Poisoning in ruminants caused by species of the genus Cestrum L. (Solanaceae) in Brazil: A review of toxicological and phytochemical evidence.

Some plant species of the genus Cestrum L. (Solanaceae family) are known to cause poisoning in farming animals in Brazil, negatively affecting the livestock sector. In this context, this study aimed to carry out a systematic review of the Cestrum species that cause poisoning in ruminants in Brazil and to list the main phytochemicals involved in these toxic activities that have already been identified. Scientific documents were retrieved in Google Scholar, PubMed®, ScienceDirect®, and SciELO databases. After applying the inclusion criteria, a total of 38 articles published between 1920 and 2023 were included in the present study. Cestrum axillare Vell. [Syn. Cestrum laevigatum Schltdl.], Cestrum corymbosum Schltdl., Cestrum intermedium Sendtn., and Cestrum parqui L'Hér. were found to have reported cases of poisoning in the Northeast, Southeast, and South of Brazil. Natural poisonings in ruminants caused by these species have been recorded in ten Brazilian states, mostly in Rio de Janeiro, Santa Catarina, Rio Grande do Sul, and Pernambuco. In general, Cestrum species cause liver damage and a clinical-pathological state characterized by acute liver failure of the poisoned animals. Cattle are more susceptible to poisoning by these plants, but there are reports of poisoning by C. axillare in goats and buffaloes as well. Several chemical constituents were identified in C. axillare and C. parqui, including some saponins and terpenoids that may be associated with the cases of poisoning. However, only one chemical compound has been identified in C. intermedium, and no phytochemical investigation has been carried out regarding toxic compounds in C. corymbosum. It is expected that future studies fill the gap in determining the toxic principles present in these species.

Essential Oil of Cestrum diurnum L.: GC/MS Analysis, in Vitro and in Silico Anti-HCoV-229E Effects and Inhibitory Activity against LPS-Induced Inflammation.

Cestrum diurnum L. (Solanaceae) is a fragrant ornamental tree cultivated in different parts around the world. In this study, the essential oil (EO) of the aerial parts was extracted by hydrodistillation (HD), steam distillation (SD) and microwave-assisted hydro-distillation (MAHD). GC/MS analysis of the three EOs revealed that phytol represents the major component in SD-EO and MAHD-EO (40.84 and 40.04 %, respectively); while in HD-EO it only represented 15.36 %. The SD-EO showed a strong antiviral activity against HCoV-229E with IC50 of 10.93 µg/mL, whereas, MAHD-EO and HD-EO showed a moderate activity with IC50 values of 119.9 and 148.2 µg/mL, respectively. The molecular docking of EO major components: phytol, octadecyl acetate and tricosane showed a strong binding to coronavirus 3-CL (pro). Moreover, the three EOs (50 µg/mL) decreased the levels of NO, IL-6 and TNF-α and suppressed IL-6 and TNF-α gene expression in LPS-induced inflammation model in RAW264.7 macrophage cell lines.

A Total of Eight Novel Steroidal Glycosides Based on Spirostan, Furostan, Pseudofurostan, and Cholestane from the Leaves of Cestrum newellii.

Previously, various steroidal glycosides were reported from plants of Cestrum species. However, phytochemical investigation has not been conducted on Cestrum newellii. A systematic phytochemical investigation of the leaves of C. newellii resulted in the isolation of eight novel steroidal glycosides (1-8), which were classified into three spirostanol glycosides (1-3), two furostanol glycosides (4 and 5), two pseudofurostanol glycosides (6 and 7), and one cholestane glycoside (8). In addition, three known cholestane glycosides (9-11) were isolated and identified. The structures of the new compounds were determined based on spectroscopic data and chemical transformations. Compounds 1 and 2 are spirostanol glycosides having hydroxy groups at C-2, C-3, C-12, and C-24 of the aglycone moiety. Although C. newellii is known to be a poisonous plant, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide assay exhibited that none of the isolated compounds were cytotoxic to HL-60 human promyelocytic leukemia cells.

In vitro anticancer and antioxidant potential of Cestrum species.

The methanolic extract of leaves and stem of Cestrum nocturnum and Cestrum diurnum were investigated for their antioxidant and anticancer attribute through standard methods. 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay was carried out to estimate the antioxidant activity of the extracts. Whereas, anticancer potential of extracts were tested against colon cancer cell line, HCT 116 and acute myeloid leukemia (AML) cell lines, THP-1. Results showed that extracts of both plants exhibited a very strong antioxidant activity in a dose dependent manner. In addition, both extracts efficiently increased the cell death in two different cancer cell lines. Moreover, DNA fragmentation analysis further strengthens the anticancer potential of extracts of both types of plants. Current study, therefore, provide a preliminary data highlighting the antioxidant and anticancer activities of methanolic extract of leaves and stem of Cestrum nocturnum and Cestrum diurnum.

Hypolipidemic effect of steroid compound from Cestrum diurnum (Solanaceae: Solanales) in normocholesterolemic albino rat.

The present study was carried out to establish the hypolipidemic effect of a phyto-steroid compound isolated from the chloroform: methanol extract of fresh mature leaves of the plant Cestrum diurnum (Solanaceae: Solanales). Change in the haematological parameters was studied in normocholesterolemic albino rats after oral administration of single dose of isolated phytosteroid (2 mg/ day) up to 15 days and compared with control rats. Application of phytosteroid fraction resulted in a significant reduction in total plasma cholesterol and free cholesterol levels. The plasma triglyceride levels also decreased significantly. A sharp increase in the HDL cholesterol level and a significant decrease in the LDL and VLDL amount were also documented. Free fatty acid level was significantly low in treated rats.

Comparação dos efeitos das folhas de Cestrum axillare Vell. com as saponinas isoladas em caprinos / Comparison of the effects of Cestrum axillare leaves with isolated saponins in goats

Cestrum axillare Vell. (formerly Cestrum laevigatum Schltd.), family Solanaceae, is the most important hepatotoxic plant in Brazil that causes acute poisoning. It occurs in the Southeast and Center-West regions and in coastal areas of the Northeast Brazil. Spontaneous poisoning was described in cattle, goats and sheep, with clinical signs evidenced within 24 hours after ingestion of the leaves and death within 48 hours after signs onset. The clinical signs observed in acute poisoning are apathy, anorexia, ruminal arrest, arched back, constipation with feces in small spheres, sometimes covered with mucus and blood streaks, muscle tremors, staggering gait and sometimes sialorrhoea. Neurological signs may be observed, due to interference in the urea cycle due to hepatic insufficiency resulting in hyperammonemia (hepatic encephalopathy). The main pathological finding is centrilobular hepatic necrosis. The toxic principle present in C. axillare was not yet definitively proven, but some authors attribute the toxicity of the plant to the presence of saponins gitogenin and digitogenin. However, it has not been determined whether the saponins present in C. axillare are responsible for the hepatotoxic effect of the plant. Thus, the objective of this work is to determine if the saponins are the compounds responsible for the hepatotoxic effects produced by the ingestion of the leaves of C axillare, using goats as experimental model. For this, the effects of the administration of the leaves were compared with those produced by the saponins isolated from the leaves in goats. Six goats were randomly assigned to three experimental groups that received [1] dry leaves of C. axillare (animals A1 and A2), [2] saponins extract from leaves (animals S1 and S2) or [3] control group (animals C1 and C2). For goats receiving the dry leaves the administered dose of plant was 10g/kg for one animal (A1) and 5g/kg for the other one (A2). For animals receiving the saponins extract, administration was done at a dose equivalent to 20g/kg repeated after 24 hours. The dry leaves administered at a dose of 10g/kg to a goat produced toxic effects, with alterations in biochemistry (indicating hepatic lesion) and histopathology showing centrilobular hepatic necrosis. At the dose of 5 g/kg of dry leaves, clinical signs of poisoning were not observed, but hepatic necrosis was found; after 15 days after the last administration, the hepatic parenchyma of this animal was already normal, with only hemorrhagic areas, demonstrating full regeneration. The administration of extracts of saponins containing gitogenin and digitogenin to goats did not produce significant toxic effects, proving that these compounds are not responsible for intoxication. In addition, goats are a good experimental model for studies of this intoxication.(AU)

Cestrum axillare Vell. (anteriormente C. laevigatum Schltd.), família Solanaceae, é a mais importante planta hepatotóxica do Brasil que causa intoxicação aguda. Tem ocorrência nas regiões Sudeste e Centro-Oeste e em áreas litorâneas do Nordeste. A intoxicação natural foi descrita em bovinos, caprinos e ovinos, com sinais clínicos evidenciados em até 24 horas após a ingestão das folhas e morte em até 48 horas após o início da sintomatologia. Os sinais clínicos observados na intoxicação aguda são apatia, anorexia, parada ruminal, dorso arqueado, constipação com fezes em formas de pequenas esferas, por vezes recobertas com muco e com estrias de sangue, tremores musculares, andar cambaleante e, às vezes, sialorreia. Podem ser observados sinais neurológicos, devido à interferência no ciclo da ureia pela insuficiência hepática resultando em hiperamonemia (encefalopatia hepática). O principal achado patológico é a necrose hepática centrolobular. O princípio tóxico presente no C. axillare ainda não está definitivamente comprovado, mas alguns autores atribuem a toxicidade da planta à presença das saponinas gitogenina e digitogenina. No entanto, ainda não foi determinado se as saponinas presentes em C. axillare são as responsáveis pelo efeito hepatotóxico da planta. Assim, o objetivo deste trabalho é determinar se as saponinas são os compostos responsáveis pelos efeitos hepatotóxicos produzidos pela ingestão das folhas de C. axillare, usando caprinos como modelo experimental. Para isto, foram comparados os efeitos da administração das folhas com os produzidos pelas saponinas isoladas destas folhas em caprinos. Foram utilizados seis caprinos, distribuídos aleatoriamente em três grupos experimentais que receberam [1] folhas secas de C. axillare (Caprinos A1 e A2), [2] extrato de saponinas das folhas (Caprinos S1 e S2), e [3] grupo controle (Caprinos C1 e C2). Para os caprinos que receberam as folhas secas a dose administrada de planta foi de 10g/kg para um animal (A1) e de 5g/kg para outro (A2). Para os animais que receberam o extrato de saponinas, a administração foi feita na dose equivalente a 20g/kg, repetida após 24 horas. Foi verificado que as folhas secas, quando administradas na dose de 10g/kg a um caprino, produziram efeitos tóxicos, com alterações na bioquímica (indicando lesão hepática) e histopatológica apresentando necrose hepática centrolobular. Na dose de 5g/kg de folhas secas, não foi observado sintomatologia clínica da intoxicação, mas houve necrose hepática; 15 dias após a última administração, o parênquima hepático deste animal já se encontrava normal, apenas com áreas hemorrágicas, demonstrando plena regeneração. A administração do extrato de saponinas contendo gitogenina e digitogenina a caprinos não produziu efeitos tóxicos significantes, comprovando não serem estes compostos os responsáveis pela intoxicação. Além disto, a espécie caprina é um bom modelo experimental para estudos desta intoxicação.(AU)

Spirostanol glucosides from the leaves of Cestrum laevigatum L.

Two new steroidal saponins, (25R)-spirost-5-ene-3ß,26ß-diol 3-O-α-L-rhamnopyranosyl-(1 → 4)-α-L-rhamnopyranosyl-(1 → 4)-[(1 → 2)-α-L-rhamnopyranosyl]-ß-D-glucopyranoside (1) and (25R)-spirost-6-ene-3ß,5ß-diol 3-O-α-L-rhamnopyranosyl-(1 → 4)-α-L-rhamnopyranosyl-(1 → 4)-[(1 → 2)-α-L-rhamnopyranosyl]-ß-D-glucopyranoside (2), along with the known diosgenin 3-O-α-L-rhamnopyranosyl-(1 → 4)-α-L-rhamnopyranosyl-(1 → 4)-ß-D-glucopyranoside (3), chonglouoside SL-5 (4) and Paris saponin Pb (5) were isolated from the leaves of Cestrum laevigatum. The structures of the compounds were determined using spectroscopic analyses including HRESI-MS, 1D and 2D NMR data, followed by comparison with data from the literature. Among them, two are particularly unique, compound 1 is the first (6)Δ-spirostanol saponin and compound 2 has an unusual C-26 hydroxyl in the (5)Δ-spirostanol skeleton. Antifungal testing showed a potent activity to formosanin C against Candida albicans and Candida parapsilosis. Evaluation of the cytotoxic activity indicated that compound 1 has a moderate activity against HL-60 and SF-295 cell lines, while compound 2 were active only against HL-60.

Antifungal Saponins from the Maya Medicinal Plant Cestrum schlechtendahlii G. Don (Solanaceae).

Bioassay-guided fractionation of the crude extract (80% EtOH) of the leaves of Cestrum schlechtendahlii, a plant used by Q'eqchi' Maya healers for treatment of athlete's foot, resulted in the isolation and identification of two spirostanol saponins (1 and 2). Structure elucidation by MS, 1D-NMR, and 2D-NMR spectroscopic methods identified them to be the known saponin (25R)-1ß,2α-dihydroxy-5α-spirostan-3-ß-yl-O-α-L-rhamnopyranosyl-(1 → 2)-ß-D-galactopyranoside (1) and new saponin (25R)-1ß,2α-dihydroxy-5α-spirostan-3-ß-yl-O-ß-D-galactopyranoside (2). While 2 showed little or no antifungal activity at the highest concentration tested, 1 inhibited growth of Saccharomyces cerevisiae (minimum inhibitory concentration (MIC) of 15-25 µM), Candida albicans, Cryptococcus neoformans, and Fusarium graminearum (MIC of 132-198 µM).

[Sperm immobilizing effect of leaves extracts of Cestrum parqui]. / Effet immobilisant des spermatozoïdes par les extraits des feuilles du Cestrum parqui.

OBJECTIVE: The leaves extracts of Cestrum parqui were reported to have spermicidal activity. The current investigation identified the spermicidal component of the extracts and evaluated its spermicidal potential in vitro, particularly the effects on sperm motility and vitality. METHODS: Sperms were prepared by discontinuous buoyant density gradient centrifugation and incubated with varying concentrations of extract from C. parqui (40-250 µg/ml) at 37°C and 5% CO(2). The mode of spermicidal action was evaluated by sperm motility and vitality at different intervals ranging from 5 to 240 minutes. Morphological changes in human spermatozoa after exposure to the extract were evaluated under transmission electron microscope. RESULTS: A dose- and time-dependent effect of this extract on sperm motility and viability was observed. The mean effective concentration of extracts that induced irreversible immobilization was 250 µg/ml. Transmission electron microscope revealed a significant damage to sperm membrane in head and acrosomal membranes, notable swelling and disruption. CONCLUSION: Our study indicates that this natural extract has potential spermicidal effect in vitro. It can adequately replace nonoxynol-9 in vaginal contraceptives to make them more vaginally safe and ecofriendly.

Bioefficacy of Plumbago zeylanica (Plumbaginaceae) and Cestrum nocturnum (Solanaceae) plant extracts against Aedes aegypti (Diptera: Culicide) and nontarget fish Poecilia reticulata.

In a search for natural products that could be used to control the vectors of tropical diseases, extracts of medicinal plants Plumbago zeylanica and Cestrum nocturnum have been tested for larvicidal activity against second, third, and fourth instar larvae of Aedes aegypti. The LC(50) values of all the extracts in different solvents of both the plants were less than 50 ppm (15.40-38.50 ppm) against all tested larval instars. Plant extracts also affected the life cycle of A. aegypti by inhibition of pupal development and adult emergence with increasing concentrations. The larvicidal stability of the extracts at five constant temperatures (19°C, 22°C, 25°C, 28°C, and 31°C) evaluated against fourth instar larvae revealed that toxicity of both plant extracts increases with increase in temperature. Toxicity studies carried out against fish species Poecilia reticulata, the most common nontarget organism in the habitats of A. aegypti, showed almost nil to meager toxicity at LC(50) and LC(90) doses of the plant extracts. The qualitative analysis of crude extracts of P. Zeylanica and C. nocturnum revealed the presence of bioactive phytochemicals with predominance of plumbagin in P. zeylanica and saponins in C. nocturnum. Partially purified plumbagin from P. zeylanica and saponins from C. nocturnum were obtained, and their presence was confirmed by thin-layer chromatography and biochemical tests. The bioassay experiment of partially purified secondary metabolites showed potent mosquito larvicidal activity against the fourth instar larval form. Therefore, this study explored the safer and effective potential of plant extracts against vector responsible for diseases of public health importance.

Steroidal saponins from the fruits of Cestrum ruizteranianum.

Seven spirostane and furostane-type glycosides were isolated from the aqueous methanolic extract of the fruits of Cestrum ruizteranianum and characterized mainly by 2D NMR spectroscopy and mass spectrometry. These known saponins belong to the delta5-spirostene and delta5-furostene series and are reported in this species for the first time.

Anti-inflammatory and analgesic components from "hierba santa," a traditional medicine in Peru.

"Hierba santa," a Peruvian herbal medicine, is used to alleviate many symptoms, including headache, hemorrhoids, fever, and rheumatism. Several Cestrum species are said to be the origin of hierba santa. Three lots of hierba santa: Cestrum auriculatum (herb 1 and herb 2) and C. hediundinum (herb 3), which were purchased from Peruvian markets at Cuzco (Andes area) and Equitos (Amazon area), respectively, were examined for their pharmacological activities and active components. Herbs 1-3 showed anti-inflammatory and analgesic activities in the in vivo writhing inhibition test in mouse and inhibited prostaglandin E(1)-, E(2)-, or ACh-induced contractions of guinea pig ileum in the Magnus method. Activity-based separation of each extract yielded cestrumines A and B, cestrusides A and B, a mixture of (+)- and (-)-pinoresinol glucosides, nicotiflorin, rutin, sinapoyl glucose, ursolic acid, beta-sitosteryl glucoside, and 2-sec-butyl-4,6-dihydroxyphenyl-beta-D: -glucopyranoside. Among them, cestrumine A and cestrusides A and B are new compounds. All three lots of hierba santa do not contain exactly the same active components.

Chemical characterization of new oxylipins from Cestrum parqui, and their effects on seed germination and early seedling growth.

Isolation, chemical characterization, and phytotoxicity of five new oxylipins, together with seven already known related compounds, from Cestrum parqui L' Hérl. is reported. All the structures were elucidated on the basis of their spectral data, especially 1D-(1H- and 13C-NMR, DEPT) and 2D-NMR (COSY, TOCSY, HSQC, HMBC, and NOESY). The configurations of the stereogenic C-atoms were determined by the Mosher's method. The compounds have been assayed for their phytotoxicity on Lactuca sativa at concentrations ranging between 10(-4) and 10(-8) M. The results of the phytotoxicity tests on the germination and growth of the test species, obtained by a cluster analysis, showed interesting relationship between the chemical structures of the compounds and their biological effects.

Cesdiurins I-III, steroidal saponins from Cestrum diurnum L.

Phytochemical investigation of a methanolic extract of leaves of Cestrum diurnum L (Solanaceae) resulted in isolation of several furostanol steroidal saponins, named cesdiurins I-III (1-3). Their structures of the isolated compounds were determined by spectroscopic analyses, including by use of 1D and 2D NMR spectroscopic techniques as well as by mass spectral analyses.

Laboratory evaluation of a phytosteroid compound of mature leaves of Day Jasmine (Solanaceae: Solanales) against larvae of Culex quinquefasciatus (Diptera: Culicidae) and nontarget organisms.

Crude mature leaves extract of Day Jasmine, Cestrum diurnum (Solanaceae: Solanales), was investigated for larvicidal activities against Culex quinquefasciatus, the vector of human filariasis. All the graded concentrations (0.1%, 0.2%, 0.5%, 1%, 1.5%, 2%, 2.5%, and 3% v/v) showed significant (P < 0.05) larval mortality, and results of regression equations revealed that the mortality rates were positively correlated with the concentrations of the extract (R (2) close to 1). LC(50), LC(95), and LC(99) values were calculated at different time intervals, and the lowest value was recorded at the 72-h bioassay for third-instar larvae. Significant changes in the larval mortality (F < 0.05) was also recorded between the periods of exposure and between instars during t test and single analysis of variance analysis. No mortality was noticed for nontarget organisms, such as Oreochromis niloticus niloticus and Chironomid larvae within 72 h of postexposure to 1%, 2%, and 3% crude plant extract under the laboratory condition. Qualitative and chromatographic analysis of the crude extract of C. diurnum revealed the presence of many bioactive phytochemicals. The bioassay experiment with the third-instar larval form established that the presence of a steroid compound (R (f), 0.67) was responsible for mosquitocidal activity.

[Experimental study on antitumor effect of extracts from Cestrum nocturnum in vivo].

OBJECTIVE: To observe the antitumor effects of extracts from Cestrum nocturnum (CN) in vivo. METHODS: The S180-mice model was used to observe the tumor-inhibition rate of CN and the H22-mice model was used to test the survival time. RESULTS: The experiment in S180-mice demonstrated that the n-butanol and polysaccharides extracts from CN had obvious effects on tumor inhibition. Its inhibitory rates were 52.30%, 46.75%, 42.28%, 43.19%, 37.96% and 31.82% respectively in the mice administrated the n-butanol and polysaccharides extracts from CN with 30 mg/kg, 20 mg/kg and 15 mg/kg weight dosage. It was noted that tumor formation postponed in mice treated with the n-butanol and polysaccharides extracts from CN compared with the control panel and tumor growth became slower; The n-butanol and polysaccharides extracts from CN could also greatly enhance the life span of mice with H22 ascitic tumors by 116.43%, 44.52%, 20.54%, 109.52%, 112.61% and 115.01%, respectively. The inhibit effects of n-butanol fraction extracts from CN had direct relationship with dose, while the polysaccharides fraction extracts from CN had no obvious relationship with dose. CONCLUSION: The n-butanol and polysaccharides extracts of CN are able to inhibit tumor growth and prolong the lifetime of the tumor-bearing mice in a dose-dependent pattern.

Purification of a natural insecticidal substance from Cestrum parqui (Solanaceae).

Cestrum parqui is a shrub originating from Chile used in Tunisia as an ornamental plant. The toxicity of this plant was observed for the first time on the Desert Locust Schistocerca gregaria. It was shown that the toxicity of the plant is concentrated in the Crude Saponic Extract (CSE). In the present research we try to study chemically this fraction to isolate the active substances. The CSE was separated by column chromatography and the obtained fractions were biologically tested on Schistocerca larvae. We observed that only the fraction F9 was active, the separation of this fraction on preparative plates permits to isolate the active compound called S1. The tentative of structure elucidation of the natural substance S1 using 1H and 13C NMR shows that S1 is an heterosidic saponin. Structural modification of S1 structure provoke its activity lose.

Insecticidal effects of various concentrations of selected extractions of Cestrum parqui on adult and immature Ceratitis capitata.

Aqueous extracts of Cestrum parqui L'Héritier (Solanaceae) were evaluated at different concentrations in several stages of Ceratitis capitata (Wiedemann) (Diptera: Tephritidae). For adults, the study was extended to extracts obtained with several solvents of an increasing degree of polarity. Aqueous extracts from C. parqui showed a high toxicity to neonate larvae when ingested through diet, inhibiting pupation at a concentration above 0.6%. Lower concentrations delayed the larval development and reduced the percentages of pupae formed and adult emergence. An LC50 = 0.9% after 3 d of continuous ingestion of C. parqui aqueous extracts could be calculated, whereas extracts obtained with organic compounds were nearly innocuous except with the use of the solvent methanol/water (80:20), the more polar of those tested, that killed 12.5% of adults. Aqueous extracts were also harmful to adults by diminishing the reproductive potential, which implies a significant effect on the offspring. Egg contact with insecticide or dipping third instars did not cause any adverse effect, supporting the hypothesis that only by means of ingestion can the toxic compounds of C. parqui reach the target. Our results showed that C. parqui components causing C. capitata mortality are mostly dissolved in water and not in organic solvents, which point out their polar character.

Biocontrol efficacy of Cestrum diurnum L. (Solanaceae: Solanales) against the larval forms of Anopheles stephensi.

Cestrum diurnum L. (Solanaceae: Solanales) is a multistemmed shrub that is also known as 'Day Jasmine'. The efficacy of the plant as a strong biocontrol agent of the larval anopheline mosquito was evaluated in the laboratory and bioactive ingredients were characterized by thin layer chromatography (TLC) and infrared (IR) analysis. Thin layer chromatographic analysis revealed the presence of steroidal compound as a bioactive ingredient and IR analysis reported the presence of O-H group, C-S bond as a -CH2S group, H bonded -C=O group and carbonyl stress of esters. The LC50 value of the active ingredient was determined as 0.70, 0.89, 0.90 and 1.03 mg/100 mL-1, for 1st, 2nd, 3rd and 4th instar larva respectively in 24 h study period. The active component was analyzed by GLC and GLC-MS and altogether 14 components were detected.

Amelioration of adjuvant-induced arthritis by ursolic acid through altered Th1/Th2 cytokine production.

The objective of the study was to investigate the activity of ursolic acid (UA) on proinflammatory (Th1) and anti-inflammatory (Th2) cytokines in the peripheral blood of arthritic balb/c mice. Ursolic acid is ubiquitous in the plant kingdom and is a constituent of numerous plants which are having diversified phylogenetic origin and taxonomic position. We applied Cytometric bead array (CBA) technology for simultaneously measurement of these cytokines in adjuvant inflammatory arthritis induced mice treated with ursolic acid in graded oral doses. Cytometric bead array uses the sensitivity of amplified fluorescence detection by flowcytometer to measure soluble analytes in a particle based immune assay. This assay can accurately quantitate five cytokines in a 50 microl sample volume. The T-helper (Th1) deviated cells produce detectable level of tumor necrosis factor (TNF-alpha), interleukin-2 (IL-2) and interferon-gamma (IFN-gamma), while the Th2 deviated cells produce significant amount of interleukin-4 (IL-4) and interleukin-5 (IL-5). Oral administration of UA at doses of 10, 20, 40, 80 and 160 mg kg(-1) per oral dose inhibited the presence of IL-2, IFN-gamma and TNF-alpha in the peripheral blood.