulcer and anti-Helicobacter pylori effects.     

Anti-nociceptive activity of aqueous fraction from the MeOH extracts of Paederia scandens in mice

AIM OF THE STUDY: We examined the effects of the aqueous fraction (AF) on nociception models mice induced by the chemical and the thermal stimuli so as to elucidate the analgesic activity and provide scientific basis for the clinical use of Paederia scandens. MATERIALS AND METHODS: The AF of MeOH extract from P. scandens was evaluated on anti-nociceptive activity in mice using chemical and thermal models of nociception. RESULTS: Given orally, the aqueous fraction at doses of 200, 400 and 800 mg/kg produced significant inhibitions on chemical nociception induced by intraperitoneal acetic acid and subplantar formalin injections and on thermal nociception in the tail-flick test and in the hot plate test. More significant inhibition of nociception was observed at dose of 800 mg/kg of this fraction. In the pentobarbital sodium -induced sleeping time test and the open-field test, the aqueous fraction neither significantly enhanced the pentobarbital sodium -induced sleeping time nor impaired the motor performance, indicating that the observed anti-nociception was unlikely due to sedation or motor abnormality. CONCLUSIONS: These results suggested that the aqueous fraction produced anti-nociception possibly related to the iridoid glycosides and polysaccharides in this fraction.     

中药地锦草酚性成分

目的:为研究中药地锦草抗HBV活性物质基础。方法:采用聚酰胺、大孔树脂、Sephadex LH-20,MCI GEL CHP20P等色谱方法,从地锦草70%乙醇提取物中分离得到7个酚性化合物。结果:利用MS,NMR等波谱分析手段鉴定了化合物的结构,分别为:短叶苏木酚(brevifolin,1),短叶苏木酚酸(brevifolin carboxylic acid,2),短叶苏木酚酸甲酯(methyl brevifolin-carboxylate,3),phyllanthussin E methyl ester(4),地榆酸双内酯(sanguisorbic acid dilactone,5),3,3′-二甲氧基鞣花酸(3,3′-2-di-O-methyl ellagic acid,6),鞣花酸(ellagic acid,7)。结论:化合物2~6为首次从该植物中分得。     

准噶尔大戟的化学成分研究

目的 对准噶尔大戟中的化学成分进行分离鉴定.方法 采用硅胶、Sephadex LH-20及MPLC制备色谱技术,通过理化方法和光谱分析鉴定其结构.结果 从准噶尔大戟的乙醇提取物中得到11个化合物,分别鉴定为香茅二醇(Ⅰ)、白桦脂醇(Ⅱ)、白桦脂酸(Ⅲ)、齐墩果酸(Ⅳ)、7-O-β谷甾醇(Ⅴ)、3-甲氧基-4-羟基反式苯丙烯酸正二十八醇酯(Ⅵ)、二十八酸(Ⅶ)、I-二十八烯(Ⅷ)、24-亚甲基-环阿尔廷醇(Ⅸ)、大戟醇(Ⅹ)、β-谷甾醇(Ⅺ).结论 化合物Ⅰ～Ⅷ均为首次从该植物中分离得到.     

Anti-inflammatory activity of flavonoids from Eupatorium arnottianum

Three anti-inflammatory compounds: nepetin, jaceosidin and hispidulin have been isolated and identified from Eupatorium arnottianum Griseb. dichloromethane extract. Nepetin reduced the TPA mouse ear edema by 46.9% and jaceosidin by 23.2% (1mg/ear). Both compounds inhibited the NF kappaB induction by 91 and 77%, respectively. Furthermore phytochemical analysis of the ethanol extract has led to the identification of eriodictyol, hyperoside, rutin, caffeic and chlorogenic acids. All these compounds are reported for the first time in this species. The finding of topical antiinflammatory activity exerted by Eupatorium arnottianum extract and the identification of active principles could support the use of this plant for the treatment of inflammatory affections.     

The production of nitric oxide and prostaglandin E2 in peritoneal macrophages is inhibited by Andrographis paniculata, Angelica sinensis and Morus alba ethyl acetate fractions

Aim of the study

Traditional Chinese medicine herbs (TCMHs) are used in medicines as well as in daily dietary supplements in Asia. In this study, we employed pNF-κB-Luc or pIFN-γ-Luc and BALB/c mice peritoneal macrophages or splenocytes to investigate both the immune and inflammatory effects of six selected plant species.

Materials and Methods

Specifically, we used ethyl acetate fractions of Astragalus membranaceus (Fisch.) Bunge var. mongholicus (Bunge) Hsiao (Fabaceae) (AM), Andrographis paniculata (Burm. f.) Nees (Acanthaceae) (AP), Angelica sinensis (Oliv.) Diels (Apiaceae) (AS), Eucommia ulmodes Oliv. (Eucommiaceae) leaves (EU leaves), Isatis indigotica Fort. (Brassicaceae) (II) and Morus alba L. (Moraceae) (MA).

Results

We found that ethyl acetate fractions of AP, AS and MA significantly decreased NF-κB luciferase activity and also the secretion of NO and PGE₂ in LPS/IFN-γ stimulated mouse peritoneal macrophages (p < 0.05). In contrast, they did not affect IFN-γ luciferase activity or IFN-γ production in concanavalin A (Con A)-activated mouse splenocytes. Our results indicated that the anti-inflammatory properties of these plant extracts might be resulted from the inhibition of pro-inflammatory mediators (e.g., NO and PGE₂), at least in part via suppression of a signaling pathway such as NF-κB.

Conclusions

Collectively, we have found that three potent bioactive TCMH species exerted significant NF-κB inhibitory activity and acted in a cell type dependent fashion.     

Anti-diabetic potential of alkaloid rich fraction from Capparis decidua on diabetic mice

Ethno pharmacological relevance

Capparis decidua (CD) is a xerophytic shrub, found widely in the western parts of India, Pakistan and some of the Asian countries. The dried fruits are used as an ingredient in anti-diabetic compositions.

Aim of the study

The present study was carried out to test the effect of alkaloid rich (AR) fraction from this plant in the management of diabetes.

Materials and methods

Streptozotocin-induced diabetic mice were treated with the AR fraction for 28 days. On completion of the treatment, a range of parameters were tested including oral glucose tolerance test (OGTT), blood lipid profile, expression patterns of various glucose homeostatic enzyme genes and their activities.

Results

Treatment of diabetic mice with AR fraction for 28 days significantly inhibited the acute elevation of blood glucose level during OGTT and also reduced total cholesterol (TC) and triglyceride (TG) content (p < 0.05). Activity of glucose-6-phosphatase (G6Pase) was attenuated by 44%, also liver and muscle glycogen content showed significant improvement (p < 0.05). The expression of different target genes like G6Pase, phosphoenolpyruvate carboxykinase (PEPCK), aldose reductase and tumor necrosis factor-α (TNF-α) showed significant reduction whereas glucose transporter-4 (Glut-4), peroxisome proliferator activated receptor-γ (PPAR-γ) and glucokinase (GK) improved remarkably.

Conclusions

AR fraction showed promising results in terms of anti-diabetic activities establishing its candidacy for further purification and characterization of the individual alkaloids, in order to understand their mechanism of action.     

Antiplasmodial activity of extracts from seven medicinal plants used in malaria treatment in Cameroon

Aim of the study

In a search for new plant-derived biologically active compounds against malaria parasites, we have carried out an ethnopharmacological study to evaluate the susceptibility of cultured Plasmodium falciparum to extracts and fractions from seven Cameroonian medicinal plants used in malaria treatment. We have also explored the inhibition of the Plasmodium falciparum cysteine protease Falcipain-2.

Materials and methods

Plant materials were extracted by maceration in organic solvents, and subsequently partitioned or fractionated to afford test fractions. The susceptibility of erythrocytes and the W2 strain of Plasmodium falciparum to plant extracts was evaluated in culture. In addition, the ability of annonaceous extracts to inhibit recombinant cysteine protease Falcipain-2 was also assessed.

Results and discussion

The extracts showed no toxicity against erythrocytes. The majority of plant extracts were highly active against Plasmodium falciparumin vitro, with IC₅₀ values lower than 5 μg/ml. Annonaceous extracts (acetogenin-rich fractions and interface precipitates) exhibited the highest potency. Some of these extracts exhibited modest inhibition of Falcipain-2.

Conclusion

These results support continued investigation of components of traditional medicines as potential new antimalarial agents.     

醋狼毒饮片质量标准研究

目的:制订醋狼毒饮片的质量标准。方法:收集炮制10批醋狼毒饮片,对水分、总灰分、酸不溶性灰分、浸出物进行测定,并以狼毒乙素含量为指标进行含量测定。结果:拟定醋狼毒饮片水分不得超过9.12%,总灰分不得超过6.50%,酸不溶性灰分不得超过1.09%,醇溶性浸出物不少于7.70%,狼毒乙素含量不少于0.038%。结论:初步建立醋狼毒饮片量化质控标准评价体系。     

Protective effects of butanol fraction from Betula platyphyla var. japonica on cartilage alterations in a rabbit collagenase-induced osteoarthritis

Aim of this study

Many cartilage protective agents have been developed from natural products, and they have resulted in the development of treatments for osteoarthritis. In this study, we determined the osteoarthritic efficacy and mechanism of butanol fraction from the bark of Betula platyphylla var. japonica (BFBP) in collagenase-induced rabbit model of osteoarthritis (CIA).

Materials and methods

The right knees of rabbits were injected intra-articularly with collagenase, and rabbits were orally administrated with distilled water (vehicle), BFBP (50, 100 and 200 mg/kg) or celecoxib (100 mg/kg) once a day for 28 days after the initiation of the CIA.

Results

Oral administration of BFBP dose-dependently suppressed the stiffness and global histologic score. Proteoglycan intensity was considerably increased in a dose-dependent manner. As well, the mRNA expression of MMP-1, and MMP-3 was decreased. On the contrary, the level of TIMP-1 in the synovial fluids was significantly increased in the BFBP treated group. The pathologic inflammatory molecules such as PGE₂ and COX-2 were inhibited by BFBP, but COX-1 expression not affected.

Conclusion

We suggest that BFBP has shown the protective effect on cartilage alternations through balance of MMPs/TIMP-1 and regulates inflammatory-related molecules in vivo model of osteoarthritis, and great candidate for development of osteoarthritis treatment.     

Sesquiterpene lactones from Gynoxys verrucosa and their anti-MRSA activity

Ethnopharmacological relevance

Because of its virulence and antibiotic resistance, Staphylococcus aureus is a more formidable pathogen now than at any time since the pre-antibiotic era. In an effort to identify and develop novel antimicrobial agents with activity against this pathogen, we have examined Gynoxys verrucosa Wedd (Asteraceae), an herb used in traditional medicine in southern Ecuador for the treatment and healing of wounds.

Materials and methods

The sesquiterpene lactones leucodine (1) and dehydroleucodine (2) were extracted and purified from the aerial parts of Gynoxys verrucosa, and their structure was elucidated by spectroscopic methods and single-crystal X-ray analysis. The in vitro anti-microbial activity of Gynoxys verrucosa extracts and its purified constituents was determined against six clinical isolates including Staphylococcus aureus and Staphylococcus epidermidis strains with different drug-resistance profiles, using the microtiter broth method.

Results

Compound 1 has very low activity, while compound 2 has moderate activity with MIC₅₀s between 49 and 195 μg/mL. The extract of Gynoxys verrucosa has weak activity with MIC₅₀s between 908 and 3290 μg/mL.

Conclusions

We are reporting the full assignment of the ¹H NMR and ¹³C NMR of both compounds, and the crystal structure of compound 2, for the first time. Moreover, the fact that compound 2 has antimicrobial activity and compound 1 does not, demonstrates that the exocyclic conjugated methylene in the lactone ring is essential for the antimicrobial activity of these sesquiterpene lactones. However, the weak activity observed for the plant extracts, does not explain the use of Gynoxys verrucosa in traditional medicine for the treatment of wounds and skin infections.     

Immunosuppressive activity of ethanolic extract of seeds of Moringa oleifera Lam. in experimental immune inflammation

Ethnopharmacological relevance

Traditionally, the plant Moringa oleifera Lam. (Moringaceae) is used for the treatment of ascites and rheumatism, while the dried seeds of the plant are used as an ‘anti-allergic’ agent.

Aim of the study

The aim of the study was to assess the efficacy of ethanolic extract of seeds from Moringa oleifera Lam. in experimental immune inflammation.

Materials and methods

Circulatory and splenic leukocyte counts, delayed-type hypersensitivity reactions and humoral antibody responses were measured in mice using SRBC as the antigen. In addition, macrophage phagocytosis was measured by the carbon clearance test.

Results

The extract dose-dependently (50, 100 and 200 mg/kg) inhibited spleen weight as well as circulatory leukocyte and splenocyte counts. The delayed-type hypersensitivity reaction was significantly inhibited (P < 0.01) by decreasing the mean foot pad thickness at 48 h. The production of the humoral antibody titer was significantly ameliorated at a dose of 100 and 200 mg/kg (P < 0.05 and P < 0.01, respectively). Furthermore, the extract caused a down-regulation of macrophage phagocytosis due to carbon particles.

Conclusion

Taken together, the above findings suggest that the seeds of Moringa oleifera have immunosuppressive activity.