Phenolic rich Cocos nucifera inflorescence extract ameliorates inflammatory responses in LPS-stimulated RAW264.7 macrophages and toxin-induced murine models.

Anti-inflammatory and antinociceptive effects of the acetone extract of Cocos nucifera (CnAE), an important ingredient in several traditional drugs, have been studied using different in vitro and in vivo models. CnAE did not show any observable toxicity in RAW264.7 macrophages by MTT assay. The calorimetric analysis (total COX, 5-LOX, MPO, iNOS and NO), ELISA (IL-1ß, IL-6, TNF-α and PGE2) and qRT-PCR (IL-1ß, IL-6, TNF-α and NF-κB) were performed in LPS-induced RAW264.7 macrophages. Phosphorylation of NF-κBp65 and IκB was determined by western blotting. CnAE (100 µg/mL) remarkably inhibited total COX (68.67%) and 5-LOX (63.67%) activities, and subsequent release of iNOS, NO and PGE2 (p ≤ 0.05) in RAW264.7 cells treated with LPS. ELISA showed CnAE markedly decreased the level of pro-inflammatory cytokines IL-1ß (p ≤ 0.001), IL-6 (p ≤ 0.001) and TNF-α (p ≤ 0.001) in LPS treated RAW264.7 cells. CnAE (100 µg/mL) also significantly down-regulated the mRNA expressions of pro-inflammatory cytokines (IL-1ß, p ≤ 0.05; IL-6, p ≤ 0.01 and TNF-α, p ≤ 0.001) and NF-κB (p ≤ 0.001) against LPS-induction. Moreover, LPS-induced phosphorylation of IκB-α and NF-κB p65 was significantly inhibited by CnAE (100 µg/mL). In vivo anti-inflammatory studies showed that CnAE (400 mg/kg) significantly inhibited carrageenan-induced acute paw oedema (59.81%, p ≤ 0.001) and formalin-induced chronic paw oedema (52.90%, p ≤ 0.001) in mice. CnAE at a dose of 400 mg/kg also showed a significant anti-nociceptive effect on acetic acid-induced writhing (48.21%, p ≤ 0.001) and Eddy's hot plate methods. These findings suggest that CnAE has significant anti-inflammatory and anti-nociceptive properties, mainly attributed to the inhibition of NF-κB/IκB signalling cascade.

High-Performance Liquid Chromatography (HPLC) as a Tool for Standardization of Complex Herbal Drugs.

Background: Herbal formulations have reached tremendous acceptability as therapeutic agents for several diseases mainly due to the indiscriminate use of modern medicine such as antibiotics, steroids and other synthetic drugs. The increasing popularity in plant-based drugs is leading to a fast growing market for plant-based drugs pharmaceuticals, nutraceuticals, functional foods, and even cosmeceuticals. Objective: The development of authentic analytical methods for complex herbal drugs, especially poly herbal formulations which can reliably profile the phytochemical composition, including quantitative analyses of marker/bioactive compounds and other major constituents, is a major challenge to scientists. Standardization is an important step for the establishment of a consistent biological activity, a consistent chemical profile, or simply a quality assurance program for production and manufacturing of herbal drugs. Methods/Results: HPLC as a tool has been widely used in the standardization of complex herbal drugs because of its ability to estimate the presence of active (chemical or biological) markers both qualitatively and quantitatively. Conclusions: An overview of various HPLC techniques that can be used for standardization of herbal drugs has been presented.

Novel mechanism of modulating natural antioxidants in functional foods: involvement of plant growth promoting Rhizobacteria NRRL B-30488.

The significance of plant growth-promoting rhizobacteria (PGPR) mediated increase in antioxidant potential in vegetables is yet unknown. The plant growth-promoting bacterium Bacillus lentimorbus NRRL B-30488 (B-30488) mediated induction of dietary antioxidant in vegetables ( Trigonella foenum-graecum, Lactuca sativa, Spinacia oleracea, and Daucus carota) and fruit ( Citrus sinensis) after minimal processing (fresh, boiled, and frozen) was tested by estimating the total phenol content, level of antioxidant enzymes, and 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide scavenging activities along with integral radical scavenging capacity by photochemiluminescence assay and inhibition of lipid peroxidation. Minimal processing of vegetables showed that T. foenum-graecum had the highest phenol content in B-30488-treated plants followed by L. sativa, D. carota, and S. oleracea. Thermally treated vegetables T. foenum-graecum (26-114.5 GAE microg mg (-1)) had an exceptionally high total phenolic content, followed by D. carota (25.27-101.32 GAE microg mg (-1)), L. sativa (23.22-101.10 GAE microg mg (-1)), and S. oleracea (21.87-87.57 GAE microg mg (-1)). Among the vegetables and fruit used in this study for enzymatic estimation, induction of antioxidant enzymes, namely, polyphenol oxidase (PPO), ascorbate peroxidase (APX), catalase (CAT), and superoxidase dismutase (SOD), was observed in edible parts of T. foenum-graecum, L. sativa, S. oleracea, and D. carota, after inoculation with B-30488. The scavenging capacity of the vegetables treated with B-30488 against DPPH and superoxide anion radical activity was found to be significantly high as compared to nontreated control. Mild food processing had no adverse effect on radical scavenging capacity. Photochemiluminescence also ascertains the above findings. The ability of the plant extracts to protect against lipid peroxidation and its ability to prevent oxidation of reduced glutathione (GSH) was measured in rat liver homogenate, and the results suggested that the inoculated plant exhibited better activity in all of the screened plants. Significant increases in shoot length, root length, and dry weight, averaging 164, 132, and 135% in T. foenum-graecum, 174, 141, and 156% in L. sativa, 129, 141, and 59%, in S. oleracea, and 125, 146, and 42% in D. carota, respectively, over untreated controls, were attained in greenhouse trials. To the best of the authors' knowledge, this is the first report of PGPR-mediated induction of antioxidant enzyme activity (PPO, APX, CAT, and SOD) along with the antioxidant activity of the extracts in both in vitro (DPPH radical scavenging and superoxide scavenging) and ex vivo conditions using the rat liver tissue (percent inhibition of lipid peroxidation and prevention of oxidation of GSH) and phenolic content. The results demonstrate the PGPR-mediated induction of antioxidant level in vegetables and fruit controls oxidative damage even after minimal processing and thus is indicative of its potential as a viable substitute of synthetic antioxidants.

Hepatoprotective effects of rubiadin, a major constituent of Rubia cordifolia Linn.

The hepatoprotective effects of rubiadin, a major constituent isolated from Rubia cordifolia Linn., were evaluated against carbon tetrachloride (CCl4)-induced hepatic damage in rats. Rubiadin at a dose of 50, 100 and 200 mg/kg was administered orally once daily for 14 days. The substantially elevated serum enzymatic activities of serum glutamic oxaloacetic transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), serum alkaline phosphatase (SALP) and gamma-glutamyltransferase (gamma-GT) due to carbon tetrachloride treatment were dose dependently restored towards normalization. Meanwhile, the decreased activities of glutathione S-transferase and glutathione reductase were also restored towards normalization. In addition, rubiadin also significantly prevented the elevation of hepatic malondialdehyde formation and depletion of reduced glutathione content in the liver of CCl4 intoxicated rats in a dose dependent manner. Silymarin used as standard reference also exhibited significant hepatoprotective activity on post treatment against carbon tetrachloride induced hepatotoxicity in rats. The biochemical observations were supplemented with histopathological examination of rat liver sections. The results of this study strongly indicate that rubiadin has a potent hepatoprotective action against carbon tetrachloride induced hepatic damage in rats.

Antioxidant activity of Desmodium gangeticum and its phenolics in arthritic rats.

Total alcoholic extract of Desmodium gangeticum, which exhibited significant anti-inflammatory activity, was evaluated for the possible mode of action by studying its antioxidant potential in adjuvant-induced arthritic rats. Activity guided fractionation and isolation were carried out. The phenolics fraction showed maximum potency. Solid phase extraction followed by preparative HPLC of the active phenolic fraction yielded for the first time two potent antioxidant compounds, caffeic acid and chlorogenic acid, from this plant. The biological antioxidant defense system, involving superoxide dismutase, glutathione and catalase, showed a significant increase with their levels close to the normal control with a decrease in the lipid peroxide content upon administration of D. gangeticum extract (100 mg kg(-1)) and its phenolics (50 mg kg(-1)) in arthritic rats, thereby indicating the extracts antioxidant property under arthritic conditions.

Free radical scavenging potential of Saussarea costus.

Saussurea costus (Falc.) Lipschitz from the family Asteraceae is an important medicinal drug, the roots of which are widely used in folk medicine. The antioxidant activity of the plant has been studied using its ability to scavenge DPPH, nitric oxide, superoxide radicals along with its ability to inhibit lipid peroxidation and GSH oxidation. The 1 mg mL(-1) extract had antioxidant activity with 85.2% reduction of DPPH and a 72.7% decrease in lipid peroxidation. It showed maximum inhibition of superoxide radical of 66.0%, and 58.4% inhibition of nitric oxide formation. The concentration of chlorogenic acid was 0.027% in the extract of S. costus. Thus, the therapeutic activity of the plant may be due to its antioxidant activity, probably as a result of the presence of chlorogenic acid.

Antioxidant potential of Anogeissus latifolia.

Anogeissus latifolia is widely used in the Indian indigenous system of medicine and is reported to contain leucocyanidins and tannoid principles like ellagic acid and its derivatives. In view of its wide use and its chemical composition, this study was aimed at examining the antioxidant activity of the extract of A. latifolia. The extract was studied for total antioxidant capacity, hydrogen-donating ability, nitric oxide, superoxide scavenging activity, hydrogen peroxide decomposition activity along with lipid peroxidation. Integral antioxidative capacity was determined by chemiluminescence assay. The extract was also studied for lipid peroxidation assay by thiobarbituric acid-reactive substances (TBARS) method using rat liver homogenate. The results indicate that A. latifolia extract has potent antioxidant activity. Also to ascertain the possible reason for the potent activity, percentage of gallic acid was estimated and was found to be 0.95%, which could be one of the reasons for potent antioxidant activity exhibited by the plant.

Healing potential of Anogeissus latifolia for dermal wounds in rats.

Wound healing potential of ethanolic extract of Anogeissus latifolia bark (ALE) for treatment of dermal wounds in rats was studied on excision and incision wound models. HPTLC of the total extract was recorded for the purpose of standardization. Various parameters of incision wound, viz. epithelization period, scar area, tensile strength and hydroxyproline measurements along with wound contraction, were used to evaluate the effect of A. latifolia on wound healing. The results obtained indicate that A. latifolia accelerates the wound healing process by decreasing the surface area of the wound and increasing the tensile strength. Nitrofurazone ointment was used as a positive control. Complete epithelization was observed within 15 days with ALE. Measurements of the healed area and the hydroxyproline level were in agreement. Antibacterial activity of ALE was studied against Gram-positive (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumoniae) compared to erythromycin and tetracycline. Moderate activity was observed against all organisms. The present study provides a scientific rationale for the traditional use of Anogeissus latifolia in the management of skin diseases such as sores, boils and itching.

Studies on the antioxidant activities of Desmodium gangeticum.

Desmodium gangeticum is herbal species which is widely used in the indigenous system of medicine and is reported to contain flavone and isoflavanoid glycosides. In view of its wide use and it's chemical composition, this study was aimed at examining the antioxidant activity of the extract of D. gangeticum. The extract was studied for diphenyl picryl hydrazyl (DPPH), nitric oxide, ferryl-bipyridyl and hypochlorous acid scavenging activity along with lipid peroxidation. Nitric oxide was generated using sodium nitroprusside and was studied using Griess reagent. In order to study the iron chelating capacity of the extract, the percentage ferryl-bipyridyl inhibition was studied. Hypochlorous acid scavenging activity was tested by measuring the inhibition of 5-thio-2-nitrobenzoic acid oxidation. The extract was also studied for lipid peroxidation assay by thiobarbituric acid-reactive substances (TBARS) method using rat brain homogenate. The results indicate that D. gangeticum extract has potent antioxidant activity.