Assessment of Anti-Inflammatory and Antioxidant Activities of a Proprietary Preparation of Quercetin-Rutin Blend (SophorOx™) in Exercised Rats.

Objectives: Flavonoids comprise a huge class of phenolic compounds widely distributed throughout the plant kingdom. Although quercetin and rutin have been studied individually for their therapeutic value, the synergistic effect of combining the two has previously not been measured. The objective of this trial was to evaluate the anti-inflammatory and antioxidant properties of both quercetin and rutin when combined in the form of SophorOx™ (a proprietary preparation of quercetin-rutin) in exercised rats. Methods: Sprague-Dawley rats were orally administered SophorOx™ at 500 mg·kg-1·b.w. and subjected to daily exercise on a fabricated treadmill for 4 weeks. A total of 24 animals were randomly divided into four groups. All the animals were examined for body weight, feed consumption, signs of clinical abnormalities, and morbidity. In addition, serum collected on days 8, 15, 22, and 29 were measured for the liver function test (LFT), random blood sugar (RBS), inflammatory markers C-reactive protein (CRP), oxidative stress markers (8-isoprostane (8-iso-PGF2α), malondialdehyde (MDA), 8-hydroxydeoxyguanosine (8-OHdG), and cytokine levels interleukin-1ß (IL-1ß), interleukin 6 (IL-6), and tumor necrosis factor-α (TNF-α)) by the ELISA method. Results: Rats that received SophorOx™ showed no signs of adverse effects, and no significant changes were observed in body weight, feed consumption, liver enzymes, and blood glucose levels. The exercise-treated rats administered with SophorOx™ exhibited a significant reduction in oxidative and inflammatory marker levels, viz., CRP (113.32 ng·mL-1) and oxidative stress markers 8-OHdG (19.32 pg·mL-1), MDA (1.06 nmol·mL-1), 8-iso-PGF2α (1.29 ng·mL-1), IL-1ß (0.77 pg·mL-1), and IL-6 (317.14 pg·mL-1) in comparison to those rodents that were exercised without SophorOx™. Conclusion: Oral administration of SophorOx™ significantly reduced oxidative stress and inflammatory marker levels when measured in the rodents subjected to high-intensity exercise.

Effect of Broad-Spectrum Hemp Extract on Neurobehavioral Activity on the Immobilization Stress-Induced Model in Sprague Dawley Rats.

Background: Broad-spectrum hemp extract is expected to be a promising new intervention for managing stress and anxiety. Research has shown that the cannabinoids found in Cannabis sativa, such as cannabidiol (CBD), tetrahydrocannabinol (THC), and cannabigerol (CBG), possess anxiolytic properties that can positively impact mood and stress. Methods: In the current study, a broad-spectrum, nondetectable THC hemp extract with other minor cannabinoids (broad-spectrum hemp extract) was administered at 28 mg/kg·bw to evaluate its anxiolytic properties. This was performed using various behavioural models and biomarkers for oxidative stress. In addition, a 300 mg/kg·bw of Ashwagandha root extract was also incorporated to compare its effects on relieving stress and anxiety. Results: The decreased levels of lipid peroxidation were measured in broad-spectrum hemp extract (36 nmol/ml), Ashwagandha (37 nmol/ml), and induction control (49 nmol/ml) treated groups of animals. The levels of 2-AG decreased in the broad-spectrum hemp extract (1.5 ng/ml), Ashwagandha (1.2 ng/ml), and induction control (2.3 ng/ml) treated groups of animals. The levels of FAAH decreased in broad-spectrum hemp extract (1.6 ng/ml), Ashwagandha (1.7 ng/ml), and induction control (1.9 ng/ml) treated groups of animals. The levels of catalase increased in broad-spectrum hemp extract (35 ng/ml), Ashwagandha (37 ng/ml), and induction control (17 ng/ml) treated groups of animals. Similarly, increased levels of glutathione were found in broad-spectrum hemp extract (30 ng/ml), Ashwagandha (27 ng/ml), and induction control (16 ng/ml) treated groups of animals. Conclusion: Based on the results of this study, it can be concluded that broad-spectrum hemp extract inhibited the biomarkers for oxidative stress. Also, certain behavioural parameters showed improvements with respect to both the ingredient administered groups.

Safety Assessment of L-ß-Aminoisobutyric Acid (L-BAIBA): Subchronic Toxicity Study in Sprague Dawley Rats.

L-3-Aminoisobutyric acid (L-BAIBA) is an endogenous compound in human metabolism when thymine and valine undergo catabolism. L-BAIBA represents one of the two isomers of BAIBA in biological systems. BAIBA has been shown to reduce body fat percentage via an increase in fatty acid oxidation and a decrease in hepatic lipogenesis. However, no toxicological effects of L-BAIBA in animals or humans have been established. The present study was designed to evaluate the safety and toxic potentials of this compound, where L-BAIBA was administered orally to Sprague Dawley rats at 100, 300, and 900 mg/kg/day for 90 days. No treatment-related adverse effects were observed in any of the treatment groups. Based on the results, the No-Observed-Adverse-Effect Level (NOAEL) of L-BAIBA was 900 mg/kg/day.

Toxicological evaluation of a fish oil concentrate containing Very Long Chain Fatty Acids.

Very long chain fatty acids (VLCFA) have a chain length ≥24 carbons. Fish contain low levels of these fatty acids. A commercial oil called EPAX® Evolve 05 with an up-concentration of VLCFAs of approximately 10 times, has been developed as a dietary supplement by Epax Norway AS. A series of toxicological studies were performed using mice and rats to determine the safety and toxicity of repeat dosing with a gavage administered VLCFA formulation. The results suggest transient lipid accumulation in kidneys and liver. Lipid accumulation was seen with the test item and with the soya control but was not dose related. Liver and kidney lipid accumulation, whilst present in 14- day repeat dose study, was absent in a 90-day rat study. No treatment-effect was seen in urine analysis in any of the studies. No treatment-related effects were seen with a functional observation battery, ophthalmological examination, haematology, urine analysis, oestrus cycle, thyroid hormones, organ weight, or histopathology. In the 90-day study the liver enzymes ALP, AST and ALT were statistically significantly increased with test item but within control values. There were no associated histological findings in the liver suggesting there was no toxic effect and the normalisation of values for all liver enzymes in the recovery groups suggests an adaptive response rather than a prevailing toxic response. The no-observed-adverse-effect level (NOAEL) was determined as 1200 mg VLCFA/kg b.w./day.

Subchronic toxicological evaluation of EnXtra™ (standardised extract of Alpinia galanga rhizome) in rats.

OBJECTIVES: Alpinia galanga is a commonly used ingredient in Asian food and traditional medicine. But an extract of the rhizome had never been used commercially in food supplements and functional foods. There is some evidence of safety and tolerability in humans for a proprietary A. galanga rhizome extract (EnXtra™) and it is Generally Recognised as Safe (GRAS) in the US already. However, this botanical ingredient has not been evaluated for its subchronic toxicity in rats to confirm its safety in wider food applications. METHODS: Sprague Dawley rats were orally administered the test item for 90 days by following OECD (Test Guideline: 408), with a recovery period of 28 days. Cumulative effects and No Observed Adverse Effect Level (NOAEL) were estimated. EnXtra™ was administered orally at 0, 1,000, 2,000 and 3,000 mg kg-1 body weight (b. wt.) with additional vehicle and high dose recovery groups. Observations included clinical signs, haematology, clinical chemistry, gross pathology and histopathology. RESULTS: On terminal sacrifice, no treatment-related adverse effects were observed viz., clinical signs, mortality, body weight changes and feed consumption parameters. Haematology, clinical biochemistry and thyroid hormone levels were within the normal range. Further, no treatment-related gross and microscopic pathological lesions were observed across the treatment groups. CONCLUSIONS: Based on the results of the toxicological evaluation, NOAEL of A. galanga rhizome extract (AGRE) was fixed at 3,000 mg kg-1 b. wt. per day and ADI of 1800 mg day-1 in the case of humans.

Assessment of anti-inflammatory and antioxidant activity of quercetin-rutin blend (SophorOx™) - an invitro cell based assay.

OBJECTIVES: Quercetin & Rutin, are bioactive compounds that are widely used for various therapeutic properties. There's been growing interest in the biological activities of these polyphenols belonging to the class of flavonoids known to have various health benefits. Quercetin is now popularly recognized as a phytochemical remedy for a plethora of disease groups such as metabolic syndrome (more specifically diabetes), obesity/weight management and mood disorders. Due to its unique chemical structure, the most prominent property of Quercetin is probably its antioxidant capability. It acts as a free radical scavenger to form resonance-stabilized phenoxyl radicals. Certain in vitro studies have also shown quercetin to have anti-viral, anti-carcinogenic and platelet aggregation properties. Rutin has also been shown to exert diverse biological effects such as anti-tumor activities, reduction of inflammatory cytokines and antimicrobial activities. The current study was designed to further confirm the antioxidant and anti-inflammatory property of a Quercetin-Rutin blend (SophorOx™). METHODS: The analysis was performed in a cell-based assay using RAW 264.7 macrophage cell line. SophorOx™ was screened for cytotoxicity using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) to obtain optimum concentrations for experimental assays. SophorOx™ was measured for pro-inflammatory cytokine levels (TNF-α & IL-6) and nitric oxide (NO) levels. Additionally, ROS (reactive oxygen species) levels in RAW cells were estimated using a cell-permeant reagent 2'7'-dichlorofluorescein diacetate (DCFH-DA). RESULTS: SophorOx™ at 10 µM concentration, exhibited an anti-inflammatory property with significant inhibitory levels of TNF-α (∼28.25%) and IL-6 (∼32.25%). SophorOx™ at similar concentrations reduced nitric oxide levels to 70.55% in lipopolysaccharide (LPS) stimulated RAW 264.7 cells. Raw 264.7 cells stimulated with LPS exhibited a significant increase in intracellular ROS and this was significantly reduced (78.2% reduction) at lower concentrations (0.3 µM) of SophorOx™. CONCLUSIONS: The anti-inflammatory effects of SophorOx™ were investigated in LPS stimulated RAW 264.7 macrophages. Data suggests, that SophorOx™ reduced levels of nitric oxide, intracellular ROS and pro-inflammatory cytokines (TNF-α & IL-6) at low concentrations without affecting the viability of RAW cells. Present invitro trial suggests that SophorOx™ is a potent antioxidant and anti-inflammatory agent and displays a prominent ability to block mediators of oxidative stress and inflammation.

Development of an antidiabetic formulation (ADJ6) and its inhibitory activity against α-amylase and α-glucosidase.

There has recently been much advancement in the diagnosis, treatment, and research of metabolic disorders, especially diabetes. Current research around the world is focused on finding an alternative source of treatment from natural resources for diabetic management, apart from the available synthetic medicines. The present study is a preliminary study of a polyherbal formulation using edible natural resources and an assessment of its antidiabetic activity. The formulation was screened for its phytochemical constituents, total phenols, flavonoids, and vitamin C content. It was also analyzed for its inhibitory effect against the digestive enzymes α-amylase and α-glucosidase, compared with the standard drug acarbose. The formulation showed the presence of major constituents such as steroids, cardiac glycosides, phenols, flavonoids, and saponins. It also had a high level of phenols (340 ± 2.5 mg/g), flavonoids (235.4 ± 8.3 mg/g), and vitamin C (470.8 ± 16.6 mg/g), and showed a half-maximal inhibitory concentration (IC50) value of 0.41 ± 0.03 mg/mL and 0.51 ± 0.01 mg/mL for amylase and glucosidase, respectively. The results showed that ADJ6 had a significant inhibitory activity on α-amylase and α-glucosidase; however, its inhibitory activity was less than that of acarbose. The plants that are formulated in ADJ6 possess potent antidiabetic activity. Thus, we found that ADJ6 is a potent lead for effective diabetic management; however, an evaluation of the formulation must be illustrated using an in vivo model.

Development of an antidiabetic polyherbal formulation (ADPHF6) and assessment of its antioxidant activity against ROS-induced damage in pUC19 and human lymphocytes - an in vitro study.

BACKGROUND: Polyherbalism, an alternative natural-based therapy for various disorders, has been quoted about 1,300 years before in Sharangdhar Samhita. Herbal-based combination therapy stages a vital role for the treatment of type 2 diabetes mellitus (T2DM) and associated complications. The present study aims at developing an Ayurvedic-based polyherbal formulation (ADPHF6) and the assessing its antidiabetic and antioxidant property. METHODS: ADPHF6 polyherbal formulation was measured for phytochemical components by qualitative methods. The polyherbal formulation was quantitatively estimated for its phytochemical constituents, i. e. total phenol and flavonoid content. Further, the antioxidant property of ADPHF6 formulation was evaluated by 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) radical scavenging assay, hydrogen peroxide radical scavenging assay and metal chelating assay. α-Amylase and α-glucosidase inhibitory activities of polyherbal formulation were also assessed. ADPHF6 was further analysed for its protective antioxidant property against reactive oxygen species (ROS‾)-induced damage in human lymphocyte DNA and pUC19 plasmid. RESULTS: ADPHF6 polyherbal formulation revealed the presence of phytochemical constituents such as alkaloids, flavonoids, phenols, tannins, terpenoids, saponins and cardiac glycosides in significant levels. Further, it also measured the higher levels of total phenols (473.3±3.05 mg/g) and flavonoid (664±5.29 mg/g) content. Polyherbal formulation also exhibited IC50 values of 49.9±0.15, 65.1±0.10 and 60.1±0.05 mg/mL for 2,2- diphenyl-1-picryl-hydrazyl-hydrate (DPPH), hydrogen peroxide (H2O2) and Fe2+ radical scavenging activities, respectively. ADPHF6 revealed an inhibitory activity (IC50) of 0.67±0.01 and 0.81±0.01 mg/mL for α-amylase and glucosidase, respectively. Pre-treated human peripheral blood lymphocytes with ADPHF6 aqueous extract illustrated enhanced protection against ROS-mediated damage as compared with post-treated groups. DNA nicking assay rendered protective activity against the OH¯ radical-induced DNA damage in supercoiled pUC19 plasmid. CONCLUSIONS: Our present study demonstrates that ADPHF6 offers potent inhibitory activity against free radicals as well as digestive enzymes. However, studies should be conducted using in vivo model to further elucidate the effect against free radicals and its anti-hyperglycaemic activity in the management of non-insulin-dependent diabetes.