Heartwood Of Pterocarpus Marsupium Research Articles

Optimization of bioactivities of solvent extracts of bark and heartwood of Pterocarpus marsupium and exploration of neuroprotective effect linking to the major Phytoconstituents

The study aims to evaluate the neuroprotective properties of Pterocarpus marsupium Roxb. The bioactive optimized hydroalcohol-extracted fractions of bark (PMBHA) and heartwood (PMHHA) at 300 and 400 mg/kg were examined in rats with scopolamine-induced neurotoxicity to study oxidative, neurochemical, and neuroinflammation biomarkers. Phenol and flavonoid-rich radical scavengers in contributing total antioxidant capacities of PMBHA and PMHHA quenched peroxyl radicals and protected the cellular membrane and lipoproteins from ROS in DPPH, ORAC, and CAP-e tests. In anticholinesterase activities, PMBHA inhibited AChE (IC50: 54.40 ± 0.98 µg/mL) and BChE (IC50: 50.95 ± 0.98 µg/mL) mixed-competitively, while PMHHA inhibited cholinesterase activities (AChE: 42.23 ± 0.54, and BChE: 47.19 ± 0.65 µg/mL) competitively. Dosage at 300 and 400 mg/kg of PMBHA and PMHHA improved serum and brain oxidative indicators (MDA, SOD, CAT, and GSH), and restored proinflammatory cytokines (TNF-α, and IL-6) leading to enhanced brain cholinesterase and β-secretase levels in neurotoxic rats. GC-MS reported antioxidant, anti-inflammatory, and neuroprotective compounds of PMBHA catechol, thiamet-G, d-Gala-l-ido-octonic amide, falcarinol, D-mannose, ribitol, 1,2,4-cyclopentanetrione, 3-(2-pentenyl)-, hexadecanoic acid methyl ester, n-hexadecanoic acid, oleic acid, and 3,5-Dimethoxybenzyl alcohol, α-epi-7-epi-5-Eudesmol, 1 R,4aR,7 R,8aR)-7-(2-Hydroxypropan-2-yl)-1,4a-dimethyldecahydro naphthalen-1-ol, arctiol, 1,1,4,7-tetramethyldecahydro-1 H-cyclopropa[e]azulene-4,7-diol, 2-Propen-1-ol, 3-(2,6,6-trimethyl-1-cyclohexen-1-yl), 4-t-Butyl-2-[4-nitrophenyl]phenol in PMHHA were responsible in neuroprotection in neurotoxic rats. However, heartwood demonstrated better neuroprotective activities than the bark of P. marsupium due to some of the key compounds (1 R,4aR,7 R,8aR)-7-(2-Hydroxypropan-2-yl)-1,4a-dimethyldecahydronaphthalen-1-ol, 2-Propen-1-ol, 3-(2,6,6-trimethyl-1-cyclohexen-1-yl)-, 4-t-Butyl-2-[4-nitrophenyl]phenol, 1,1,4,7-Tetramethyldecahydro-1 H-cyclopropa[e]azulene-4,7-diol.

Cardio-protective and Antioxidant Potential of Pterocarpus marsupium heartwood extract on Isoproterenol induced heart failure in rats

Heart failure (HF) is a clinical syndrome characterised by a constellation of symptoms(dyspnea, orthopnea, lower limb swelling) and signs (elevated jugular venous pressure, pulmonary congestion) which have underlying cardiac abnormality resulting in reduced cardiac output and/or elevated intra-cardiac pressures. HF is a leading cause of mortality globally and despite major therapeutic advances, HF continues to cause substantial morbidity and mortality. Therefore, there is a need to develop new prophylactic and therapeutic strategies for HF. Isoproterenol (ISO) is a synthetic non-selective beta adrenoceptor agonist and is widely used to model toxic cardiomyopathy and heart failure. It induces myocyte damage mimicking myocardial infarction. In the present study, we used ISO to induce heart failure in Wistar rats and study the effect of Pterocarpus marsupium (PM) heartwood extract on it. PM is a well-recognised herbal drug known for its beneficial effect in diabetes with potential anti-inflammatory and anti-oxidative properties.Hypothesis: PM treatment will favourably modulate the cardiac hemodynamic, anti-oxidative stress and anti-inflammatory parameters compared to standard drug and control rats in isoproterenol induced heart failure rats.Methods & Results. Animals were randomly divided into 10 groups.1: Normal control rats fed normal pellet diet, 2: Normal control rats treated with Fluvastatin (10 mg/kg; oral gavage) for 15 days 3: Normal control rats treated with PM Extract (100 mg/kg; oral gavage) for 15 days. 4: Normal control rats subcutaneously injected with ISO {Heart Failure (HF) group} 5: HF rats treated with Fluvastatin (10 mg/kg; oral gavage) for 15 days. 6: HF rats treated with PM Extract (100 mg/kg; oral gavage) for 15days 7: HF rats treated with PM Extract (200mg/kg; oral gavage) for 15 days. 8: Rats pretreated with Fluvastatin (10 mg/kg; oral gavage) for 15 days and at the 15th day subcutaneously injected with ISO and continued Fluvastatin treatment for another 15 days 9: Rats pretreated with PM Extract (100 mg/kg; oral gavage) for 15 days and at the 15th day subcutaneously injected with ISO and continued PM Extract treatment for another 15 days 10: Rats pretreated with PM Extract (200 mg/kg; oral gavage) for 15 days and at the 15th day subcutaneously injected with ISO and continued PM treatment for another 15 days. The groups were compared pre and post treatment for cardiac hemodynamic parameters, anti- oxidative stress parameters (superoxide dismutase, nitric oxide, malondialdehyde nitric oxide), cardiac injury markers, anti-inflammatory parameters (Interleukin-6 and Tumor Necrosis Factor-α).The results of the present study indicate that prophylactic and therapeutic treatment PM heartwood extract improved the altered hemodynamic and biochemical parameters in isoproterenol-induced HF rats suggesting its cardio-protective action. None declared. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Letter to the editor on “in vitro & in silico study of hypoglycaemic potential of pterocarpus marsupium heartwood extract”

The Short Communication “In vitro & in silico study of hypoglycemic potential of Pterocarpus marsupium heartwood extract” published in Natural Product Research is praised for its depth of inquiry and exploration of hypoglycemic and antioxidant properties. However, there are areas for refinement, such as clearer definitions of key terms related to type 2 diabetes mellitus, updated literature review, improved organization and contextualization of the Results and Discussion section, and a more balanced discussion of the study’s implications.

A Polyherbal extract treats Streptozotocin induced Diabetes mellitus: Pharmacognostic study, HPLC analysis and in vivo antidiabetic activity

The plants used in study have been used traditionally for various purposes skin problems, diabetes, astringent, anti-inflammatory, antioxidant, hepatoprotective activities, anthelmintic, antipyretic, antidysentric, anti-hyperlipidemic, anticancer properties. The present study involves the evaluation of pharmacognostic parameters of the four herbal drugs (Azadirachta indica leaves, Pterocarpus marsupium heartwood, Picrorhiza kurroa rhizomes, and Withania coagulans berries and fruit coat), High performance Liquid chromatography (HPLC) as well as antidiabetic activity of polyherbal extract. The qualitative as well as quantitative analysis was done. The drug was given to Streptozotocin induced diabetic rats in doses 250 mg/kg and 500 mg/kg body weight for 21 days. In HPLC analysis it was found that major component in all the extracts was beta carotene along with hesperidin, tannic acid, catechin, ferulic acid, caffeic acid along with other polyphenolic components responsible for the antioxidant activity as well as antidiabetic activity of polyherbal extract. Rats were found to show an increase in body weight, improved blood sugar levels and hemoglobin. The liver glycogen, total proteins, serum lipid profile were restored back to normal range after using the polyherbal extract in diabetic rats. The histopathological study also shows an improvement in diabetic rats at cellular level. The results were found to be dose dependent.

In Silico Analysis of PTP1B Inhibitors and TLC-MS Bioautography-Based Identification of Free Radical Scavenging and α-Amylase Inhibitory Compounds from Heartwood Extract of Pterocarpus marsupium.

Type 2 diabetes mellitus leads to metabolic impairment caused by insulin resistance and hyperglycemia, giving rise to chronic diabetic complications and poor disease prognosis. The heartwood of Pterocarpus marsupium has been used in Ayurveda for a long time, and we sought to find the actual mechanism(s) driving its antidiabetic potential. Methanol was used to prepare the extract using a Soxhlet extraction, and the identification of metabolites was performed by thin-layer chromatography (TLC) and ultraperformance-liquid chromatography and mass spectroscopy (UP-LCMS). The antioxidant potential of methanolic heartwood extract of Pterocarpus marsupium MHPM was determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and a reducing power assay. The α-amylase and α-glucosidase enzyme inhibitory potential of MHPM were investigated for their antidiabetic activity against acarbose. TLC-MS-bioautography was performed to identify the compounds responsible for possible antioxidant and antidiabetic activities. Moreover, targeting protein tyrosine phosphatase 1B (PTP1B), a key regulator of insulin resistance, by identified metabolites from MHPM through molecular docking and all-atom molecular dynamics (MD) simulations was also undertaken, suggesting its potential as an antidiabetic herb. The IC50 of free-radical scavenging activity of MHPM against DPPH was 156.342 ± 10.70 μg/mL. Further, the IC50 values of MHPM in α-amylase and α-glucosidase enzymatic inhibitions were 158.663 ± 10.986 μg/mL and 180.21 ± 11.35 μg/mL, respectively. TLC-MS-bioautography identified four free radical scavenging metabolites, and vanillic acid identified by MS analysis showed both free radical scavenging activity and α-amylase inhibitory activity. Among the identified metabolites from MHPM, epicatechin showed significant PTP1B docking interactions, and its MD simulations revealed that PTP1B forms a stable protein-ligand complex with epicatechin throughout the progression, which indicates that epicatechin may be used as a promising scaffold in the development of the antidiabetic drug after isolation from Pterocarpus marsupium. Overall, these findings imply that Pterocarpus marsupium is a source of valuable metabolites that are accountable for its antioxidant and antidiabetic properties.

Quantitative Estimation of Phytoconstituents, Antioxidants and in vitro Antidiabetic Activity of Polyherbal Extract

Published on: October 2022Journal of Young Pharmacists, 2022; 14(4):382-386Original Article | doi:10.5530/jyp.2022.14.77 Authors:Navjot Kaur1,2,*, Shailesh Sharma31IKGPTU, Jalandhar, Punjab, INDIA.2Rayat Institute of Pharmacy, Railmajra, SBS Nagar, Rail, Punjab, INDIA.3ASBSJSM College of Pharmacy, Bela, Ropar, Punjab, INDIA. Abstract:Background: The herbal antidiabetic drugs are safe but effective alternate to conventional oral antidiabetic therapy. The single phytoconstituents of single plants are inadequate to produce the required efficacy therefore polyherbal can be tested for the desired results. The objective of the present study was the assessment of total phenolic, flavonoids, tannins content, in vitro antioxidant and antidiabetic activity of Azadirachta indica leaves, Picrorhiza kurroa rhizomes, Pterocarpus marsupium heartwood, and Withania coagulans berries and fruit coat. Materials and Methods: The ethanolic extract of all four drugs were prepared and mixed in a ratio of 1:1:1:1 to make the polyherbal extract. It was checked for the presence of phenols, flavonoids, and tannins. The High-Performance Liquid Chromatographic (HPLC) analysis for phenolic compounds as well as the beta carotenoids was done. The in vitro antioxidant activity as well as in vitro antidiabetic activity were calculated. Results: Quantitative estimations showed that maximum phenolic content was in Azadirachta indica and Pterocarpus marsupium, the flavonoid in Withania coagulans, and tannins content in Pterocarpus marsupium. The HPLC analysis showed the marked presence of beta carotenoids along with other phenolic compounds. The radical scavenging activity of Pterocarpus marsupium and the polyherbal extract was comparable to the standard Gallic acid. The total antioxidant capacity of polyherbal extract was found to be 15.22±.051 mg Quercetin equivalent (QE)/g dry plant extract (DPE). The polyherbal extract showed an IC50 value of 58.81 μg/ml in case of α- amylase inhibition and 64.81 μg/ml in case of α- glucosidase inhibition. Conclusion: The antioxidant activity, as well as antidiabetic activity, may be attributed to the phenolic, tannins, flavonoid content as well as beta carotene found in the polyherbal extract.Keywords: Azadirachta indica leaves, Pterocarpus marsupium heartwood, Picrorhiza kurroa rhizomes, and Withania coagulans berries and fruit coat, Antioxidant, In vitro antidiabetic.

Heartwood Extract of Pterocarpus marsupium Roxb. Offers Defense against Oxyradicals and Improves Glucose Uptake in HepG2 Cells.

Diabetes mellitus leads to cellular damage and causes apoptosis by oxidative stress. Heartwood extract of Pterocarpus marsupium has been used in Ayurveda to treat various diseases such as leprosy, diabetes, asthma, and bronchitis. In this study, we worked out the mechanism of the antidiabetic potential of methanolic heartwood extract of Pterocarpus marsupium (MPME). First, metabolic profiling of MPME was done using gas chromatography-mass spectrometry (GCMS), ultra-performance liquid chromatography-mass spectroscopy (UPLC-MS), and high-performance thin-layer chromatography (HPTLC) to identify phenols, flavonoids, and terpenoids in MPME. Biological studies were carried out in vitro using the HepG2 cell line. Many antidiabetic compounds were identified including Quercetin. Methanolic extract of MPME (23.43 µg/mL–93.75 µg/mL) was found to be safe and effective in reducing oxyradicals in HepG2 cells. A concentration of 93.75 µg/mL improved glucose uptake efficiently. A significant decrease in oxidative stress, cell damage, and apoptosis was found in MPME-treated HepG2 cells. The study suggests that the heartwood of Pterocarpus marsupium offers good defense in HepG2 cells against oxidative stress and improves glucose uptake. The results show the significant antidiabetic potential of MPME using a HepG2 cell model. The effect seems to occur by reducing oxidative stress and sensitizing the cells towards glucose uptake, hence lowering systemic glucose levels, as well as rescuing ROS generation.

Role of Pterostilbene in Metabolic Diseases through SIRT1 pathway- A Review

Pterostilbene (PTE) (3-5 dimethoxy-4-hydroxy-trans-stilbenes) is an analogue of resveratrol. It is extracted and isolated from a natural source of the heartwood of Pterocarpus marsupium Roxb., red grape skin, and blueberries (Vaccinium spp.). Substantial evidence suggested that PTE displayed numerous preventive and therapeutic properties in many metabolic disorders such as diabetes and obesity. Metabolic diseases result in Insulin resistance (IR) which advances to impaired sensitivity to insulin-mediated glucose disposal. The prominent role of SIRT (silent information regulator proteins) is now getting emphasized in metabolic disorders. SIRT1 represses Uncoupling protein 2 (UCP2) expressions which are further responsible for improving synthesis of ATP from glucose. This results in improving glucose utilization and insulin secretion, thus preventing IR. SIRT1 also exhibits prominent role in facilitating fatty acid mobilization thereby inhibiting adiposity. Metabolic disorders are therefore the consequences of SIRT1 downregulation. Pterostilbene, being a SIRT1 activator, increases insulin sensitivity reduces adiposity, therefore can prove to be beneficial in diabetes as well as obesity. The review summarizes therapeutic effects portrayed by Pterostilbene via the SIRT1 pathway in metabolic diseases.

Native Cyclodextrins as Complexation Agents for Pterostilbene: Complex Preparation and Characterization in Solution and in the Solid State

Pterostilbene (3,5-dimethoxy-4′-hydroxystilbene, PTB) is a natural dietary stilbene, occurring primarily in blueberries and Pterocarpus marsupium heartwood. The interest in this compound is related to its different biological and pharmacological properties, such as its antioxidant, anti-inflammatory, and anticarcinogenic activities and its capacity to reduce and regulate cholesterol and blood sugar levels. Nevertheless, its use in therapy is hindered by its low aqueous solubility; to overcome this limitation we studied the feasibility of the use of cyclodextrins (CDs) as solubility-enhancing agents. CDs are natural macrocyclic oligomers composed of α-d-glucose units linked by α-1,4 glycosidic bonds to form torus-shaped molecules, responsible for inclusion complex formation with organic molecules. In particular, the aim of this study was to evaluate the feasibility of complexation between PTB and native CDs using various preparative methods. The isolated solid products were characterized using differential scanning calorimetry (DSC), simultaneous thermogravimetric/DSC analysis (TGA/DSC), Fourier transform infrared (FT-IR) spectroscopy, and X-ray diffraction (XRD) on powder and single crystals. The results indicated little or no evidence of the affinity of PTB to complex with α-CD using the kneading method. However, with β-CD and γ-CD thermal analysis revealed an interaction which was also corroborated by FT-IR and 1H-NMR spectroscopy. With β-CD, a hydrated complex of PTB was isolated and its characterization by single-crystal XRD revealed, for the first time, the mode of inclusion of the PTB molecule in the cavity of a CD. To complement the solid-state data, liquid-phase studies were carried out to establish the effect of CDs on the aqueous solubility of PTB and to determine the complex stoichiometries and the association constants for complex formation. Phase-solubility studies showed AL-type profiles for α- and β-CD and a BS profile for γ-CD, with K1:1 values of 1144, 4950, and 133 M−1 for α-CD·PTB, β-CD·PTB, and γ-CD·PTB, respectively. The stoichiometry of CD·PTB complexes, determined by Job’s method, revealed for each system a 1:1 molar ratio. The dissolution rate of PTB was approximately doubled just by employing simple physical mixtures, but the best performance was achieved by products obtained via kneading and co-precipitation, which effected the complete dissolution of PTB in 40 and 20 min for β-CD and γ-CD, respectively.

ANTIDIABETIC ACTIVITY OF HEART WOOD OF PTEROCARPUS MARSUPIUM ROXB. IN ALLOXAN INDUCED DIABETIC RATS

Diabetes mellitus is most common disease which is leading cause of morbidity and mortality among Indian population. Pterocarpus marsupium is a medicinal plant used in ayurvedic medicine system to control blood sugar. The aim of this study was to determine antidiabetic effect of mixture of ethanolic extract of heartwood of Pterocarpus marsupium. To evaluate antidiabetic activity, extract was administered in experimental mice and significant decrease in blood glucose level was shown on 7th day. Histopathology was also studied. Thus from present study it can be concluded that pterocarpus marsupium extract has antidiabetic activity in alloxan induced diabetic rat.

Acute toxicity study and phytochemical screening of Pterocarpus marsupium heartwood

Background: Conventionally, Ayurvedic herbs are being used to treat various diseases. These medicinal compounds have to be evaluated for their safety and presence of therapeutic compounds for the clinical application. Aims and Objectives: The present study was designed to obtain the scientific knowledge on the safety profile as well as to assess the presence of pharmacologically active principles in the Pterocarpus marsupium heartwood. Materials and methods: The aqueous extract of P. marsupium heartwood was subjected to an acute toxicity in albino rats. The animals were divided into four groups (n = 6) and fed with graded doses (1000, 2000, and 5000 mg/kg p.o.) of plant extract, respectively, whereas control group had received 2 ml distilled water orally. Animals were continuously observed for the toxicological symptoms for 2 h and intermittently for 48 h and latter once in a day for 14 days. The body weight of the animals was recorded. In addition, the qualitative phytochemical investigations were conducted to identify the presence of active principles. Results: The animals fed with aqueous extract of P. marsupium heartwood did not exhibit any toxic symptoms and the mortality. However, there was a significant (P 5000 mg/kg. Furthermore, the phytochemical investigations of the plant extract showed the presence of carbohydrates, flavonoids, triterpenoids, saponins, tannins and phenols. Conclusion: The aqueous extract of P. marsupium heartwood was found to be safe and well tolerated even at a large dose of 5000 mg/kg. Furthermore, the plant extract found to possess pharmacologically active principles having wide pharmacological spectrum. Hence, it can be preferred in various therapeutic conditions.

Pterocarpus marsupium Roxburgh heartwood extract/chitosan nanoparticles loaded hydrogel as an innovative wound healing agent in the diabetic rat model

The application of modern nanomedicines to enhance wound healing is growing due to their simplicity for topical organization and fast flexibility with molecules that can boost and reinforce the process of healing even in patients with diabetes. The point of the present investigation was to synthesize effective Pterocarpus marsupium heartwood extract/chitosan nanoparticles (PM-CNPs) loaded hydrogel (PM-CNPsH) and evaluate its drug release efficiency, in-vitro anti-microbial activity and wound healing action in streptozotocin administered diabetic rat models. The prepared PM-CNPs further utilized for the preparation of various carbopol hydrogel formulations. Hence, prepared hydrogels are characterized for pH, morphology, consistency, and spreadability; thus, PM-CNPsH-1 is found to be the optimized formulation. In-vivo rats treated with PM-CNPsH-1 displayed significantly much quicker healing of wounds in diabetic and non-diabetic rats. The histological examination revealed re-epithelization and growth of granular tissue and an improvement in collagen deposition. These results indicate the effectiveness of optimized nanocomposites as a potential treatment for curing diabetic wounds.

Determining the Effect of Pterostilbene on Insulin Secretion Using Chemoproteomics.

Pterostilbene, the 3,5-dimethoxy derivative of resveratrol, is a well-known polyphenolic compound, mainly found in blueberries, grapevines, and Pterocarpus marsupium heartwood, which has recently attracted a great deal of attention due to its wide bio-pharmacological profile. Moreover, pterostilbene is more lipophilic than resveratrol, with a consequently better bioavailability and a more interesting therapeutic potential. In this work, a chemoproteomic approach, based on affinity chromatography, was applied on pterostilbene in the attempt to identify the biological targets responsible for its bioactivity. On this basis, syntaxins, a group of proteins involved in the formation of SNARE complexes mediating vesicles exocytosis, were selected among the most interesting pterostilbene interactors. In vitro and in cell assays gave evidence of the pterostilbene ability to reduce insulin secretion on glucose-stimulated pancreatic beta cells, opening the way to potential applications of pterostilbene as a supplement in the care of insulin-dependent metabolic disorders.

A Randomized Study to Determine the Sun Protection Factor of Natural Pterostilbene from Pterocarpus Marsupium

Ultraviolet (UV) rays and its harmful effects have always been a concern to skin health. Sunscreen and sunblock prevent the harmful effects of UV radiation on the skin. Sun Protection Factor (SPF) is an indication of the sun-protective capacity of an ingredient. There is an ever increasing interest in the cosmetic industry for developing novel functional ingredients from natural sources. The purpose of this study was to determine in-vitro and in vivo SPF of natural 90% pterostilbene extracted from the dried heartwood of Pterocarpus marsupium (Indian Kino). The SPF of purified pterostilbene and a formulation containing 0.4% pterostilbene was determined In Vitro using a UV spectrophotometer. Pterostilbene had an SPF of 21.73 ± 0.06, while the cream formulation had an SPF of 8.84 ± 0.01. The in vivo SPF of the 0.4% pterostilbene cream in humans was found to be 6.2 ± 1.30. Primary skin irritation tests in human subjects showed the formulation was safe and had no irritation potential. Pterostilbene was also found to have significant antioxidant activity as determined by free radical scavenging assays in vitro. These results suggest that natural pterostilbene is an antioxidant and shows SPF value both in-vitro and in the human clinical study and thus could be used as an ingredient in topical sun-protective formulations.

Inhibitory Activity of Asana, Heartwood of Pterocarpus marsupium, Against Xanthine Oxidase

The heartwood of Pterocarpus marsupium is called as “Asana” in Ayurveda. Its aquatic infusion was used for treating “prameha,” which indicates a polyuria disease in modern medicine. In our research program to investigate a novel agent to improve hyperuricemia, we focused on the extract of Asana as a xanthine oxidase (XOD) inhibitor. Asana extract (50% ethanolic extract, PM-ext) showed 11%, 35%, and 38% inhibition at 50, 200, and 500 µg/mL, respectively. Subsequently, PM-ext was partitioned with ethyl acetate (AcOEt), butanol, and water. Among them, AcOEt-soluble fraction indicated the most potent XOD inhibitory activity and was consecutively fractionated using various liquid chromatography to obtain liquiritigenin (1), isoliquiritigenin (2), and marsupsin (3) as active principles. Compound 1 showed 16% inhibition at 200 µM while 2 showed 20%, 32%, and 46% inhibition at 50, 100, and 200 µM, respectively. Compound 3 showed 15% inhibition at 500 µM. This is the first report on the XOD inhibitory activity of 3. From these results, PM-ext is a promising candidate material for improvement of hyperuricemia. Here, Asana was recognized as an effective material against noncommunicable disease and is expected to be developed as a functional ingredient.

Anti-tyrosinase and Anti-oxidative Activities by Asana: the Heartwood ofPterocarpusmarsupium

Asana (the heartwood of Pterocarpus marsupium) has been utilized as an agent for diabetes mellitus in Ayurveda traditional medicine. In our research program to explore novel functions of asana extract, we focused on its skin-whitening effect because asana has been used as a remedy for chronic skin diseases. In addition, the authors have already reported an improvement in blood fluidity that brightens dull facial skin. Based on these effects, asana is a promising candidate agent that possesses both blood fluidity and anti-tyrosinase activities. We focused on the anti-tyrosinase activity and anti-oxidative activities of asana and the results are summarized in this report. We found that a 50% ethanolic extract obtained from asana (PM-ext) showed 23%, 53%, and 71% inhibition against mushroom tyrosinase at 12.5, 50, and 200 µg/mL. Oxyresveratrol and isoliquiritigenin were identified as the active compounds by activity-guided purification. Oxyresveratrol has higher potency than isoliquiritigenin and the IC50of oxyresveratrol was estimated to be 2.1 µM. On the other hand, isoliquiritigenin showed 21%, 28%, and 38% inhibition at 10, 50, and 100 µM, respectively. The inhibitory activity of oxyresveratrol was compared with 3 stilbenes, pterostilbene, resveratrol, and piceatannol. Although oxyresveratrol showed 72.8%, 81.0%, and 85.4% inhibition at 2, 5, and 10 µM, respectively, pterostilbene, resveratrol, and piceatannol showed no effects at the same concentration; these compounds also demonstrated anti-melanogenesis activity on B16 murine melanoma cells. As a result, oxyresveratrol showed the most potent activity, without cytotoxicity, with 38%, 74%, and 84% inhibition at 2, 10, and 20 µM, respectively, while pterostilbene showed 26%, 71%, and 79% inhibition at the same concentration with cytotoxicity at 10 and 20 µM. Resveratrol showed 20%, 41%, and 57% inhibition without cytotoxicity at 2, 10, and 20 µM, respectively. Auto-oxidation is one of the major factors in melanin biosynthesis and anti-oxidative activity is suitable for an anti-melanogenesis agent. We investigated the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity by PM-ext. As a result, PM-ext showed 16%, 33%, and 73% DPPH radical-scavenging activity at 10, 20, and 50 µg/mL, respectively. Oxyresveratrol showed 19%, 31%, and 59% scavenging activity at 10, 20, and 50 µM, respectively, similar to piceatannol. In addition, PM-ext showed 29%, 48%, and 80% suppressive activity on AGEs production at 3.1, 12.5, and 50 µg/mL, respectively. Oxyresveratrol showed 32%, 47%, and 55% activity at 10, 50, and 100 µM, respectively, and this was the most potent among the stilbenes tested. These results suggest that PM-ext could be a promising candidate as skin-whitening agent.

Anti-diarrhoeal activity of ethanolic extract of heartwood of Pterocarpus marsupium roxb.

Diarrhoea is a common cause of death in developing countries and second most common cause of infant’s death worldwide. Pterocarpus marsupium is a medicinal herb belonging to the family Fabaceae has been traditionally used in the treatment of diarrhoea. They were found to contain tannins, alkaloids, saponins, sterols, triterpenes and reducing sugars. This study evaluated the antidiarrhoeal activity of ethanolic extract of heartwood of Pterocarpus marsupium induced by castor oil and magnesium sulphate in rat at 200 and 400 mg/kg b.w. The doses were given orally and showed significant antidiarrhoeal activity comparable with that of the standard drug loperamide. The statistical analyses of results were carried out using one-way analysis (ANOVA) followed by Student t-test. On the basis of these findings, it can be assumed that Pterocarpus marsupium could be a potential source for novel discovery for antidiarrhoeal. These results may support the fact that this plant is used traditionally to cure diarrhoea.
 Keywords: Pterocarpus marsupium, Anti-Diarrhoeal, Castor Oil, Magnesium Sulphate, Loperamide

Identification and quantification of secondary metabolites of Pterocarpus marsupium by LC–MS techniques and its in-vitro lipid lowering activity

Pterocarpus marsupium (Vijaysar) is a deciduous tree, considered to have high insulinogenic properties. Since ancient times Ayurveda practitioners believed that heartwood of vijaysar significantly reduced blood sugar levels in diabetes. Marsupsin, pterosupin and pterostilbene are the three important phenolic constituents of the heartwood of this plant reported to significantly lower the blood glucose level. For the dereplication of bioactive phytochemicals present in heartwood of Pterocarpus marsupium a method was developed to identify and elucidate the main fragmentation pathways of phytoconstituents in positive electrospray ionization mode using HPLC-ESI-QTOF-MS/MS. Further, an UPLC-ESI-QqQLIT-MS/MS method was developed to determine the contents of bioactive compounds, namely 2,6-dihydroxyphenyl glucopyranoside, pterosupol, vijyoside, marsuposide, pteroside, pterocarposide, epicatechin, vanillic acid, quercetin, formononetin and naringenin in heartwood of this plant. Twenty seven compounds were tentatively identified based on their retention time, accurate mass and MS/MS patterns. In-vitro assessment of compounds in 3T3-L1 adipocytes revealed significant attenuation of lipid accumulation in extract from Pterocarpus marsupium, hence it may have certain phytochemicals which are responsible for reducing obesity. The analytical method for quantitation was validated and successfully applied for the simultaneous determination of eleven bioactive compounds. In conclusion, rapid and accurate HPLC-ESI-QTOF-MS/MS and UPLC-ESI-QqQLIT-MS/MS methods developed in positive ionization mode were successfully applied for identification and quantitation of bioactive phytochemicals present in heartwood of Pterocarpus marsupium.

In vitro & in silico study of hypoglycemic potential of Pterocarpus marsupium heartwood extract

Total amount of phenols, flavonoids and tannins in methanolic extract of heartwood Pterocarpus marsupium were found to be 124.4 ± 4.6, 485.5 ± 2.7, and 71.6 ± 4.2 mg/gm of plant extract respectively. HPLC analyses of extract showed retention time at 6.2 min and FTIR analysis confirmed presence of Liquiritigenin in methanolic extract. In DPPH scavenging activity and ABTS∙+ scavenging activity, IC50 values were found to be 138.3, 12.4, 13.5 and 47.8, 3.9, 4.2 μg/mL for aqueous, methanolic extract and standard ascorbic acid respectively. In α-amylase inhibition assay and α-glucosidase inhibition assay, IC50 values for standard Acarbose, aqueous and methanolic extract of heartwood were 44.09, 166.72, 48.20 and 45.17, 172.32, 48.12 respectively. Molecular docking study showed hydrogen bonding between Liquiritigenin and catalytic triad (Asp197, Glu233, and Asp300) of α-amylase and His407 of α –Glucosidase with −5.60 and −7.10 binding energies respectively.

Evaluation of effect of alcoholic extract of heartwood of Pterocarpus marsupium on in vitro antioxidant, anti-glycation, sorbitol accumulation and inhibition of aldose reductase activity

Rising popularity of phytomedicines in various diseased conditions have strengthened the significance of plant-research and evaluation of phytoextracts in clinical manifestations. Pterocarpus marsupium Roxb., a medicinal plant, known for its anti-oxidant and anti-diabetic activity is a rich source of phytochemicals with antihyperglycemic and antihyperlipidemic activities. However, its possible role in diabetic complications is not evaluated yet. The present study explores the possible role of alcoholic extract of heartwood of P. marsupium in the treatment of long-term diabetic complications. The alcoholic extract of P. marsupium was evaluated for advanced glycation-end-products formation, erythrocyte sorbitol accumulation and rat kidney aldose reductase enzyme inhibition at the concentration of 25–400 μg/ml using in-vitro bioassays. Also the phytoextract at the concentration of 10–320 μg/ml was evaluated for its antioxidant potential by in-vitro antioxidant assays which includes, determination of total phenol content; reducing power assay; nitric oxide scavenging activity; superoxide radical scavenging activity; total antioxidant capacity; total flavonoid content; DPPH scavenging activity; and hydrogen peroxide scavenging activity.The alcoholic extract of P. marsupium across varying concentrations showed inhibitory effect as evident by IC50 on advanced glycation-end-products formation (55.39 μg/ml), sorbitol accumulation (151.00 μg/ml) and rat kidney aldose reductase (195.88 μg/ml). The phytoextract also exhibited high phenolic and flavonoid contents with promising antioxidant potential against the antioxidant assays evaluated. The present investigation suggests that the phytoextract showed prominent antioxidant, antiglycation property and, inhibited accumulation of sorbitol and ALR enzyme, thus promising a beneficial role in reducing/delaying diabetic complications.