Project description:Carotenoids have been known to reduce the risk of several diseases including cancer and cardiovascular. However, carotenoids are unstable and susceptible to degradation. Rhinacanthus nasutus (L.) Kurz (R. nasutus), a Chinese medicinal herb rich in carotenoids, was reported to possess vital biological activities such as anti-cancer. This study intends to isolate carotenoids from R. nasutus by column chromatography, identify and quantify by HPLC-MS, and prepare carotenoid microemulsions for determination of absolute bioavailability in rats. Initially, carotenoid fraction was isolated using 250 mL ethyl acetate poured into an open-column packed with magnesium oxide-diatomaceous earth (1:3, w/w). Fourteen carotenoids including internal standard β-apo-8'-carotenal were resolved within 62 min by a YMC C30 column and gradient mobile phase of methanol-acetonitrile-water (82:14:4, v/v/v) and methylene chloride. Highly stable carotenoid microemulsions were prepared using a mixture of Capryol(TM)90, Transcutol®HP, Tween 80 and deionized water, with the mean particle being 10.4 nm for oral administration and 10.7 nm for intravenous injection. Pharmacokinetic study revealed that the absolute bioavailability of carotenoids in microemulsions and dispersion was 0.45% and 0.11%, respectively, while a much higher value of 6.25% and 1.57% were shown for lutein, demonstrating 4-fold enhancement in bioavailability upon incorporation of R. nasutus carotenoids into a microemulsion system.
Project description:Neurodegenerative diseases present an increasing problem as the world's population ages; thus, the discovery of new drugs that prevent diseases such as Alzheimer's, and Parkinson's diseases are vital. In this study, Rhinacanthin-C and -D were isolated from Rhinacanthus nasustus, using ethyl acetate, followed by chromatography to isolate Rhinacanthin-C and -D. Both compounds were confirmed using NMR and ultra-performance-LCMS. Using glutamate toxicity in HT-22 cells, we measured cell viability and apoptosis, ROS build-up, and investigated signaling pathways. We show that Rhinacanthin-C and 2-hydroxy-1,4-naphthoquinone have neuroprotective effects against glutamate-induced apoptosis in HT-22 cells. Furthermore, we see that Rhinacanthin-C resulted in autophagy inhibition and increased ER stress. In contrast, low concentrations of Rhinacanthin-C and 2-hydroxy-1,4-naphthoquinone prevented ER stress and CHOP expression. All concentrations of Rhinacanthin-C prevented ROS production and ERK1/2 phosphorylation. We conclude that, while autophagy is present in HT-22 cells subjected to glutamate toxicity, its inhibition is not necessary for cryoprotection.
Project description:The potential benefits of natural plant extracts have received attention in recent years, encouraging the development of natural products that effectively treat various diseases. This is the first report on establishing callus and cell suspension cultures of Rhinacanthus nasutus (L.) Kurz. A yellow friable callus was successfully induced from in vitro leaf explants on Murashige and Skoog medium supplemented with 1 mg/L 2,4-dichlorophenoxyacetic acid and 1 mg/L 1-naphthalene acetic acid. A selected friable callus line was used to establish the cell suspension culture with the same medium. The antioxidant assays showed that the leaf- and ethanolic-suspension-cultured cell (SCC) extracts exhibited high antioxidant potential. In addition, the in vitro cytotoxicity revealed by the MTT assay demonstrated potent antiproliferative effects against the oral cancer cell lines ORL-48 and ORL-136 in a dose-dependent manner. Several groups of compounds, including terpenoids, phenolics, flavonoids, quinones, and stilbenes, were identified by UHPLC-QToF-MS, with the same compounds detected in leaf and SCC extracts, including austroinulin, lucidenic acid, esculetin, embelin, and quercetin 3-(2″-p-hydroxybenzoyl-4″-p-coumarylrhamnoside). The present study suggests the value of further investigations for phytochemical production using R. nasutus cell suspension culture.
Project description:BackgroundOxidative stress and nonenzymatic protein glycation lead to serious diabetic complications that increase the risk of mortality. Rhinacanthus nasutus leaf crude extracts are previously reported for their antidiabetic, antiglycation, and antioxidant potential.ObjectiveThe present study was performed to prepare a standardized rhinacanthins-rich extract (RRE) and evaluate its superoxide scavenging and antiglycation effects as compared to its marker compounds, namely, rhinacanthin-C (RC), rhinacanthin-D (RD), and rhinacanthin-N (RN).Materials and methodsRRE was obtained by microwave-assisted green extraction along with a simple step of fractionation using Amberlite® column. RC, RD, and RN were isolated from the RRE using silica gel column chromatography. Superoxide scavenging activity was performed by cyclic voltammetry, and fructose-mediated human serum albumin glycation model was used for antiglycation activity. In silico studies were conducted to identify the structure-activity relationships of rhinacanthins.ResultsOn the basis of kinetic measurements, RRE exhibited the most potent antioxidant activity via ErCi mechanism, with a 50% inhibitory concentration (IC50) value of 8.0 μg/mL, antioxidant capacity of 39439 M-1, and binding constant of 45709 M-1. Antiglycation assay showed that RRE exhibited almost equivalent glycation inhibitory effect to that of RC, with IC50 values of 39.7 and 37.3 μg/mL, respectively, but higher than that of RD (IC50 of 50.4 μg/mL), RN (IC50 of 89.5 μg/mL), as well as the positive control, rutin (IC50 of 41.5 μg/mL).ConclusionsThe potent superoxide scavenging and albumin glycation inhibitory effect of RRE rationalized its therapeutic application in various chronic diseases, especially in the complications of diabetes.SummaryRhinacanthins-rich extract (RRE) exhibited potent superoxide scavenging activityRRE and rhinacanthin-C showed remarkable and comparable antiglycation effectRhinacanthins exhibited antiglycation activity by masking specific residues of albumin. Abbreviations used: RRE: Rhinacanthins-rich extract; RC: Rhinacanthin-C; RD: Rhinacanthin-D; RN: Rhinacanthin-N; IC50: 50% inhibitory concentration; Kao: Antioxidant activity coefficient; Kb: Binding constant; ErCi: Reversible electron transfer followed by an irreversible chemical reaction; DM: Diabetes mellitus; AGEPs: Advanced glycation end products; NMR: Nuclear magnetic resonance; HPLC: High-performance liquid chromatography; CV: Cyclic voltammetry; DMSO: Dimethyl sulfoxide; Ipa: Anodic peak current; Ipc: Cathodic peak current; HSA: Human serum albumin; MOE: Molecular operating environment; PASSonline: Online prediction of activity spectra for substances.
Project description:Nanotechnology is gaining momentum due to its ability to transform metals into nanoparticles. The synthesis, characterization, and applications of biologically synthesized nanomaterials have become an important branch of nanotechnology. Plant extracts are a cost-effective, ecologically friendly, and efficient alternative for the large-scale synthesis of nanoparticles. In this study, silver nanoparticles (AgNps) were synthesized using Rhinacanthus nasutus leaf extract. After exposing the silver ions to the leaf extract, the rapid reduction of silver ions led to the formation of AgNps in solution. The synthesis was confirmed by ultraviolet-visible spectroscopy, Fourier transform infrared spectroscopy, and transmission electron microscopy. The in vitro antimicrobial activity of the AgNps synthesized using R. nasutus leaf extract was investigated against Bacillus subtilis, Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumonia, Escherichia coli, Aspergillus niger, and Aspergillus flavus using a disc diffusion method. The AgNps showed potential activity against all of the bacterial strains and fungal colonies, indicating that R. nasutus has the potential to be used in the development of value-added products in the biomedical and nanotechnology-based industries.
Project description:Garcinia spp. belongs to the family Clusiaceae has been traditionally used for the treatment of many ailments including the liver damage. Garcinia dulcis found in North Eastern region of Assam; India can be a potential candidature to combat different ailments. Objective: The present work has been designed in such a way to appraisal the antioxidant and hepatoprotective activity of fruit rind extract of this plant.The antioxidant activity was investigated through the various in vitro models, namely, 2,2-diphenyl-1-picrylhydrazine, 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid, nitrite oxide. Phytochemical investigation for total phenolic and flavonoids contents were carried out by standard protocol. For the evaluation of hepatoprotective activity, albino Wistar rats were divided into five groups, five animals per group and activity was determined by measuring the contents of liver function marker enzymes such as serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), serum alkaline phosphatase (ALP), and biochemical parameter, that is, Bilirubin and total protein. Histopathology observation of liver sections was conducted.Phytochemical investigation revealed the presence of both phenolic and flavonoid groups in the extract in a significant amount. Antioxidant activity of the plant extract was observed in all models and percentage of inhibition was dose-dependent. Intoxicated with carbon tetrachloride, elevated the liver function enzymes, bilirubin, and suppressed the production of total protein. Pretreatment with the extract decreased the SGOT, SGPT, ALP, and bilirubin level significantly and increased the production level of total protein in a dose-dependent manner. The histopathological observation supported the hepatoprotective potentiality of the extract.The results indicate that fruit rind part of G. dulcis is nontoxic and the plant can utilize as an antioxidant source. The plant has a protective agent for liver damages and other diseases caused by free radicals.In vitro antioxidant and in vivo hepatoprotective activity was evaluatedMethanolic extract was subjected to quantify the both phenolic and flavonoid contents. The extract showed the significant amount of both phenolic and flavonoids contents. The extract showed the free radical scavenging activity in 2,2-diphenyl-1-picrylhydrazine, 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid and nitrite oxide modelsThe extract was administrated against the carbon tetrachloride intoxicated animal models to evaluate the hepatoprotective activity by determining the level of liver marker enzymes such as serum glutamate pyruvate transaminase, SGOT, alkaline phosphatase and biochemical parameter such as protein and bilirubin. Pretreatment with the extract reversed the elevated level of the enzymes and increased the protein level in a dose-dependent mannerThe histopathological observations of the liver sections supported the hepatoprotective activity of the extractThe present study revealed that the Garcinia dulcis extract is a good candidature for preventing liver damage and other disease caused by free radicals. Abbreviations Used: DPPH: 2,2-diphenyl-1-picrylhydrazine, ABTS: 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid, SGOT: Serum glutamate oxaloacetate transaminase, SGPT: Serum glutamate pyruvate transaminase, ALP: Serum alkaline phosphatase.
Project description:Oroxylum indicum (L.) Kurz, a medicinal plant, shows numerous pharmacological properties which may be attributed to the bioactive compounds produced by O. indicum or due to associated endophytes. In the present study, leaf of O. indicum was evaluated for the presence of associated fungal endophytes, and antioxidant and cytotoxic activities of bioactive compounds produced from them. Using culture-dependent approach, eight fungal endophytes belonging to five different genera were identified. Two endophytes Daldinia eschscholtzii and Ectophoma multirostrata have been reported for the first time from the leaf of O. indicum plant. High-performance thin-layer chromatography (HPTLC) of ethyl acetate (EA) extract of isolated fungal endophytes showed a distinct fingerprinting profile in EA extract of Colletotrichum gloeosporioides. Among identified endophytes, EA extract of C. gloeosporioides showed significant antioxidant activity against DPPH free radical, superoxide anion radical, nitric oxide radical and hydroxyl radical with EC50 values of 22.24±1.302 μg/mL, 67.46±0.576 μg/mL, 80.10±0.706 μg/mL and 61.55±1.360 μg/mL, respectively. EA extract of C. gloeosporioides exhibited potential cytotoxicity against HCT116, HeLa and HepG2 cancer cell lines with IC50 values of 76.59 μg/mL, 176.20 μg/mL and 1750.70 μg/mL, respectively. A comparative HPTLC fingerprinting and the antioxidant activity of C. gloeosporioides associated with two different hosts (leaf of O. indicum and dead twigs of other plant) showed that C. gloeosporioides produces bioactive compounds in a host-dependent manner.
Project description:Nigella sativa L. (NS) is an herbaceous plant, possessing phytochemicals of therapeutic importance. Thymoquinone is one of the active phytochemicals of NS that confers noteworthy antioxidant properties. Sodium azide, an agent of abiotic stress, can modulates antioxidant system in plants. In the present investigation, sodium azide (0, 5 µM, 10 µM, 20 µM, 50 µM, 100 µM and 200 µM) doses administered to the in vitro NS callus cultures for production/modification of secondary metabolites with augmented activity. 200 µM sodium azide treated NS callus exhibited maximum peroxidase activity (1.286 ± 0.101 nanokatal mg-1 protein) and polyphenol oxidase activity (1.590 ± 0.110 nanokatal mg-1 protein), while 100 µM sodium azide treated NS callus for optimum catalase activity (1.250 ± 0.105 nanokatal mg-1 protein). Further, 200 µM sodium azide treated NS callus obtained significantly the highest phenolics (3.666 ± 0.475 mg g-1 callus fresh weight), 20 µM sodium azide treated NS callus, the highest flavonoids (1.308 ± 0.082 mg g-1 callus fresh weight) and 100 µM sodium azide treated NS callus, the highest carotenes (1.273 ± 0.066 mg g-1 callus fresh weight). However, NS callus exhibited a decrease in thymoquinone yield/content vis-à-vis possible emergence of its analog with 5.3 min retention time and an increase in antioxidant property. Treatment with 200 µM sodium azide registered significantly the lowest percent yield of callus extract (4.6 ± 0.36 mg g-1 callus fresh weight) and thymoquinone yield (16.65 ± 2.52 µg g-1 callus fresh weight) and content (0.36 ± 0.07 mg g-1 callus dry weight) and the highest antioxidant activity (3.873 ± 0.402%), signifying a negative correlation of the former with the latter. DNA damage inhibition (24.3 ± 1.7%) was recorded significantly maximum at 200 µM sodium azide treatment. Sodium azide treated callus also recorded emergence of a new peak at 5.3 min retention time (possibly an analog of thymoquinone with augmented antioxidant activity) whose area exhibits significantly negative correlation with callus extract yield and thymoquinone yield/content and positive correlation with antioxidant activity and in vitro DNA damage inhibition. Thus, sodium azide treatment to NS callus confers possible production of secondary metabolites or thymoquinone analog (s) responsible for elevated antioxidant property and inhibition to DNA damage. The formation of potent antioxidants through sodium azide treatment to NS could be worthy for nutraceutical and pharmaceutical industries.
Project description:Ferulic acid is a derivative of cinnamic acid showing efficacious anti-oxidant activity. It catalyzes the stable phenoxy radical formation, upon absorption of ultraviolet light, giving the strength to ferulic acid for terminating free radical chain reactions. Ultraviolet rays are one of the most dangerous factors that daily assault the skin, causing excessive generation of reactive oxygen species (ROS), which are regarded to be important contributors to a variety of cutaneous alterations. The skin possesses endogenous antioxidant defense systems, but the excess of ROS leads to an oxidant-antioxidant imbalance. Although ferulic acid is daily introduced in human organism with the diet, its bioavailability after oral administration is poor, particularly in the skin. The aim of this investigation was to evaluate three types of emulsions (W/O/W multiple emulsions and two simple emulsions) as suitable formulations for topical application of the active compound. In vitro studies were performed to investigate the stability and release profiles of these systems. Multiple emulsions showed great stability and the best ability to carry and release ferulic acid. In vivo evaluations highlighted their best capability to treat UV-B-induced erythema. These findings suggested multiple emulsions as an innovative and more efficient vehicle for topical application of ferulic acid.
Project description:Olive pomace (OP) is a valuable food byproduct that contains natural phenolic compounds with health benefits related to their antioxidant activities. Few investigations have been conducted on OP from the United States while many studies on European OP have been reported. OP of Arbequina, the most common cultivar from California, was collected and extracted by water, 70% methanol and 70% ethanol, followed by purification using macroporous absorbing resin. Results showed that the extractable total phenolic content (TPC) was 36-43 mg gallic acid equivalents (GAE)/g in pitted, drum-dried defatted olive pomace (DOP), with major contributions from hydroxytyrosol, oleuropein, rutin, verbascoside, 4-hydroxyphenyl acetic acid, hydroxytyrosol-glucoside and tyrosol-glucoside. Macroporous resin purification increased TPC by 4.6 times the ethanol crude extracts of DOP, while removing 37.33% total sugar. The antioxidant activities increased 3.7 times Trolox equivalents (TrE) by DPPH and 4.7 times TrE by ferric reducing antioxidant power (FRAP) in the resin purified extracts compared to the ethanol crude extracts. This study provided a new understanding of the extraction of the bioactive compounds from OP which could lead to practical applications as natural antioxidants, preservatives and antimicrobials in clean-label foods in the US.