Protective effect of zinc oxide nanoparticles synthesized using Cassia alata for DSS-induced ulcerative colitis in mice model.

The most prevalent form of inflammatory bowel disease (IBD), ulcerative colitis (UC), is characterized by persistent inflammation of the colorectal mucosa. It is asymptomatic, whereas Crohn's disease (CD) causes patchy lesions in the gastrointestinal tract. Men and women suffer equally from ulcerative colitis, which usually strikes in the second and third decades of life and becomes more common in senior citizens. In the present study, we produced zinc oxide nanoparticles using the natural herbal plant, Cassia alata. Zinc oxide nanoparticles have remarkable antimicrobial and antitumor benefits in the field of biomedical science. Furthermore, the synthesized zinc oxide nanoparticles (ZnO NPs) were characterized using UV, XRD, FTIR, and SEM analyses. The XRD analysis confirmed the crystallite nature and purity of the synthesized nanoparticles. Zinc oxide nanoparticles with a uniform size and partially agglomerated morphology were verified by SEM analysis. We investigated the protective effects of environmentally friendly zinc oxide nanoparticles in dextran sodium sulfate-induced ulcerative colitis mouse models. Green synthesized Cassia alata zinc oxide nanoparticles (CA ZnO NPs) reversed weight loss, disease activity index, colon shortening, and colon histological damage. Zinc oxide nanoparticles reduce hypersensitivity, oxidative stress, and inflammation, and protect the mucosal layer. Green synthesized CA ZnO NPs demonstrated protection against dextran sodium sulfate-induced ulcerative colitis via anti-inflammatory activity.

Cassia alata's dual role in modulating MUC2 expression in Eimeria papillata-infected jejunum and assessing its anti-inflammatory effects.

Coccidiosis poses significant hazards to animals, particularly in terms of compromised health, reduced productivity, and economic losses in livestock farming. The conventional treatments for coccidiosis often involve synthetic drugs, contributing to concerns about drug resistance and environmental impact. The pressing need for eco-friendly alternatives is highlighted in this study, emphasizing the importance of exploring medicinal plants like Cassia alata leaf extracts (CAE) against Eimeria papillata-induced infection in mice. The CAE exhibited significant phenolic (2.17 ± 0.03 g/100 g) and flavonoid (0.14 ± 0.01 g/100 g) content and demonstrated notable antioxidant activity. In infected mice, the CAE treatment led to a substantial reduction in oocyst output (~6 fold), ameliorating necrotic enteritis and inflammatory changes in the jejunum. Additionally, CAE treatment increased goblet cell numbers (9.3 ± 0.1 / villus) and decreased macrophage infiltration in the intestinal villi. Molecular analyses revealed CAE's positive modulation of MUC2 gene and notably reduced the levels of pro-inflammatory cytokines (specifically IL-1ß, IL-10, and IFN-γ) when contrasted with the infected cohort. Furthermore, CAE treatment significantly reduced nitric oxide levels (44.03 ± 2.4 µmol/mg), showcasing its anti-inflammatory properties. The findings of this study not only contribute to the understanding of CAE's therapeutic potential but also underscore the importance of seeking eco-friendly alternatives in the face of coccidiosis challenges, addressing both the well-being of animals and the sustainability of agricultural practices. RESEARCH HIGHLIGHTS: Cassia alata extract (CAE) exhibited significant phenolic and flavonoid content, displaying notable antioxidant activity. In infected mice, CAE treatment led to a substantial reduction in oocyst output, ameliorating necrotic enteritis and inflammatory changes in the jejunum. CAE treatment increased goblet cell numbers and decreased macrophage infiltration in the intestinal villi, while molecular analyses revealed its positive modulation of the MUC2 gene and notable reduction in pro-inflammatory cytokine levels. Additionally, CAE treatment significantly reduced nitric oxide levels, showcasing its anti-inflammatory properties.

In vitro antimicrobial efficacy of Cassia alata (Linn.) leaves, stem, and root extracts against cellulitis causative agent Staphylococcus aureus.

BACKGROUND: Cellulitis is a common skin disease encountered in medical emergencies in hospitals. It can be treated using a combination of antibiotics therapy; however, the causative agent Staphylococcus aureus has been reported to develop resistance towards the currently used antibiotics. Therefore, the search for more alternative herbal origin antimicrobial agents is critical. AIM: In this study, maceration and Soxhlet extraction of the whole plant of Cassia alata Linn. (leaves, roots, and stem) were performed using four solvents with different polarities, namely n-hexane, ethyl acetate, ethanol and distilled water. The crude extracts were screened using agar well diffusion, colorimetric broth microdilution, grid culture and bacterial growth curve analysis against Staphylococcus aureus. The phytochemicals in the crude extracts were identified using Gas Chromatography-Mass Spectrometry (GC-MS). RESULTS: Agar-well diffusion analysis revealed that extraction using ethyl acetate showed the largest inhibition zone with an average diameter of 15.30 mm (root Soxhlet extract) followed by 14.70 mm (leaf Soxhlet extract) and 13.70 mm (root maceration extract). The lowest minimum inhibitory and minimum bactericidal concentration in root Soxhlet extract using ethyl acetate was 0.313 and 0.625 µg µL-1, respectively. Our study proved that crude extract of the plant suppressed the growth of S. aureus as evidenced from a significant regression extension (p < 0.06, p = 0.00003) of lag phase for 6 h after the treatment with increased concentration. Based on the GC-MS analysis, 88 phytochemicals consist of fatty acids, esters, alkanes, phenols, fatty alcohols, sesquiterpenoids and macrocycle that possibly contributed to the antimicrobial properties were identified, 32 of which were previously characterized for their antimicrobial, antioxidant, and anti-inflammatory activities. CONCLUSION: Ethyl acetate crude extract was better than the other investigated solvents. The root and stem of C. alata showed significant antimicrobial efficacy against S. aureus in this study. The remaining 56 out of 88 phytochemicals of the plant should be intensively studied for more medicinal uses.

The complete chloroplast genome of Senna alata (L.) Roxb., an important medicinal plant from the Philippines.

Senna alata, a flowering shrub, is widely cultivated in the Philippines for its anti-fungal properties. Despite this, its chloroplast genome is not yet established. We assembled and annotated the complete chloroplast genome of accession from the germplasm collection of the Institute of Crop Science, University of the Philippines, Los Baños, using Illumina sequencing data. The complete cp genome was 159,176-bp long characterized by a large single copy of 88,769 bp, short single-copy of 18,301 bp and a pair of inverted repeat regions of 26,053 bp each. The overall GC content of the chloroplast genome was 36.4%. The plastome comprised 37 tRNA genes, 8 rRNA genes and 78 mRNA genes. Phylogenetic analysis showed that S. alata is closely related to S. siamea.

Effects of Cellulase and Lactiplantibacillus plantarum on the Fermentation Parameters, Nutrients, and Bacterial Community in Cassia alata Silage.

Silage Cassia alata (CA) can alleviate feed shortage in some areas to a certain extent and reduce feed costs. The present research evaluated the effect of cellulase (CE) and Lactiplantibacillus plantarum (LP) on the fermentation parameters, nutrients, and bacterial community of CA silage. Chopped CA was ensiled with three different treatments, namely, no inoculant (CK), CE, and LP, and the indexes were determined on the 2nd, 6th, 14th, and 30th days of silage fermentation. The fermentation parameters indicate that the pH value of the three groups decreased and then increased with the ensilage process, and the lowest value was observed on the 14th day. The CK and LP groups attained the highest value on the 30th day, while the CE group attained the highest value on the 2nd day. Additionally, the pH value and NH3-N content were significantly lower (P < 0.05) in the CE and LP groups than in the CK group. In terms of nutrients, crude protein (CP) contents significantly increased (P < 0.05) in the CE and LP groups on the 30th day. The neutral detergent fiber (NDF) and acid detergent fiber (ADF) contents of the CE group were significantly and negatively associated with fermentation time, and the water-soluble carbohydrate (WSC) contents of the three groups were significantly lower during ensiling. In comparison with the CK group, the NDF and ADF contents were significantly reduced (P < 0.05), and the WSC content increased (P < 0.05) in the CE group on day 30. Sequencing analysis of bacterial communities showed that Lactobacillus became the most dominant genus in the ensilage process. Moreover, both CE and LP groups increased the abundance of Lactobacillus and decreased that of Klebsiella, Weissella, and Acetobacter in comparison to the CK group, in which LP had a better effect. CE and LP could further improve the silage quality of CA, and LP had a more significant effect in reconstructing the bacterial community in the silage environment.

Indole Alkaloids and Chromones from the Stem Bark of Cassia alata and Their Antiviral Activities.

The Cassia (Leguminosae) genus has attracted a lot of attention as a prolific source of alkaloids and chromones with diverse structures and biological properties. The aim of this study is to screen the antiviral compounds from Cassia alata. The extract of the stem bark of this plant was separated using silica gel, MCI, ODS C18, and Sephadex LH-20 column chromatography, as well as semi-preparative HPLC. As a result, three new indole alkaloids, alataindoleins A-C (1-3); one new chromone, alatachromone A (4); and a new dimeric chromone-indole alkaloid, alataindolein D (5) were isolated. Their structures were determined by means of HRESIMS and extensive 1D and 2D NMR spectroscopic studies. Interestingly, alataindolein D (5) represents a new type of dimeric alkaloid with an unusual N-2-C-16' linkage, which is biogenetically derived from a chromone and an indole alkaloid via an intermolecular nucleophilic substitution reaction. Compounds 1-5 were tested for their anti-tobacco mosaic virus (TMV) and anti-rotavirus activities, and the results showed that compounds 2-4 showed high anti-TMV activities with inhibition rates of 44.4%, 66.5%, and 52.3%, respectively. These rates were higher than those of the positive control (with inhibition rate of 32.8%). Compounds 1 and 5 also showed potential anti-TMV activities with inhibition rates of 26.5% and 31.8%, respectively. In addition, compounds 1-5 exhibited potential anti-rotavirus activities with therapeutic index (TI) values in the range of 9.75~15.3. The successful isolation and structure identification of the above new compounds provided materials for the screening of antivirus drugs, and contributed to the development and utilization of C. alata.

Cyclotrisiloxan and ß-Sitosterol rich Cassia alata (L.) flower inhibit HT-115 human colon cancer cell growth via mitochondrial dependent apoptotic stimulation.

Cancer traits dependent chemo and radiotherapy display acute toxicity and long-term side effects. Since last two decades, researchers investigated a new anticancer agents derived from plants. Cassia alata (L.) is a medicinal herb distributed in the tropical and humid regions. In this study, C. alata flower methanol extract (CME) have been prepared using cold percolation method and the phytochemical components were identified using GC-MS analysis. CME have been used to study the antiproliferative and apoptosis properties against human colon cancer HT-115 colon cancer cells, its molecular mechanism have been explored. 0.2 mg/mL dose of CME, inhibited 50% of HT-115 colon cancer cell growth after 48hr was confirmed the significant antiproliferation effect. In normal cells such as Vero cells and hMSCs, 0.2 mg/mL dose of CME shown only 4% and 5% growth inhibition confirmed the HT-115 cell specific cytotoxic effect. This effect might be due to the availability of phytoactive biomolecules in CME such as, cyclotrisiloxan, beta-sitosterol and alpha-tocopherol have been confirmed by GC-MS. Most interestingly, PI and AO/ErBr staining of CME treated HT-115 cells shown early (25%), pro (17%) and late (8%) apoptotic and 3% necrotic cells after 48 hr. Treatment with CME extract showed potential effect on the inhibition of protumorigenic inflammatory and oxidative stress genes. Protumorigenic COX-2/PGE-2 and TNF-α/NF-κB immune axis were normalized after CME treatment. Amounts of both apoptosis related mRNA p53, Bax, caspase 3 and p21 genes were upregulated, whereas it resulted in significant reduction in the anti-apoptotic marker mdm2 and Bcl-2 genes. In conclusion, bioactive compounds present in CME potentially inhibit HT-115 colon cancer cell proliferation via an inhibition of protumorigenic immune axis and stimulation of mitochondria dependent apoptotic pathway without necrotic effect.

A new, innovative, and safe treatment in vitiligo: Results of a randomized, double-blinded, parallel-group study.

Vitiligo is a chronic autoimmune disease affecting around 1% of the population worldwide. No existing treatment is giving fully satisfactory results. Further investigations are welcomed for innovative and safe treatments bringing better results. This trial aimed to compare the efficacy and tolerance of various treatment protocols on vitiligo lesions. Four randomized groups of 10 patients with vitiligo covering 8% to 14% of skin surface, except hands and feet were assigned during 8 weeks to (a) UVB microphototherapy 300 to 320 nm (Bioskin-) 1 x week; (b) VITILSI- gel 2 x day; (c) VITILSI- gel 2xday + Bioskin- 1 x week; and (d) placebo 2 x day. Efficacy of the treatment was assessed by planimetry, comparing the photographs of the patients taken at baseline and after 8-week treatment. After completion of the treatment, the increase of the pigment area was 28% in G1 (Bioskin-), 19% in G2 (VITILSI-), 41% in G3 (Bioskin- + VITILSI-) and null in G4. No subject stopped the treatment and no side effect was observed. It was demonstrated that the gel under study was able per se to induce repigmentation in vitiligo lesions and that the results were significantly better when combined with NB-UVB. The protocols used in this trial resulted safe and efficient.

Alatains A and B, unique hetero-dimeric polyphenols from Cassia alata and their anti-tobacco mosaic virus activity.

Two structurally unique polyphenols, alatains A (1) and B (2), were isolated from the bark of Cassia alata. Their structures were elucidated on the basis of spectroscopic analysis. Compounds 1 and 2 represent a new type of hetero-dimeric polyphenols with a C-14-C-5' linkage, biogenetically formed by an unusual intermolecular oxidative phenol-coupling reaction between a chromone unit and an isocoumarin moiety. Moreover, compounds 1 and 2 showed significant anti-tobacco mosaic virus (anti-TMV) inhibition IC50 values of 18.8 and 11.4 µM, respectively. Alatains A and B also exhibited promising protective effects on TMV infection of the host plants (Nicotiana tabacum) with the inhibition rates of 54.6% and 69.7% at the concentration of 20 µM, respectively. The results provided a new structural template for potential anti-TMV agent discovery.

Chemical constituents, usage and pharmacological activity of Cassia alata.

Cassia alata or locally known as Ketepeng Cina (Indonesia) and Gelenggang (Malaysia) has been used as a traditional medicine to treat various diseases, especially skin diseases. In addition, C. alata has been reported to have potential anti allergic, anti inflammatory, antioxidant, anticancer, antidiabetic, and antifungal. Metabolite compounds that have been isolated from C. alata include flavones, flavonols, flavonoids glycosides, alatinon, alanonal and ß-sitosterol-ß-D-glucoside. The compounds have been isolated mainly from the leaves. Further identification is needed to discover the secondary metabolites from other parts of the plant such as seed, flower and bark which are reported to have potent antibacterial and antifungal activity. Therefore, this article highlights the secondary metabolites and biological activity of this plant which has been shown to have pharmacological properties against selected diseases.

Effects of Cassia alata Linn on bowel function recovery following surgery for gynecological cancer: A randomized controlled trial.

BACKGROUND: Postoperative ileus is a common complication following abdominal surgery. This study was undertaken to determine the effectiveness and safety of Cassia alata Linn in stimulating gastrointestinal function recovery among women with gynecologic cancer who had undergone laparotomy for surgical staging or cytoreductive surgery. METHODS: A total of 90 participants were randomly allocated to postoperative consumption of either Cassia alata Linn tea (n = 45) or warm water (N = 45). Outcomes of interest included time to first passage of flatus, time to first defecation, time to toleration of solid food, and ileus symptoms. Differences between the groups were measured as mean difference (MD) or odds ratio (OR) with 95% confidence interval (CI). RESULTS: Women allocated to the Cassia alata Linn group had shorter times to first passage of flatus (MD -8.53; 95% CI -3.69, -13.38 h) and first defecation (MD -19.83; 95% CI -11.18, -28.48 h) compared with controls. There were no differences in terms of time to toleration of solid food (MD 0.26; 95% CI -2.13, 1.61 h) or ileus symptoms (OR 0.10; 95% CI 0.01, 1.94) between the two groups. CONCLUSION: Postoperative consumption of Cassia alata Linn hastens gastrointestinal function recovery after laparotomy in women with gynecologic cancer.

Characterisation of the Convective Hot-Air Drying and Vacuum Microwave Drying of Cassia alata: Antioxidant Activity, Essential Oil Volatile Composition and Quality Studies.

The preservation of active constituents in Cassia alata through the removal of moisture is crucial in producing a final product with high antioxidant activity. This study aims to determine the influences of various drying methods and drying conditions on the antioxidant activity, volatiles and phytosterols content of C. alata. The drying methods used were convective drying (CD) at 40 °C, 50 °C and 60 °C; freeze drying; vacuum microwave drying (VMD) at 6, 9 and 12 W/g; and two-stage convective pre-drying followed by vacuum microwave finish drying (CPD-VMFD) at 50 °C and 9 W/g. The drying kinetics of C. alata are best described by the thin-layer model (modified Page model). The highest antioxidant activity, TPC and volatile concentration were achieved with CD at 40 °C. GC-MS analysis identified the presence of 51 volatiles, which were mostly present in all samples but with quantitative variation. The dominant volatiles in fresh C. alata are 2-hexenal (60.28 mg 100 g-1 db), 1-hexanol (18.70 mg 100 g-1 db) and salicylic acid (15.05 mg 100 g-1 db). The concentration of phytosterols in fresh sample was 3647.48 mg 100 g-1 db, and the major phytosterols present in fresh and dried samples were ß-sitosterol (1162.24 mg 100 g-1 db). CPD-VMFD was effective in ensuring the preservation of higher phytosterol content in comparison with CD at 50 °C. The final recommendation of a suitable drying method to dehydrate C. alata leaves is CD at 40 °C.

Potential of Cassia alata L. Coupled with Biochar for Heavy Metal Stabilization in Multi-Metal Mine Tailings.

To explore the effect of different biochars on Cassia alata L. growth and heavy metal immobilization in multi-metal mine tailings, a 100-day pot experiment was conducted. Three biochars derived from Hibiscus cannabinus core (HB), sewage sludge (SB) and chicken manure (MB), were added to mine tailings at rates of 0.4%, 1% and 3% (w/w). The results showed that the root biomass, shoot biomass, plant height and root length were 1.2-2.8, 1.7-3.2, 1-1.5 and 1.6-3.3 times of those in the control group, respectively. Pb, Zn, Cu, Cd and As contents in the shoot decreased by 63.9-89.5%, 46.9-66.0%, 32.7-62.4%, 40.4-76.4% and 54.9-77.5%, respectively. The biochar significantly increased the pH and decreased the mild acid-soluble Pb and Cu concentrations in the mine tailings. Specifically, SB immobilized Pb and Cu better than MB and HB did, although it did not immobilize As, Zn or Cd. Meanwhile, more attention should be paid to the potential As release as the biochar application rate increases. In conclusion, Cassia alata L. coupled with 3% of SB could be an effective measure for restoring multi-metal mine tailings. This study herein provided a promising ecological restoration technique for future practice of heavy metal stabilization in mine tailings.

Cellulose nanocomposite films with in situ generated silver nanoparticles using Cassia alata leaf extract as a reducing agent.

Cotton linters were dissolved in aq. (8% LiOH+15% urea) that was pre-cooled to -12.5°C. Using this solution cellulose gel films were prepared by regeneration method with ethyl alcohol as a coagulant. These wet films were diffused with 10wt% Cassia alata leaf extract that acted as a reducing agent. The leaf extract diffused cellulose wet films were used as the matrix. The wet matrix films were dipped individually in lower concentrated 1-5mM aq.AgNO3 source solutions in the presence of sunlight and allowed the solutions to react with the diffused leaf extract reducing agent which in situ generated the silver nanoparticles (AgNPs) inside the films as well as in the source solution. The AgNPs formed in the source solution were observed by transmission electron microscope (TEM) and scanning electron microscope (SEM) while those formed in situ the films were observed by SEM and the particle size distribution was determined. The cellulose/AgNP composite films showed good antibacterial activity against Escherichia coli bacteria. These nanocomposite films were also characterized by Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), thermogravimetric analysis (TGA) and tensile tests. At temperatures below 300°C, the thermal stability of the nanocomposite films was lower than that of the matrix due to the catalytic effect of AgNPs. The nanocomposite films also possessed good tensile properties. The ecofriendly cellulose/AgNP composite films with good antibacterial activity and tensile properties can be considered for medical applications like dressing materials.

Evaluation of the Antiplasmodial Activity and Lethality of the Leaf Extract of Cassia alata L. (Fabaceae).

OBJECTIVE: Cassia alata L. (Fabaceae), one of the three plants contained in Saye, a polyherbal antimalarial remedy was assessed for its antimalarial potential and safety in mice. METHODOLOGY: Organic extracts were prepared from the leaves and tested on the D 10 chloroquine-sensitive strain of Plasmodium falciparum using the parasite lactate dehydrogenase assay. The 4 days suppressive test using Plasmodium berghei in mice was used to evaluate the in vivo antiplasmodial activity of the extracts. Animals were treated by oral route, once a day with 50, 100, 250 and 400 mg kg -1 b.wt., of the extracts. The acute toxicity of the extracts was assessed in mice according to Thompson and Weil method. The lethal effects of the extracts on animal's body weight, tissues, biochemical and haematological parameters were determined at 823.5, 1235.5, 1853 and 2779.5 mg kg -1 b.wt., respectively. RESULTS: The dichloromethane/methane (1:1, v/v) extract of Cassia alata was the most active against Plasmodium falciparum. The mean percent suppression of parasitemia in mice was equal to 22.5, 41.8 and 45.2% at 50, 250 and 400 mg kg -1 b.wt., respectively. No death and no clinically significant changes were recorded in mice. The maximum non-lethal dose was more than 16875 mg kg -1 in animals. No significant changes were observed in body weight, tissues morphology, biochemical and hematological parameters at doses above or equal to 2779.5 mg kg -1 b.wt. CONCLUSION: The dichloromethane/methanol leaf extract of Cassia alata had a good to moderate in vitro and in vivo antiplasmodial activity and was found to have low toxicity at high doses in tested animals.

[A new furan-2-carboxylic acid from stem bark of Cassia alata].

A new furan-2-carboxylic acid, 5-[3-(hydroxymethyl)-4,5-dimethoxyphenyl]-3-methylfuran-2-carboxylic acid(1),has been isolated from the bark of Cassia alata by using various chromatographic techniques. It displayed cytotoxicity against NB4, A549, SHSY5Y, PC3 and MCF7 cell lines with IC50 values of 2.5, 1.2, 2.2, 3.6 and 1.9 µmol•L⁻¹, respectively.

Senna alata leaves are a good source of propelargonidins.

Proanthocyanidins (PA) in Senna alata leaves were investigated by thiolysis with benzyl mercaptan, LC-MS and NMR and consisted of almost pure propelargonidins with <6% procyanidins, had B-type linkages and a mean degree of polymerisation of three. Epiafzelechin was the major flavan-3-ol subunit (>94%) and epicatechin a minor constituent (6.4%) in residual PA and mainly detected as an extension unit.

Assessment of antibacterial activity of crude leaf and root extracts of Cassia alata against Neisseria gonorrhea.

BACKGROUND: Gonorrhea is a major sexually transmitted disease worldwide and for its control, effective treatment is essential. However as more strains of Neisseria gonorrhoeae continuously develop resistance to several drugs, this strategy obliges scientists to discover newer effective drugs. OBJECTIVES: To ascertain whether crude leaf and root extracts of Cassia alata (Caesalpiniaceae) have antimicrobial activity against clinically resistant Neisseria gonorrhoeae bacteria. To determine and compare the MICs of their ether and methanol extracts. MATERIALS AND METHODS: Ether and methanol extracts were prepared from the plant parts. 12-375mg/ml of serially diluted ether extracts in DMSO and methanol extracts in water were tested using agar-well diffusion method against Neisseria gonorrhea clinical isolate cultured on MTM agar. MICs were determined from corresponding concentration-response curves. Ceftriaxone was used as positive control, whereas DMSO and water as negative controls. RESULTS: All the crude extracts showed concentration-dependent Neisseria gonorrhea inhibition. Ether extracts for both leaves and roots gave lower MICs compared to those of methanol. Ether root extract showed the highest potency. CONCLUSIONS: Both the leaf and the root of Cassia alata plant have activity against clinically resistant Neisseria gonorrhoeae; the root having the higher activity. Lipophilic solvent, ether, give more potent antigonorrhoeal extracts. As expected Cassia alata plant in Central Uganda also has antibacterial activity.

Astragalin from Cassia alata induces DNA adducts in vitro and repairable DNA damage in the yeast Saccharomyces cerevisiae.

Reverse phase-solid phase extraction from Cassia alata leaves (CaRP) was used to obtain a refined extract. Higher than wild-type sensitivity to CaRP was exhibited by 16 haploid Saccharomyces cerevisiae mutants with defects in DNA repair and membrane transport. CaRP had a strong DPPH free radical scavenging activity with an IC(50) value of 2.27 µg mL(-1) and showed no pro-oxidant activity in yeast. CaRP compounds were separated by HPLC and the three major components were shown to bind to DNA in vitro. The major HPLC peak was identified as kampferol-3-O-ß-d-glucoside (astragalin), which showed high affinity to DNA as seen by HPLC-UV measurement after using centrifugal ultrafiltration of astragalin-DNA mixtures. Astragalin-DNA interaction was further studied by spectroscopic methods and its interaction with DNA was evaluated using solid-state FTIR. These and computational (in silico) docking studies revealed that astragalin-DNA binding occurs through interaction with G-C base pairs, possibly by intercalation stabilized by H-bond formation.

Cassia alata leaf extract induces cytotoxicity in A549 lung cancer cells via a mechanism that is caspase 8 dependent / El extracto de la hoja de cassia alata induce citotoxicidad en las células A549 del cáncer pulmonar humano a través de un mecanismo dependiente de la caspasa 8

OBJECTIVE: To evaluate the cytotoxic effect of a hexane extract of Cassia alata leaves in A549 lung cancer cells. METHOD: Parental A549 lung cancer cells were exposed to various concentrations (100"180 µg/ml) of Cassia alata leaf extract for 24 hours. Following treatment, the cells were evaluated using the 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay to determine the cytotoxic effect of the extract. Caspase 8, 3 and 9 negative A549 cells were also prepared using lentiviral based shRNA knockdown of the caspase 8, 3 and 9 genes, respectively. The cytotoxic effect of Cassia alata leaf extract was then evaluated in these knockdown cells using the MTT assay. Chemical analysis was performed on the extract using high performance liquid chromatography (HPLC). RESULTS: Cassia alata extract was cytotoxic in parental and caspase-9 negative, but not caspase 3 and 8 negative A549 cells. The IC50 values were 143 µg/ml and 145 µg/ml in parental and caspase 9 negative A549 cells respectively. The flavanoid kaempferol was identified as a constituent of Cassia alata leaf extract. CONCLUSIONS: Cassia alata produces cytotoxicity in A549 cancer cells that is mediated by caspase 8 activation. This effect may be attributable to kaempferol.

OBJETIVO: Evaluar el efecto citotóxico de un extracto de hexano de hojas de Cassia alata en las células A549 del cáncer pulmonar. MÉTODO: Células A549 parentales del cáncer pulmonar fueron expuestas a varias concentraciones (100-180 µg/ml) de un extracto de la hoja de Cassia alatadurante 24 horas. Tras el tratamiento, las células fueron evaluadas usando el ensayo de bromuro de 3-(4,5-dimetiltiazol-2-il)-2,5-difeniltetrazolio (MTT) a fin de determinar el efecto citotóxico del extracto. También se prepararon células A549 negativas caspasa 8, 3 y 9 mediante silenciamiento génico vía ARN (shRNA knockdown) de los genes de las caspasas 8, 3 y 9 respectivamente, sobre la base de la inserción de vectores lentivirales. Entonces, usando un ensayo MTT se procedió a evaluar el efecto citotóxico del extracto de hojas de Cassia alataen éstas células genéticamente modificadas. Se realizó un análisis químico del extracto utilizando cromatografía líquida de alta eficacia. (HPLC). RESULTADOS: El extracto de Cassia alata resultó ser citotóxico en las células A549 negativas parentales y caspasa 9, pero no en las negativas caspasa 3 y 8. Los valores de IC50 fueron 143 µg/ml y 145 µg/ml en las células A549 negativas parentales y caspasa 9 respectivamente. El flavonol kaempferol fue identificado como un constituyente del extracto de las hojas de Cassia alata. CONCLUSIONES: La Cassia alata produce citotoxicidad en las células cancerosas A549, mediada por la activación de la caspasa 8. Este efecto puede ser atribuido al kaempferol.