Effect of lycoriside,an acylglucosyloxy alkaloid,on mast cells

The effect of lycoriside, an acylglucosyloxy alkaloid from Crinum asiaticum Linn, (family Amaryllidaceae), with or without sitosterol-3-O--D-glucoside, was studied on the rate of degranulation of peritoneal mast cells of albino rats. Lycoriside, at lower concentrations (1–20 µg/ml), in vitro, produced statistically significant protection against Tween 80-induced degranulation, as also to sensitized mast cells challenged with an antigen (horse serum). It also provided protection against compound 48/80-induced degranulation of mast cells when administered in vivo (1–5 mg/kg, po). At higher concentrations (100 µg/ml and above), in vitro, however, it had a mast-cell degranulation effect per se. The addition of sitosterol-3-O--D-glucoside to lycoriside did not modify the effect of the latter compound. The mechanism of the dual response elicited by lycoriside is appraised in view of a concentration-dependent anti- or prerelease effect on mast-cell mediators.     

Extracts of Crinum latifolium inhibit the cell viability of mouse lymph oma cell line EL4 and induce activation of anti-tumour activity of macrophages in vitro

Ethnopharmacological relevance

Crinum latifolium L. (CL) leaf extracts have been traditionally used in Vietnam and are now used all over the world for the treatment of prostate cancer. However, the precise cellular mechanisms of the action of CL extracts remain unclear.

Aim of the study

To examine the effects of CL samples on the anti-tumour activity of peritoneal murine macrophages.

Materials and methods

The properties of three extracts (aqueous, flavonoid, alkaloid), one fraction (alkaloid), and one pure compound (6-hydroxycrinamidine) obtained from CL, were studied (i) for redox capacities (DPPH and bleaching beta-carotene assays), (ii) on murine peritoneal macrophages (MTT assay) and on lymphoma EL4-luc2 cells (luciferine assay) for cytotoxicity, (iii) on macrophage polarization (production of ROS and gene expression by PCR), and (iv) on the tumoricidal functions of murine peritoneal macrophages (lymphoma cytotoxicity by co-culture with syngeneic macrophages).

Results

The total flavonoid extract with a high antioxidant activity (IC₅₀=107.36 mg/L, DPPH assay) showed an inhibitory action on cancer cells. Alkaloid extracts inhibited the proliferation of lymphoma cells either by directly acting on tumour cells or by activating of the tumoricidal functions of syngeneic macrophages. The aqueous extract induced mRNA expression of tumour necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin 6 (IL-6) indicating differentiation of macrophages into pro-inflammatory M1 polarized macrophages. The total flavonoid, alkaloid extracts and an alkaloid fraction induced the expression of the formyl peptide receptor (FPR) on the surface of the polarized macrophages that could lead to the activation of macrophages towards the M1 phenotype. Aqueous and flavonoid extracts enhanced NADPH quinine oxido-reductase 1 (NQO1) mRNA expression in polarized macrophages which could play an important role in cancer chemoprevention. All the samples studied were non-toxic to normal living cells and the pure alkaloid tested, 6-hydroxycrinamidine, was not active in any of the models investigated.

Conclusions

Our results indicate that CL extracts and alkaloid fraction (but not pure 6-hydroxycrinamidine) inhibit the proliferation of lymphoma cells in multiple pathways. Our results are in accordance with traditional usage and encourage further studies and in vivo assays.     

Bio-guided isolation of cholinesterase inhibitors from the bulbs of Crinum x powellii

In a search for potential acetylcholinesterase (AChE) inhibitors, the ethanol extract of the bulbs of Crinum x powellii (Amaryllidaceae) was found to demonstrate a marked inhibition of this enzyme. Using a bio-guided isolation strategy, linoleic acid ethyl ester has been identified as the compound responsible for this inhibition. Three other molecules - the alkaloid hippadine, the glycosylated benzyl alcohol derivative calleryanin and 4'-hydroxy-7-methoxyflavan - were also isolated and characterized for the first time from Crinum x powellii. The structures of these compounds were elucidated by spectrometric methods including EI, D/CI mass spectrometry, (1)H, (13)C and 2D NMR experiments. Linoleic acid was also found to inhibit AChE.     

Mast cell stabilizing effect of glucan A and phosphatidyllycorine isolated from Crinum latifolium

The effect of glucan A and phosphatidyllycorine, isolated from Crinum latifolium L. (family Amaryllidaceae) was studied on the rate of degranulation of mast cells of albino rats. Different combinations of glucan A and phosphatidyllycorine (5–20 and 5–10 μg/mL, respectively) in vitro, produced statistically significant protection against Tween 80-induced degranulation, as well as to sensitized mast cells challenged with an antigen (horse serum). The combination (10–20 mg/kg), when administered in vivo, also provided protection against compound 48/80-induced degranulation of mast cells. The findings are appraised in view of the use of the total extract of C. latifolium, for the treatment of allergic disorders in Ayurvedic medicine.     

Screening of plants of Amaryllidaceae and related families from Panama as sources of acetylcholinesterase inhibitors

Context: This is the first comprehensive study of the acetylcholinesterase (AChE) inhibitory activity of species of the family Amaryllidaceae and 13 related families from Panama.

Objective: Exploration of the potential sources of AChE inhibitors with radical scavenging activity from Amaryllidaceae and 13 related families from Panama.

Materials and methods: The studied plants were screened with anti-acetylcholinesterase bioautographic and DPPH free radical scavenging assays.

Results: From the 57 plants studied, eight (14%) showed strong inhibition of AChE, and 29 (51%) plants showed moderate inhibition of AChE.

Discussion and conclusion: Sagittaria lancifolia L. (Alismataceae), Crinum jagus (Thomps.) (Amaryllidaceae), Crinum x amabile Donn (Amaryllidaceae), Crinum zeylanicum (L.) L. (Amaryllidaceae), Crocosmia x crocosmiiflora (Lemoine ex Anonymous) N.E. Br. (Iridaceae), Sisyrinchium tinctorium Kunth (Iridaceae), Agapanthus praecox subsp. orientalis (F.M. Leight.) F.M. Leight. (Liliaceae), and Xyris jupicai Rich. (Xyridaceae) were the most active plants, inhibiting AChE at 100 μg on the TLC bioautographic method for the detection of acetylcholinesterase inhibitors. Out of the eight most active plants, two plants, Crinum zeylanicum (L.) L. and Xyris jupicai Rich., showed antioxidant activity.     

Antiproliferative alkaloids from Crinum zeylanicum

Crinum zeylanicum is used in folk medicine as a rubefacient in rheumatism, a treatment for malaria or as a poison. Complex alkaloid profiles in C. zeylanicum plant organs were revealed by GC‐MS analysis, including several bioactive compounds. Crinine, lycorine, 11‐O‐acetoxyambelline, ambelline, 6‐hydroxybuphanidrine and 6‐ethoxybuphanidrine (an artefact of the isolation procedure) were isolated. Crinine, 6‐hydroxybuphanidrine and 6‐ethoxybuphanidrine showed antiproliferative effects against human tumor cell lines, crinine being the most active (IC₅₀ 14.04 μm against HL‐60/Dox). The latter compound induced apoptosis in a dose‐dependent manner in HL‐60 and MDA‐MB‐231 cell lines. Structure‐activity relationships in the studied molecules indicated that the hydrogenation of the double bond at C1‐C2 leads to a loss of activity, whereas substitutions at C6, C8 and C11 affect their cytotoxicity. Copyright © 2011 John Wiley & Sons, Ltd.     

西南文殊兰(Crinum latifalium L.)化学成分研究

目的研究西南文殊兰的化学成分。方法采用多种层析方法分离西南文殊兰乙醇提取物的化学成分,通过其化合物理化性质及各种波谱数据鉴定化合物结构。结果共分离鉴定了15个化合物,它们的结构鉴定为：-sitosterol（1）,daucosterol（2）,5-methyl-2,4（1H,3H）-pyrimidinedione（3）,-purine（4）,p-hydroxybenzonic acid（5）,ethyl 4-hydroxybenzoate（6）,cinnamyl D-glucopyranoside（7）,benzyl-D-glucopyranoside（8）,adenosine（9）,uridine（10）,Z-cinnamic acid（11）,5-O-nitrosouridine（12）,3-methyl-1H-pyrimidine-2,4-dione（13）,N-methyl-benzamide（14）,E-cinnamic acid（15）。结论化合物3~4、7~10、12~14为首次从西南文殊兰中分离得到。     

文殊兰种子中总生物碱提取方法的比较

：目的 探讨提取文殊兰Crinum asiaticum种子中总生物碱的最佳提取方法以及工艺条件。方法 以文殊兰种子为原料,采用超声和加热回流2种提取方法提取文殊兰种子中的总生物碱。结果 响应曲面法优化结果表明,超声波法超声时间47 min,超声温度58 ℃,超声功率600 W,乙醇体积分数为95%,料液比1∶6,提取3次,提取总生物碱得率0.435 7%;加热回流法提取时间2.5 h,提取温度84 ℃,料液比1∶9,乙醇体积分数为95%,提取3次,提取总生物碱得率0.449 4%。结论 综合考虑总生物碱提取率、操作方法、除杂难易程度等因素,选用加热回流提取文殊兰种子中总生物碱较理想。     

文殊兰抗肿瘤研究进展

肿瘤是威胁人类生命和生存质量的重大疾病。传统的肿瘤化疗药物表现出毒性大、选择性差和多重耐药性等缺陷,而天然药物由于资源丰富,生物活性多样,使得从天然药物中寻找效果显著、不良反应少的抗肿瘤药物成为一条重要的途径。根据国内外学者的研究证实文殊兰有着广泛的医药应用价值,现着重对文殊兰的抗肿瘤作用予以综述。     

The anti-snake venom activities of the methanolic extract of the bulb of Crinum jagus (Amaryllidaceae).

The anti-snake venom activities of the methanolic extract of the bulb of Crinum jagus plant (Amaryllidaceae) were investigated in vitro and in vivo against the venoms of three notable snake species: Echis ocellatus, Bitis arietans and Naja nigricollis. The extract was prepared by cold marceration in 50% methanol at 37 degrees C with intermittent shaking for 48 h. An yield of 12.8% w/w dry extract was obtained. Oral administration of C. jagus extract (1000 mg/kg) protected 50% of mice, while injection of a 30 min pre-incubated mixture of the same dose of extract and venom gave 100% protection against the lethal effects of E. ocellatus venom (10 mg/kg, i.m.). The intraperitoneal administration of the extract at 250 mg/kg, 30 min before the injection of E. ocellatus venom (10mg/kg, i.m.), significantly (p<0.05) prolonged the death time of poisoned mice. C. jagus extract (500 mg/kg, per os), gave 50% protection against B. arietans venom (9.5mg/kg, i.m.) in mice while the pre-incubation of a mixture of the same dose of venom and extract (500 mg/kg), prior to injection (i.p.) of the mixture, gave only 33.3% protection. The pre-incubation of 500 mg/kg of C. jagus extract with N. nigricollis venom (6 mg/kg) prior to i.p. injection of the mixture protected 50% of the treated mice. There were generally no significant differences in the death times of mice that were given the same dose of the extract orally 30 min before injection of the venoms and those administered with the pre-incubated mixtures of venom and extract. The pre-incubation of the extract and E. ocellatus venom (5mg/kg) for 30 min, before the i.m. injection of the mixture, significantly reduced infiltration of inflammatory cells to the site of injection 4h post treatment. The concentrations of plasma creatine kinase in poisoned mice were significantly (p<0.01 or p<0.05) reduced after the injection (i.p.) of C. jagus extract (1000 mg/kg) pre-incubated with E. ocellatus (5mg/kg) or B. arietans (7 mg/kg) venom, respectively. The bulb extract of C. jagus blocked the haemorrhagic activity of a standard haemorrhagic dose (2.8 mg/ml) of E. ocellatus venom at various concentrations (1.7, 3.3 and 6.7 mg/ml). The methanolic bulb extract of C. jagus was therefore able to significantly protect mice from death, myonecrosis and haemorrhage induced by the lethal effects of venoms of notable snake species in Nigeria.