Correlation study on chemical constitutes of cardiac glycosides in Taxillus chinensis and its Nerium indicum host by UPLC-Q-TOF-MS/MS / 中国中药杂志

To build up an identification method on cardiac glycosides in Taxillus chinensis and its Nerium indicum host, and evaluate the influence on medicine quality from host to T. chinensis, ultra-performance liquid chromatography coupled with quadrupole-time-of-flight tandem mass-mass spectrometry(UPLC-Q-TOF-MS/MS)was applied. The samples of T. chinensis(harvested from N. indicum)and its N. indicum host were collected in field. The samples of T. chinensis(harvested from Morus alba)and its M. alba host was taken as control substance. All samples were extracted by ultrasonic extraction in 70% ethanol. Chromatographic separation was performed on an ACQUITY UPLC HSS T3 C_(18)(2.1 mm×100 mm,1.8 μm)column at 40 ℃. Gradient elution was applied, and the mobile phase was consisted of 0.1% formic acid water and acetonitrile. The 0.5 μL of sample solution was injected and the flow rate of the mobile phase was kept at 0.6 mL·min~(-1) in each run. It was done to identify cardiac glycosides and explore the chemical composition correlation in T. chinensis and its N. indicum host by analyzing positive and negative ion mode mass spectrometry data, elemental composition, cardiac glycoside reference substance and searching related literatures. A total of 29 cardiac glycosides were identified, 28 of it belonged to N. indicum host, 5 belonged to T. chinensis(harvested from N. indicum host), none of cardiac glycoside was identified in T. chinensis(harvested from M. alba host). The result could provide a reference in evaluating the influence in T. chinensis medicine quality from host. It was rapid, accurate, and comprehensive to identify cardiac glycosides in T. chinensis and its N. indicum host by UPLC-Q-TOF-MS/MS.

Influence of traditional medicines on the activity of keratinocytes in wound healing: an in-vitro study / 대한해부학회지

Natural medicinal systems such as Ayurveda and folk medicine has remedies for wound management. However, the exact cellular and extracellular mechanisms involved in the healing process and its influence on keratinocytes is less discussed. Therefore, the present study was designed to evaluate the effect of certain natural wound healing medicines on the biology of the keratinocytes/HaCaT cells. Test materials such as honey (H), ghee (G), aqueous extracts of roots of Glycyrrhiza glabra (GG) and leaves of Nerium indicum (NI) were considered. The HaCaT cells were treated with the test materials singly and in combinations (H+G, all combined [Tot]) for a specific period (24, 48, and 72 hours). The cells were then subjected to cytotoxicity/proliferation and migration/scratch assays. All the test materials, except NI, were non-cytotoxic and showed increased cell proliferation at variable concentrations. Significant observations were made in the groups treated with honey (100 µg/ml at 48 hours, P<0.05; 1,000 µg/ml at 72 hours, P<0.05), GG (all concentrations at 48 hours, P<0.05; 750 µg/ml at 72 hours, P<0.05), H+G (250 µg/ml at 24 hours, P<0.001; 500 µg/ml at 48 and 72 hours, P<0.05), and Tot (50 µg/ml at 24, 48 and 72 hours, P<0.01). In the in-vitro wound healing assay, all the treated groups showed significant migration and narrowing of the scratch area by 24 and 48 hours (P<0.001) compared to control. The results obtained from the present study signifies the positive influence of these natural wound healing compounds on keratinocytes/HaCaT cells.

Improved method for diagnosis of Nerium oleander poisoning in necropsy tissues / Método aprimorado para diagnóstico da intoxicação por Nerium oleander em tecidos de necropsia

Nerium oleander is an ornamental cardiotoxic plant found in tropical and subtropical areas of the World. Its toxicity is related to the content of cardioactive glycosides, mainly oleandrin, found throughout the plant. The present study aimed to describe a new and improved method for oleandrin detection in tissue samples. The determination of oleandrin was made after extraction with a modified QuEChERS technique and measurement by UFLC-MS/MS. A total of 36 guinea pigs (Cavia porcellus) were distributed into 3 groups (n=12): control group that received only water orally (CON), and two treated groups that received hydroalcoholic oleander extract at doses of 150mg.kg-1 (OLE 150) and 300mg.kg-1 (OLE 300) in single oral dose. After three hours, fragments of heart, kidneys, liver and brain were collected for determination of oleandrin levels. The extraction and chromatographic procedures were effective for oleandrin detection and quantification in tissues, with retention time of 1.2 min and detection limit of 0.001μg g-1. The chromatographic analysis of treated guinea pigs indicated that oleandrin is distributed equally among the analyzed tissues. The developed methodology is a reliable, effective and rapid form of diagnosis of N. oleander poisoning based on necropsy tissue samples.(AU)

Nerium oleander é uma planta cardiotóxica ornamental encontrada em áreas tropicais e subtropicais do mundo. Sua toxicidade é relacionada á presença de glicosídeos cardioativos, principalmente a oleandrina, encontrada em toda a planta. O presente estudo objetiva descrever um novo e aprimorado método para detecção da oleandrina em amostras de tecido. A determinação da oleandrina foi feita após extração utilizando técnica modificada de QuEChERS e mensuração por UFLC-MS/MS. Um total de 36 cobaios (Cavia porcellus) foi distribuído em três grupos (n=12): grupo controle que recebeu apenas água por via oral (CON), e dois grupos tratados que receberam extrato hidroalcóolico de oleander nas doses de 150mg.kg-1 (OLE 150) e 300mg.kg-1 (OLE 300) em uma única dose oral. Após três horas, fragmentos do coração, rins, fígado e cérebro foram coletados para determinação dos níveis de oleandrina. A extração e procedimentos cromatográficos foram eficientes na detecção e quantificação da oleandrina nos tecidos, com tempo de retenção de 1,2min e limite de detecção de 0,001μg g-1. A análise cromatográfica dos animais tratados indicou que a oleandrina é distribuída de forma equalizada pelos tecidos analisados. A metodologia desenvolvida representa uma forma de diagnóstica segura, efetiva e rápida da intoxicação por N. oleander a partir de amostras de tecidos de necropsia.(AU)

Cytotoxic Triterpenoids from the Fruits of Ligustrum japonicum

Medicinal plants are potential sources of anticancer agents screening. A large number of phytochemicals, including triterpenoids, have been reported to have significant cytotoxic effects on cancer cells. From the fruits of Ligustrum japonicum Thunb., thirteen triterpenoids (1 – 13) were isolated and evaluated for their cytotoxic activity against Hela and HL-60 cells. As results, 8 (oleanolic acid) showed significant effects on Hela with IC50 values of 5.5 µM, and moderate effects on HL-60 cells with IC₅₀ values of 55.9 µM. Meanwhile, 10 (oleanderic acid) and 11 (3β-acetoxy-urs-12-en-28-oic acid) exhibited moderate inhibitory effects on Hela with IC₅₀ value of 55.0 and 68.8 µM, respectively. Moreover, 10 showed cytotoxic effect on HL-60 cell line with IC₅₀ value of 63.9 µM. To our knowledge, this is the first report that oleanderic acid was isolated from L. japonicum and investigated in cytotoxic effects on Hela and HL-60 cells.

[Effects of Nerium oleander leaf, Ricinus communis oil, Capsicum spp. seeds, and almond compound on cutaneous leishmaniasis caused by Leishmania species under laboratory condition and its effect on cutaneous lesion progression in mice]

Protozoan parasite of genus Leishmania is the causative agent of leishmaniasis in humans, carnivorous animals and rodents. In Iran, Leishmania species are causes of cutaneous leishmaniasis which had been reported frequently from Isfahan. The aim of the present study was to evaluate the effects of a compound of Nerium oleander leaf, Ricinus communis oil, Capsicum spp. seeds, and almond on cutaneous leishmaniasis caused by Leishmania species under laboratory conditions and its effect on the progression of cutaneous lesion in mice. In this repeated experimental study, we obtained smears from the skin lesions of 10 patients with cutaneous leishmaniasis randomly. 3 cases had positive smears [30%] which were cultured in NNN media with 10% FBS to obtain promastigotes of leishmania. The effects of boiled and sieved herbal compound [Nerium oleander leaf, Ricinus communis oil, Capsicum spp. seeds, and almond] on the promastigote propagation in NNN media with 10% FBS [5x10[4]/micro l] were investigated in triplicate and compared with control group. In the 4th week, 0.2 ml of enriched promastigote of Leishmania in RPMI was inoculated in the base of the tails of the mice in the test group [n=6] and control group [n=2]. The effect of herbal compound on the progression of the lesion was investigated for 7 weeks. We used t-test for statistical analysis. The number of Leishmania promastigotes in the group treated with herbal compound was significantly less than that in the control group. Also the herbal compound had significant preventive effect on the progression of the cutaneous lesion in the test group as compared to the control group. The results of this study showed that the herbal compound had preventive effect on the parasite propagation and progression of cutaneous lesion. We recommend furthur studies on other pharmacological aspects of this herbal compound

Hepatoprotective and antioxidant activity of methanolic extract of flowers of Nerium oleander against CCl4-induced liver injury in rats

OBJECTIVE@#To investigate the antioxidant and hepatoprotective activity of methanolic flower extract of Nerium oleander against CCl(4)-induced hepatotoxicity in rats.@*METHODS@#In vitro antioxidant activity of methanolic extract of flowers of Nerium oleander (MENO-F) was evaluated by various assays, including reducing power, lipid peroxidation, DPPH, ABTS, superoxide anion, hydroxyl radicals and metal chelation. The hepatoprotective and in vivo antioxidant activity of MENO-F were evaluated against CCl(4)-induced hepatic damage in rats. The MENO-F at dose of 100, 200 and 400 mg/kg were administered orally once daily for seven days. Serum enzymatic levels of serum glutamate oxaloacetate transaminase (AST), serum glutamate pyruvate transaminase (ALT), serum alkaline phosphatase (ALP) and total bilirubin were estimated along with estimation of superoxide dismutase (SOD) and malondialdehyde (MDA) levels in liver tissues. Further histopathological examination of the liver sections was carried out to support the induction of hepatotoxicity and hepatoprotective efficacy.@*RESULTS@#The extract showed potent activities on reducing power, lipid peroxide, DPPH, ABTS, superoxide anion, hydroxyl radical and metal chelation. The substantially elevated serum enzymatic levels of AST, ALT, ALP and total bilirubin were found to be restored towards normalization significantly by the MENO-F in a dose dependent manner with maximum hepatoprotection at 400 mg/kg dose level. The histopathological observations supported the biochemical evidences of hepatoprotection. Elevated level of SOD and decreased level of MDA further strengthen the hepatoprotective observations.@*CONCLUSIONS@#The results of the present study strongly reveal that MENO-F has potent antioxidant activity and hepatoprotective activity against CCl(4)-induced hepatic damage in experimental animals.

Digitalis-like Toxic Symptoms Occurring after Accidental Nerium indicum Poisoning

Although Nerium indicum poisoning is a globally rare occurrence, Nerium oleander poisoning is known to occur frequently in the Mediterranean regions. To our knowledge, this is the first reported case of accidental Nerium indicum poisoning in Korea. Its poisoning symptoms and signs are similar to that of digitalis poisoning, because of the presence of cardiac glycosides in Nerium indicum. A 16-year-old boy was admitted to the emergency department four hours prior to the accidental ingestion of Nerium indicum petals. The patient complained of nausea, vomiting, and dizziness. His initial vital signs were stable; laboratory blood test results were within normal levels, except for the blood digoxin level (1.5 ng/dL). An electrocardiogram (ECG) analysis showed normal sinus rhythm, progressive PR prolongation and second-degree Morbiz type I AV block. Conservative treatments including activated charcoal administration were conducted, because toxic symptoms and signs were not severe. The patient was admitted to the intensive care unit for close observation. His ECG was converted to normal rhythm after 1 day and the toxic symptoms and signs were completely resolved after 4 days.

Air pollution effects on the activity of antioxidant enzymes in Nerium oleander and Robinia pseudo acacia plants in Tehran

The air pollution effects on the activity of antioxidant enzymes were investigated on Nerium oleander and Robinia pseudo acacia in Tehran. Considering the information obtained from the Department of the Environment of Iran, Sorkh Hesar Park as well as South Azadi were selected as two sampling sites representing the unpolluted and polluted area respectively. A number of plant leave samples were collected from both sampling sites simultaneously. The activity of plant enzymes including peroxidase, catalase and ascorbate peroxidase was investigated using spectrophotometric methods. A higher level of peroxidase and catalase enzymes were measured in both plant samples collected from polluted area. However, this higher level was only statistically significant for the activity of peroxidase enzyme in Robinia pseudo acacia plants compare of to the control group [p<0.05]. The lower level of ascorbate peroxidase was observed in Nerium oleander plant leaves collected from the contaminated sampling site [p<0.05], but though, the activity of this enzyme in Robinia pseudo acacia did not change significantly. The overall plant injury symptoms found in this study demonstrated that both Nerium oleander and Robinia pseudo acacia have a potential to be considered as effective bioindicators to reflect the environmental air quality in polluted areas

Molluscicidal and physiological activities of some plants against biomphalaria Alexandria snails

The molluscicidal activity of three plants: Nerium oleander, Dizygotheca elegantissima and A gave decipiens on Biomphalaria Alexandrina snails at different exposure intervals and size or age of snails was studied throughout the present investigation. Also, the effects of these selected plants on Schistosoma mansoni cercariae and miracidia were assessed. Moreover, the role of sublethal concentrations of these plants on physiological processes and functions on the hemolymph of these snails was also clarified. Results showed that N. oleander proved the most effective plant as its LC[90] was 100ppm after 24hrs from exposure compared with the values of 120 and 180 ppm with the plants D. elegantissima and A. decipiens, respectively. The elongation of remarkable exposure periods from 24 to 48 hrs increased the molluscicidal activities of the tested plants. Different sizes or ages of Biomphalaria Alexandrina snails showed different susceptibility levels towards the action of the tested plants. Newly hatched snails were the most sensitive group. When the LC[50] and LC[90] of the experimental plants were tested against the cercariae, almost 100% of cercarial mortality was obtained after only two hrs when used 100, 120 and180 ppm for Nerium oleander, Dizygotheca elegantissima and Agave decipiens, respectively. Meanwhile, 100% miracidia mortality was obtained after 1 hr when exposed to the same concentrations. Thus miracidia proved more susceptible than cercariae. While used the sublethal concentrations of these plants, the LC[0] of the three tested plants showed moderate significant increase in transaminases activities. The total protein, albumin and globulin were slightly changed; while the A/G ratio did not record any changes between treated and control snails. The application of plant molluseicides resulted in marked increase of total lipid content; the maximal observation was recorded with the use of both N. oleander, and D. elegantissima

Molluscicidal activity of Nerium indicum Mill, Pterocarya stenoptera DC, and Rumex japonicum houtt on Oncomelania hupensis / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To evaluate the molluscicidal activities of three Chinese plants N. indicum Mill, R stenoptera DC, and R. japonicum Houtt, and to clarify the molluscicidal mechanism.</p><p><b>METHODS</b>N-butanol extracts and water extracts of the three plants were obtained. The reactions of EST isozyme, glycogen and total protein of snails to the plant extracts were studied.</p><p><b>RESULTS</b>EST electrophoresis showed that EST was an important antidotal enzyme system and reacted strongly to environment. EST changed greatly during the whole exposure period so that it could be viewed as a pathological index of toxicity. Extracts decreased the glycogen content of the snails' soft tissues greatly, and also the protein content.</p><p><b>CONCLUSION</b>All extracts show strong molluscicidal activity. The LD50 value of the water extract of N. indicum Mill is as low as 13.2 mg/L. EST can be viewed as a pathological index of toxicity. The energy metabolism abnormity is the key reason for the molluscicidal activities. The biochemical mechanism needs further research.</p>

Larvicidal potential of Nerium indicum and Thuja oriertelis extracts against malaria and Japanese encephalitis vector.

Ethanolic and acetone extracts of Nerium indicum and Thuja orientelis have been studied against III instar larvae of Anopheles stephensi and Culex quinquefasciatus. Ethanolic extract of N. indicum is found more effective than its acetone extract against anopheline larvae with LC50 values of 185.99 and 148.05 ppm for former and 229.28 and 149.43 ppm for the later after 24 and 48 hrs of exposure. The acetone extract with LC50 values of 209.00 and 155.97 ppm is more effective in case of culicine larvae than its ethanolic extract with LC50 494.07 and 194.49 ppm after 24 and 48 hours of treatment. Ethanolic extract of T. orientelis is more effective against both the larval species with LC50 values of 13.10 and 9.02 ppm after 24 and 48 hours for anopheline and 22.74 and 16.72 ppm against culicine larvae. The acetone extract showed LC50 values of 200.87 and 127.53 ppm against anopheline and 69.03 and 51.14 ppm against culicine larvae. Thus ethanolic extract of T. orientelis is an ideal potential larvicide for both types of mosquito larvae.

Toxicity of Nerium indicum Miller seed extract on bandicoot rat, Bandicota bengalensis Gray.

Crude extract (12.5 ml/kg) of N. indicum seed gave 100% mortality of B. bengalensis. Humanness assessment study revealed that this plant orgin chemical caused low pain and sufferings to the target pests.

Toxicity of Nerium oleander leaf extract in mice.

Non-lethal dose of 70% ethanol extract of the Nerium oleander dry leaves (1000 mg/kg body weight) was subcutaneously injected into male and female mice once a week for 9 weeks (total 10 doses). One day after the last injection, final body weight gain (relative percentage to the initial body weight) had a tendency, in both males and females, towards depression suggesting a metabolic insult at other sites than those involved in myocardial function. Multiple exposure of the mice to the specified dose failed to express a significant influence on blood parameters (WBC, RBC, Hb, HCT, PLT) as well as myocardium. On the other hand, a lethal dose (4000 mg/kg body weight) was capable of inducing progressive changes in myocardial electrical activity ending up in cardiac arrest. The electrocardiogram abnormalities could be brought about by the expected Na+, K(+)-ATPase inhibition by the cardiac glycosides (cardenolides) content of the lethal dose.

Efecto del Nerium oleander L en modelo de corazón aislado de cobayo / Effect of Nerium oleander L in a guinea pig isolated heart

Se realizó un estudio farmacognóstico y farmacológico del Nerium oleander L. con el objetivo de determinar la presencia de metabolitos activos en la planta y corroborar en un modelo de corazón aislado de cobayo (Langerdorff) el efecto que como cardiotónico se le atribuye. Se encontró la presencia de glicósidos cardiotónicos y se produjo un aumento (p < 0,05) de la contractilidad del músculo cardíaco con el uso de un extracto fluido con menstruo hidroalcohólico al 30 porciento a dosis de 0,71 mg/g y 1,39 mg/g de peso del corazón, sin embargo, la dosis de 5,13 mg/g provocó arritmias. El flujo coronario no varió