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Quantitative Determination of Stilbenoids and Dihydroisocoumarins in Shorea roxburghii and Evaluation of Their Hepatoprotective Activity.

ABSTRACT: A simultaneous quantitative analytical method for 13 stilbenoids including (-)-hopeaphenol (1), (+)-isohopeaphenol (2), hemsleyanol D (3), (-)-ampelopsin H (4), vaticanols A (5), E (6), and G (7), (+)-α-viniferin (8), pauciflorol A (9), hopeafuran (10), (-)-balanocarpol (11), (-)-ampelopsin A (12), and trans-resveratrol 10-C-β-d-glucopyranoside (13), and two dihydroisocoumarins, phayomphenols A₁ (14) and A₂ (15) in the extract of Shorea roxburghii (dipterocarpaceae) was developed. According to the established protocol, distributions of these 15 polyphenols (1-15) in the bark and wood parts of S. roxburghii and a related plant Cotylelobium melanoxylon were evaluated. In addition, the principal polyphenols (1, 2, 8, 13-15) exhibited hepatoprotective effects against d-galactosamine (d-galN)/lipopolysaccharide (LPS)-induced liver injury in mice at a dose of 100 or 200 mg/kg, p.o. To characterize the mechanisms of action, the isolates were examined in in vitro studies assessing their effects on (i) d-GalN-induced cytotoxicity in primary cultured mouse hepatocytes; (ii) LPS-induced nitric oxide (NO) production in mouse peritoneal macrophages; and (iii) tumor necrosis factor-α (TNF-α)-induced cytotoxicity in L929 cells. The mechanisms of action of these polyphenols (1, 2, and 8) were suggested to be dependent on the inhibition of LPS-induced macrophage activation and reduction of sensitivity of hepatocytes to TNF-α. However, none of the isolates reduced the cytotoxicity caused by d-GalN.

SUBMITTER: Ninomiya K

PROVIDER: S-EPMC5343985 | biostudies-literature |

REPOSITORIES: biostudies-literature

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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:α and β asarones are natural constituents of some aromatic plants, especially species of the genus Acorus (Araceae). In addition to reports of beneficial properties of asarones, genotoxicity and carcinogenicity are also reported. Due to potential toxic effects of β-asarone, a limit of exposure from herbal products of ~2 μg/kg body weight/day has been set temporarily until a full benefit/risk assessment has been carried out by the European Medicines Agency. Therefore, it is important to monitor levels of β-asarone in herbal products. In this study, we developed a simple, rapid and validated GC-MS method for quantitative determination of asarones and applied it in 20 pediatric herbal products after detecting high concentrations of β-asarone in a product suspected to be implicated in hepatotoxicity in a 3 month old infant. Furthermore, targeted toxicological effects were further investigated in human hepatocytes (THLE-2 cells) by employing various in vitro assays, with the goal of elucidating possible mechanisms for the observed toxicity. Results showed that some of the products contained as much as 4-25 times greater amounts of β-asarone than the recommended levels. In 4 of 10 samples found to contain asarones, the presence of asarones could not be linked to the labeled ingredients, possibly due to poor quality control. Cell-based investigations in THLE-2 cells confirmed the cytotoxicity of β-asarone (IC50 = 40.0 ± 2.0 μg/mL) which was associated with significant lipid peroxidation and glutathione depletion. This observed cytotoxic effect is likely due to induction of oxidative stress by asarones. Overall, the results of this study ascertained the usability of this GC-MS method for the quantitative determination of asarones from herbal products, and shed light on the importance of controlling the concentration of potentially toxic asarones in herbal products to safeguard consumer safety, especially when the target consumers are young children. Further investigations of the toxicity of asarones are warranted.

Dataset Information

Quantitative Determination of Stilbenoids and Dihydroisocoumarins in Shorea roxburghii and Evaluation of Their Hepatoprotective Activity.

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