In Vitro Evaluation of Selected Fruit Extracts Against Cardiovascular Diseases
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In vitro Evaluation of Selected Fruit Extracts against Cardiovascular Diseases Adapa Satish Kumar1, P. Dwarakanadha Reddy2, S. V. Satyanarayana3 1Research Scholar, Jawaharlal Nehru Technological University Anantapur, Anantapuramu, Andhra Pradesh, India, 2Department of Pharmaceutics, Annamacharya College of Pharmacy, Rajampet, Andhra Pradesh, India, 3Department of Chemical Engineering, JNTUA College of Engineering, Ananthapuramu, Andhra Pradesh, India. Abstract Objective: Cardiovascular diseases (CVDs) are the number one cause of death globally, more people die annually from CVDs than from any other cause. Much of this interest centers on the use of antioxidant vitamins and the antioxidant properties of herbal materials, although some herbal materials may also improve ORIGINAL ARTICLE ORIGINAL conventional cardiovascular risk factors or have antithrombotic effects. This study was undertaken to investigate the in vitro evaluation of selected fruit extracts against cardiovascular metabolic syndrome with methanolic extracts of Schisandra (Magnolia vines), Muntingia calabura, and Alangium salviifolium fruits. Materials and Methods: Angiotensin-converting enzyme (ACE) inhibition assay, assay of nitric oxide (NO) scavenging activity, and in vitro α-amylase inhibitory studies were carried out to evaluate the cardiovascular metabolic syndrome activity of methanolic extract of Schisandra (M. vines), M. calabura, and A. salviifolium fruits. Results: The preliminary phytochemical screening showed the presence of various phytoconstituents such as flavonoids, phenolic compounds, triterpenoids, tannins, saponins, amino acids, proteins, and carbohydrates in the fruit extracts. The ACE inhibitory activity of fruit extracts was represented as percentage ACE inhibition. The fruits extract demonstrated ACE inhibitory activity at a concentration of 800 µg/ml, showing an inhibition >50%. Statistically significant results were observed in in vitro α-amylase inhibitory assay and in NO scavenging assay. Conclusion: The role of redox mechanisms in the control of expression and activity of rennin-angiotensin system (RAS) enzymes and angiotensin receptors may provide important insight into the function of local tissue RAS in health and disease states. The selected fruit extracts have promising role against CVDs. Key words: Alangium salviifolium, cardiovascular metabolic syndrome activity, Magnolia vines, Muntingia calabura INTRODUCTION for population-based strategies for CVD risk reduction. Epidemiological and clinical data: Epidemiological and he cardiovascular disease (CVD) is clinical studies indicate that the risk of CVD is reduced common, morbid, and responsible for by a diet rich in fruits, vegetables, unrefined grains, fish, about 17.3 million deaths annually and low-fat dairy products, and foods low in saturated fats T [1] and sodium are helpful. Other foods such as mono- and worldwide. The modifiable risk factors for CVD include obesity, hypertension, polyunsaturated fats, brans, nuts, plant sterols, and soy hyperlipidemia, type 2 diabetes mellitus, MetS, proteins have all been shown to have a favorable effect on and lifestyle risk factors such as smoking, lipid profile and blood pressure, and the overall CV health. physical inactivity, and dietary factors.[1,2] The Foods and nutrients play a key role in functioning of various nutritional factors have an important bearing Address for correspondence: on the cardiovascular (CV) health, either Dr. P. Dwarakanadha Reddy, directly or through their effects on various Department of Pharmaceutics, Annamacharya College of CV risk factors including hypertension, Pharmacy, Rajampet, Andhra Pradesh, India. dyslipidemia, and diabetes mellitus. The E-mail: [email protected] protective effects against CVD have been demonstrated for various nutraceuticals and Received: 28-11-2020 dietary supplements,[3] and these simple Revised: 21-01-2021 lifestyle interventions open practical, Accepted: 06-02-2021 potentially easy, and affordable possibilities Asian Journal of Pharmaceutics • Jan-Mar 2021 • 15 (1) | 94 Kumar, et al.: Fruit extracts activity against cardiovascular diseasea body organs and are helpful in maintaining health and Muntingia is a genus of plants in the family Muntingiaceae, reducing the risk of various diseases. comprising only one species, Muntingia calabura. It is native to the neotropics, from Mexico South to Bolivia. M. calabura The nutraceutical is defined as “a food component that provides is a shrub or tree up to 12 m tall with spreading branches. The potential medical or health benefits, including the prevention leaves are alternate, distichous, oblong or lanceolate, 4–15 cm and treatment of disease.”[4] The definition includes medicinal long and 1–6 cm wide, with toothed margin and covered in products made from natural ingredients. The nutraceuticals short hairs. The flowers are small (up to 3 cm wide), solitary supplement the diet and also aid in the prevention and/or or in inflorescences of 2–3 flowers; with 5 lanceolate sepals, treatment of disease. Early research evaluated the benefits of hairy; 5 obovate white petals; many stamens with yellow plant-derived foods based on their Vitamin C, Vitamin E, and anthers and a smooth ovoid ovary. Fruit an edible berry, red carotenoid content. More recent work pointed out correlation at maturity, about 1.5 cm wide.[10] of benefits with individual compounds. However, the effects noted by testing them alone may be related to the synergistic action of the myriad of other bioactive components present MATERIALS AND METHODS in foods. In each family of bioactive compounds, there are usually various members present. Plant material and preparation of extract Rennin-angiotensin system plays an important role in the Schisandra (M. vines), M. calabura, and Alangium normal control of CV and renal function, but the system salviifolium fruits were obtained from the local places of also becomes a contributing factor in the development and Tirupati, AP. The plant was authenticated by Dr. K. Madhava progression of various types of CVD, including hypertension Chetty M.Sc., M.Ed., M.Phil., Ph.D., PG DPD., Department and heart failure. The links between angiotensin (Ang) of Botany, SVU, Tirupati, AP. pathways and nitric oxide (NO) production and signaling effects centers on Ang-(1–7) and activation of its specific Fruits without rind were treated with sufficient amount receptor Mas, a G-protein coupled receptor encoded by the of pure methanol for 1 week at room temperature with Mas protooncogene. Mas receptor stimulation activates PI3K- occasional shaking. The extract was filtered through a cotton dependent Akt phosphorylation, leading to NO production by plug followed by Whatman No. 1 filter paper. The filtrate was phosphorylation of endothelial NO synthase.[5] Mas activation then evaporated under reduced pressure to give a dark green can also lead to NO production through neural NO synthase,[6] viscous mass and stored at 4°C until use. although the intermediate pathways are not yet defined. Ang-(1– 7)/Mas receptor activation can also inhibit mitogen-activated protein kinase phosphorylation, stimulate phospholipase Phytochemical evaluation A2 activity and consequently release of arachidonic acid for The methanolic extraction of M. vines (MEMV), methanolic prostanoid production, and interfere in various ways with Ang extraction of M. calabura (MEMC), and methanolic II/AT1 receptor mediate signaling pathways.[7] extraction of A. salviifolium (MEAS) were screened for the presence of various phytoconstituents such as carbohydrates, Sage-leaved alangium is a small, bushy tree offering a dense proteins, flavonoids, polyphenolic compounds, saponins, canopy with a short trunk. It bears fragrant white flowers tannins, and triterpenoids.[11] which have green buds. The petals of the flower typically curl backwards exposing the multiple stamens and a linear stigma distinctly sticking out. The fruits are spherical berry Ang-converting enzyme inhibition assay like and red and the prominent remains of the calyx can be seen distinctly with a white color. The leaves are simple, The Ang-converting enzyme inhibitory activity was carried alternate, and oblong-lanceolate and stunted branches end up out using (N-[3-(2-furyl) acryloyl]-Phe-Gly-Gly) (FAPGG) as with sharp ends making it look like thorns.[8] the substrate.[12] The extracts and the standard drug lisinopril (1 mg/ml) were prepared by dissolving in reaction buffer Schisandra (Magnolia vines) is a genus of twining shrub that (HEPES 25 mM, NaCl 293 mM, pH 8.3). The assay mixture generally climbs on other vegetation. Some early publications (750 µl) consisting of 530 µl of FAPGG (3 mM in reaction misspell the name as “Schizandra,” but “Schisandra” is correct. buffer) and 200 µl of extract at different concentrations (100– Various authors have included the plants in the Illiciaceae 800 µg/ml) was incubated for 3 min at 37°C. The reaction was Schisandra is native to Asia and North America, with a center initiated by adding 20 µl of Ang-converting enzyme (ACE) of diversity in China. Some species are commonly grown in solution (0.05 U/ml) to the test reaction and the samples were gardens as ornamentals. It is a hardy deciduous climber which incubated for 1 h at 37°C. The reaction was then stopped by thrives in almost any kind of soil; its preferred position is on adding 80 µl of 5% trifluoroacetic acid solution and samples a sheltered, shady wall. It may be propagated by cutting off were centrifuged at 9000 rpm for