PHARMACOGNOSTIC ANALYSIS OF DIOSCOREA BULBIFERA L.: A GATEWAY TO ITS MEDICINAL BENEFITS

PHARMACOGNOSTIC ANALYSIS OF DIOSCOREA BULBIFERA L.: A GATEWAY TO ITS MEDICINAL BENEFITS

Yuvraj Vishwakarma *, Vikas Pandey, Rajesh Shukla, Devendra Dhanorya, Gourav Bairagi and Vandana Gupta

Faculty of Pharmaceutical Science, Mangalayatan University, Jabalpur, Madhya Pradesh, India.

ABSTRACT: Medicinal plants contain phytochemicals that are recognized for their wide range of therapeutic uses. Dioscorea bulbifera often referred to as the air potato or bitter yam, is a plant belonging to the Dioscoreaceae family, which includes about 600 species. It holds significant traditional medicinal importance worldwide, being used for treating various ailments. This plant is commonly found in India, Maldives, China, Japan, Indonesia, and is a staple food crop in West Africa. The plant features bulbous aerial tubers, toxic substances like the alkaloid dioscorine, and small, dioecious flowers. Dioscorea bulbifera grows in tropical and subtropical regions, preferring loamy soils and temperatures between 20°C and 30°C. It reproduces mainly asexually via bulbs. Dioscorea bulbifera is widely used in traditional Indian and Chinese medicine for conditions such as piles, dysentery, syphilis, ulcers, pain relief, and inflammation. The plant's tubers and bulbils are noted for their cytotoxic properties, traditionally used in treating sore throats, goitre, and various other conditions. Phytochemical analyses reveal that the plant contains alkaloids, glycosides, sterols, polyphenols, flavonoids, and saponins, contributing to its medicinal uses. The presence of these compounds varies with the plant's geographical origin, highlighting its versatile medicinal potential.

Keywords: Dioscorea bulbifera, Antihyperlipidemic, Anti-microbial, Anti-HIV-IN, Anti- tumour, Anti-funga

INTRODUCTION: Phytochemicals found in medicinal plants are abundant and have a wide range of therapeutic applications ¹. Dioscorea bulbifera is a distinctive medicinal plant within the Dioscoreaceae family, which comprises around 600 species. It holds significant importance in traditional medicine worldwide ². The common names for Dioscorea bulbifera include air potato, air yam, bitter yam, cheeky yam and air potato. These names reflect its bulbous aerial tubers and its distribution in various regions ³. Dioscorea bulbifera is typically found in Indonesia, Japan, China, Maldives, and India. It serves as a staple food crop in West Africa.

These plants are climbing, perennial, erect herbs, or shrubs with woody stems reaching about 10 meters in length. The leaves are simple, opposite, lobed, and alternate, measuring approximately 20 cm in length with a reticulate venation pattern. The stems are herbaceous and emerge from underground tubers. The aerial bulbs contain toxic substances and the alkaloid dioscorine. Flowers are generally small, minute, dioecious, and occasionally bisexual ⁴. Dioscorea bulbifera is commonly grown in tropical and subtropical regions with moist conditions. It thrives in temperatures ranging from 12°C to 38°C, with optimal growth occurring between 20°C and 30°C.

The plant prefers loamy soil enriched with high organic content for cultivation ⁵. Sexual reproduction in Dioscorea bulbifera is uncommon and typically occurs through seeds, while asexual reproduction primarily occurs via bulbs ⁶. This plant is frequently utilized in traditional Indian and Chinese medicinal practices ⁷. Dioscorea bulbifera L. var. Sativa is employed in Bangladesh for treating leprosy and tumors ⁸. In the western highlands of Cameroon, native people use Dioscorea bulbifera for treating pig cysticercosis, although the tubers are often discarded after harvest because of their intense bitterness. The roots of Dioscorea bulbifera, valued for their cytotoxic properties, have traditionally been used in Chinese medicine to address sore throats and goiter ⁹. In India, the bulbs of Dioscorea bulbifera are used to treat ailments like piles, dysentery, and syphilis and are applied externally for ulcers, pain relief, and to reduce inflammation ¹⁰. In traditional Chinese medicine, Dioscorea bulbifera has been widely used by practitioners to treat various conditions, including hemoptysis, goiter, skin infections, orchitis, pharyngitis, and cancer ¹¹. Additionally, Dioscorea bulbifera has been utilized for its blood-clotting properties to stop bleeding and for detoxifying poisons. Phytochemical studies have revealed compounds in the tubers that are associated with a range of medicinal uses ⁴⁴.

Occurrence and Distribution: Dioscorea bulbifera, also called air potato or air yam, is native to tropical regions of Asia, northern Australia, the Americas, and sub-Saharan Africa. In China, it grows in the southern provinces, including Anhui, Fujian, Gansu, Guangxi, Guangdong, Guizhou, Henan, Jiangsu, Tibet, and Yunnan. Known in Chinese herbal medicine as "Huangdu" and "Huangyaozi," this plant has traditional significance in the region ^{11, 12}.

Flowers: From September to November.

Fruit: Starting from December.

Part Used: Roots, Bulbils, Tubers ¹³.

Taxonomic Classification:

TABLE 1: TAXONOMIC CLASSIFICATION ¹³

Vernacular Names:

TABLE 2: VERNACULAR NAMES ¹⁴

MORPHOLOGY: Dioscorea bulbifera, commonly known as air potato, is a vigorously twining perennial vine that can grow up to 20 meters or more in length. Its stems are non-spiny and freely branching above, with round or slightly angled internodes that twine counter-clockwise. The plant possesses two types of storage organs: aerial bulbils located in the leaf axils of the climbing stems and underground tubers. The tubers resemble small, oblong potatoes and have a bitter taste. The bulbils, which are pale and round to globose, can be up to 13 cm wide and are a distinguishing feature of the plant. The leaves of D. bulbifera are visually appealing, arranged alternately, and have a broad heart shape, with long petioles connecting them to the stem. They measure 10-15 cm in length and 7.5-10 cm in width, with a deeply cordate base and an acuminate to shortly caudate apex. The leaves are membranous, glabrous, and have 9-11 prominent, radiating veins originating from the point where the petiole attaches. The petioles can be up to 20 cm long. Flowers are rare in D. bulbifera. When present, they are small, pale green, and fragrant, arising from the leaf axils. Male flowers appear in slender, axillary panicled spikes that are pendulous and can reach up to 18 cm in length. The bracteoles are ovate and acute ¹⁵.

The perianth of Dioscorea bulbifera is light green, with six biseriate lobes, each about 2.5 mm long and linear-oblong in shape. The plant has six free stamens. Female spikes appear in clusters of 1-3, with three staminodes. The ovary is triquetrous and 3-locular, containing two ovules per locule. The styles are three in number, with a 2-fid, reflexed stigma. The capsules, which serve as the fruit, measure 1.5-2.3 cm by 1-1.5 cm, are oblong, and have three wings. The seeds are partially winged. Dioscorea bulbifera reproduces both sexually, through seeds, and vegetatively, through underground and aerial tubers (bulbils).

This dual mode of reproduction allows it to spread quickly and dominate entire forests within a single growing season. The aerial stems die back in the winter, but the plant resprouts from the bulbils and underground tubers ¹³.

FIG. 1: DISCOREA BULBIFERA: (I) LEAF (II) TUBER (III) WHOLE PLANT ¹⁷ (IV) STEM ¹⁸

The bulbil of Dioscorea bulbifera is fairly hard and heavy, dish-shaped, measuring up to 12 cm (5 inches) by 10 cm (4 inches). It is brown with numerous, uniformly distributed, tubercle-like eyes. The plant produces abundant bulbils of various sizes and shapes; in some cultivars, the tuber is suppressed in favour of larger bulbils that contain all the reserve food. Typically, small bulbils are warted, but larger ones can be smooth. Tubers are typically small and round but can reach larger sizes when cultivated, occasionally weighing as much as 1 kg. The edibility of the tubers varies by variety; they can be either toxic or edible. Tubers are renewed annually and have a purplish-black or earth-coloured skin, often coated with numerous small feeding roots, although some cultivated varieties have smooth skin. The flesh inside can range from white to lemon yellow, sometimes with purple flecks, and is very mucilaginous Fig. 1. The tubers have a few root and root scars, with a dark brown outer surface and a yellow to light brown inner surface. The odour is indistinct, and the taste is bitter ¹³.

Phytoconstituents: Phytochemical research on Dioscorea bulbifera has revealed that it contains a variety of compounds, including alkaloids, glycosides, proteins, fats, sterols, polyphenols, tannins, flavonoids, and saponins, with their occurrence possibly varying depending on the country where the plant is sourced. Inorganic micronutrients found in Dioscorea bulbifera include iron (Fe), copper (Cu), zinc (Zn), manganese (Mn), cobalt (Co), molybdenum (Mo), vanadium (V), boron (B), chlorine (Cl), iodine (I), bromine (Br), and sodium (Na) ¹⁹.

TABLE 3: PHYTOCONSTITUENTS OF DIOSCOREA BULBIFERA

Chemical Structures of Several Key Phytochemicals Present in Dioscorea bulbifera:

TABLE 4: ETHNOBOTANICAL APPLICATIONS OF DIOSCOREA BULBIFERA

Pharmacological Activities:

Antibacterial Activity: The air-dried bulbils of Dioscorea bulbifera were finely ground and extracted three times (for 72 hours each) with hexane, dichloromethane, ethyl acetate, methanol, ethanol, and distilled water. The resulting extract was filtered using Whatman No. 1 filter paper under vacuum and concentrated by evaporating the solvent with a rotary evaporator. The extract was subsequently freeze-dried and kept at -20 °C until required for use. To assess the antimicrobial properties of Dioscorea bulbifera extracts, common skin pathogens were selected. The bacterial strains tested included Staphylococcus aureus DMST 8840, methicillin-resistant Staphylococcus aureus (MRSA) DMST 20651, Staphylococcus epidermidis DMST 3547, Staphylococcus epidermidis DMST 4343, and Pseudomonas aeruginosa DMST 4739.The bacterial strains were subcultured on brain heart infusion agar (BHA) and incubated at 37 °C for 24 hours before testing ⁴⁸. To prepare the extract, 64 mg/mL was dissolved in 10% DMSO and subsequently diluted in Mueller Hinton broth (MHB) to obtain concentrations ranging from 0.006 to 64 mg/mL in a 96-well microtiter plate. A 10% DMSO solution served as the control. Fifty microliters of the extract and control were used in the analysis. The bacterial strains were adjusted to a turbidity equivalent to a 0.5 McFarland standard and then diluted 1:100 in MHB. Fifty microliters of this bacterial suspension were added to each well. The microtiter plates were gently tapped at the corners to ensure thorough mixing. Tetracycline served as a positive control, and the plates were incubated at 37 °C for 24 hours. Each minimum inhibitory concentration (MIC) determination was conducted in triplicate, with MIC defined as the lowest concentration of the extract that prevented visible bacterial growth ⁴⁸. As represent in Table 5. The minimum inhibitory concentration (MIC) was used to evaluate the inhibitory effects of the crude ethyl acetate extract and the antibacterial fraction from Dioscorea bulbifera on all tested bacteria. The MIC values ranged from 0.78 to 1.56 mg/mL for the ethyl acetate extract and from 0.02 to 0.78 mg/mL for the antibacterial fraction. The antibacterial fraction showed the strongest effect against Staphylococcus epidermidis 3547. The ethyl acetate extract of Dioscorea bulbifera demonstrated significant antibacterial activity, as reflected in its MIC values. The antibacterial fraction exhibited notably stronger activity against the bacterial strains tested, with lower MIC values across all species, suggesting a higher concentration of active components within this fraction that enhances its antimicrobial effect ⁴⁸.

TABLE 5: MINIMUM INHIBITORY CONCENTRATIONS (MIC) OF ETHYL ACETATE EXTRACT AND FLAVANTHRIN-CONTANING FRACTION ⁴⁸

Antifungal Activity: Extracts from the leaves, stems, and tubers of Dioscorea bulbifera were screened for antifungal activity against Candida parapsilosis (MTCC 2513). The antifungal activity was assessed using a slightly modified version of the standard disc diffusion method ^{49, 50}.

Each extract was dissolved in dimethyl sulfoxide (DMSO), and three concentrations of the plant extracts (25, 50, and 100 mg/mL) along with standard drugs were prepared. Discs measuring 6 mm in diameter were soaked in the solutions for 12 hours before being placed on swabbed petri plates. After incubating the plates at 37°C for 18 to 24 hours, the zones of growth inhibition around the discs were measured. The sensitivity of the microbial species to the plant extracts was evaluated by measuring the size of the inhibitory zones, including the diameter of the discs. Zones measuring less than 8 mm were considered inactive against the microorganisms ⁵¹. The media used for antifungal test was Sabouraud’s dextrose agar/ broth of Hi media Pvt. Bombay, India. It was noted that the peels of Dioscorea bulbifera tubers are traditionally used to treat fungal skin infections. Tribal communities in Odisha and Jharkhand collect these tubers from the wild, dry them, peel off the upper layer, and macerate it. The resulting paste is applied externally to treat fungal skin infections. To validate this traditional use, antifungal activity tests were conducted, and the results indicated that tuber extracts were more effective than leaf and stem extracts.

TABLE 6: ANTIFUNGAL ACTIVITY (ZONE OF INHIBITION INMM, MEAN± SD; N=3) OF DIOSCOREA BULBIFERA LEAF, STEM, TUBER) AGAINST MTCC 2513 ⁵¹

(mm: millimetre; SD: Standard Deviation; n: replica; ≤: less than or equal to).

Antihyperlipidemic Activity: The aerial tubers of Dioscorea bulbifera were freshly harvested from a farm in Amiri, Imo State, Nigeria. They were sliced into smaller pieces and shade-dried for two weeks. The dried tubers were ground into a coarse powder and then exhaustively extracted using a Soxhlet apparatus with a hydro methanol mixture (80:20). The extraction and subsequent phytochemical screening were conducted following the methods described by Odebiyi and Sofowora ⁵².

The extracts were filtered and then concentrated to dryness using a rotary evaporator at 40-50°C. The hydromethanol extracts were subsequently stored at 2-5°C until needed for further use. Fifty-five adults male Wistar rats, weighing between 180 and 250 grams, were obtained from the Animal House of the Department of Human Physiology at the University of Port Harcourt for the study. The rats underwent a three-week acclimatization period before the study began, kept under standard laboratory conditions: temperature between 25-29°C, 55-65% relative humidity, and a natural light/dark cycle. During acclimatization, the rats were fed balanced commercial rat chow (Top Feed LTD., Sapele, Nigeria) ad libitum. Hyperlipidaemia was naturally induced in the rats by feeding them a specially formulated high-fat diet, consisting of 80% commercial rat chow (Top Feed LTD., Sapele, Nigeria) and 20% rendered cow fat. The experimental design involved dividing the male Wistar rats into eleven groups, consisting of five control groups and six experimental groups. The lipid profile, serum glucose levels, and atherogenic index of Wistar rats, which were induced with hyperlipidaemia using a high-fat diet, tyloxapol, and dexamethasone, were assessed after treatment with the hydromethanolic extract of Dioscorea bulbifera ⁵³.

Anti-HIV-1 IN Activity: The bulbils of Dioscorea bulbifera were harvested in 2011 from Uttaradit Province, Thailand. A botanist from the Forest Herbarium, Department of National Parks, Wildlife and Plant Conservation, Thailand, identified the plant. The voucher specimen (SKP 062040201) has been stored at the Faculty of Pharmaceutical Sciences; Prince of Songkla University ⁵⁵. The HIV-1 integrase (IN) protein was generously supplied by Dr. Robert Craigie from the National Institutes of Health, Bethesda, MD. This enzyme was produced in Escherichia coli and purified following the method outlined by ⁵⁴. Dried bulbil powders of Dioscorea bulbifera (1.7 kg) were macerated three times with ethanol at room temperature. The mixture was then concentrated under reduced pressure, yielding an ethanol extract of 258.0 g. This extract was subsequently partitioned using hexane, chloroform, ethyl acetate, and water, producing a chloroform fraction (108.15 g), an ethyl acetate fraction (44.44 g), and a water fraction (105.18 g) ⁵⁵.

TABLE 7: ANTI-HIV-1 INTEGRASE ACTIVITY OF ETOH EXTRACT AND FRACTIONS FROM DIOSCOREA BULBIFER

The ethyl acetate and water fractions of Dioscorea bulbifera bulbils yielded seven compounds. Among these, allantoin, 2,4,3',5'-tetrahydroxybibenzyl, and 5,7,4'-trihydroxy-2-styrylchromone were identified for the first time in this plant. Myricetin exhibited the strongest activity with an IC50 value of 3.15 μM, followed by 2,4,6,7-tetrahydroxy-9,10-dihydrophenanthrene with an IC50 value of 14.20 μM, quercetin-3-O-β-D-glucopyranoside with an IC50 value of 19.39 μM, and quercetin-3-O-β-D-galactopyranoside with an IC₅₀ value of 21.80 μM. The potential interactions of the active compounds with the integrase (IN) active site were further explored. Compound 4 demonstrated the best binding affinity to IN, forming strong interactions with several amino acid residues, including Asp64, Thr66, His67, Glu92, Asp116, Gln148, Glu152, Asn155, and Lys159. These residues are involved in both the 3'-processing and strand transfer reactions of IN. Specifically, galloyl, catechol, and sugar moieties were identified as successful inhibitors of HIV-1 IN ⁵⁵.

Anti-Tumour Activity: Rhizomes of Dioscorea bulbifera were gathered in Qingyang, Anhui province, and authenticated by Prof. Shou-Jin Liu from Anhui College of Traditional Chinese Medicine in Anhui, China. A voucher specimen has been preserved in the herbarium of the Institute of Traditional Chinese Medicine at Shanghai University of Traditional Chinese Medicine ⁵⁶. The antitumor activities of the water extract (fraction A), ethanol extract (fraction B), ethyl acetate extract (fraction C), non-ethyl acetate extract (fraction D), and the compound diosbulbin B, isolated from Dioscorea bulbifera L., were investigated. Each mouse, except those in the normal group, was subcutaneously injected with approximately 10^6 S180 or H22 ascites tumor cells into the right armpit, following a reported experimental procedure. One day post-inoculation, the tumor-inoculated mice were randomly divided into several groups of ten. Mice treated with 5-fluorouracil (5-Fu) received intraperitoneal injections of the drug at a dose of 25 mg/kg every other day as a positive control. Both the normal (non-tumor-inoculated) and control (tumor-inoculated) groups were administered 0.5% CMC-Na solution (0.2 ml/10 g) orally each day. The treated groups were given specific concentrations of fraction A (275 mg/kg), fraction B (200 mg/kg), fraction C (40 mg/kg), and fraction D (160 mg/kg) via intragastric administration for 14 days, starting 24 hours after tumor inoculation. On the 15th day, blood samples from selected groups were collected from the retro-orbital plexus for hematological analysis. All animals were euthanized by cervical dislocation, and the tumors, spleens, and thymus glands were removed, weighed, and photographed. The tumor inhibition ratio (%) was calculated using the formula:

Tumor inhibition ratio (%) = [(C - T)/C] × 100,

Where, C represents the average tumor weight of the control group (treated with CMC-Na), and T is the average tumor weight of the treated group. The compound diosbulbin B, extracted from the rhizome of Dioscorea bulbifera, was found to inhibit the growth of transplanted S180 sarcoma in mice. Diosbulbin B significantly inhibited tumor growth in a dose-dependent manner, with tumor inhibition ratios of 45.76%, 65.91%, 77.61%, and 86.08% at doses of 2, 4, 8, and 16 mg/kg, respectively (P < 0.05) ⁵⁶.

Gastro Protective Activity: The hydroalcoholic extract of Dioscorea bulbifera tubers demonstrated gastroprotective activity at doses of 100, 200, and 400 mg/kg against indomethacin-induced gastric ulcers in rats ⁵⁷.

Neuro Pharmacological Activity: The hydroalcoholic extract of Dioscorea bulbifera tuber exhibited central nervous system depressant effects at doses of 100 and 300 mg/kg (administered orally). The treatments significantly reduced spontaneous motor activity and rectal temperature, and prolonged pentobarbitone-induced hypnosis in mice. However, the extract did not affect motor coordination as evidenced by the rota rod test, indicating a central rather than peripheral mechanism of action. Additionally, the extract showed anxiolytic activity in both the plus maze test and head-dip test ⁵⁸.

Anti-Diabetic Activity: Type II diabetes is characterized by an increase in blood glucose levels after meals. This postprandial glucose surge can be mitigated by inhibiting the activities of α-amylase and α-glucosidase, enzymes involved in carbohydrate digestion. Crude extracts of Dioscorea bulbifera have been shown to significantly inhibit α-amylase and trypsin activities (13.2 ± 2% and 4.3 ± 0.2%, respectively). Additionally, aqueous extracts of Dioscorea bulbifera tubers demonstrated antihyperglycemic effects in C57BL/6J mice and streptozotocin (STZ)-treated Wistar rats. When STZ-treated Wistar rats were administered 500 mg/kg doses of Dioscorea bulbifera extracts, significant antihyperglycemic effects were observed after six weeks of treatment. Various solvent extracts (ethyl acetate, methanol, petroleum ether, and 70% ethanol) of Dioscorea bulbifera were evaluated for their antidiabetic properties, with the ethyl acetate extract showing optimal results by inhibiting α-glucosidase by 99.6% and α-amylase by 73.39%. In a study by Ghosh and colleagues, diosgenin derived from Dioscorea bulbiferawas identified as a promising new drug candidate for type II diabetes. Ethyl acetate extracts of Dioscorea bulbifera containing diosgenin exhibited the highest inhibition rates of 82.64 ± 2.32% for α-glucosidase and 72.06 ± 0.51% for α-amylase ⁵⁹.

Anti-viral Activity: Concerning the pharmacological properties of Dioscorea bulbifera, only a limited number of studies have explored its antiviral activities. Various fractions of ethanolic extracts, including butanol, ethyl acetate, acetone, and ether, have been found to inhibit the coxsackie B1-VI virus ⁶¹. The ethyl acetate and acetone fractions were identified as the most effective in inhibiting viral activity compared to other fractions. Researchers also suggested that ethanol extracts at concentrations of 0.017-0.034 mg/ml from Dioscorea bulbifera could suppress RNA virus transcription and eliminate DNA viruses. Diosgenin, a compound derived from D. bulbifera, has been shown to inhibit the replication of the Hepatitis C virus at low doses. Additionally, hyperoside has been found to inhibit Hepatitis B virus activity both in laboratory settings and in living organisms, without being toxic to the host cells.⁶²Quercetin increases the inhibition of tumor necrosis factor in a dose-dependent manner against vesicular stomatitis virus, encephalomyocarditis virus, and herpes simplex virus type 1.

CONCLUSION: The pharmacognostic examination of Dioscorea bulbifera L. highlights its notable medicinal properties and reinforces its importance in traditional medicine. This research identifies a range of phytochemical components, including alkaloids, flavonoids, and saponins, that are responsible for the plant's therapeutic benefits. The existence of these bioactive substances indicates potential anti-inflammatory, antioxidant, and anticancer effects, positioning Dioscorea bulbifera as a strong candidate for future pharmaceutical exploration. Moreover, the morphological and anatomical characteristics revealed in this study offer crucial information for the accurate identification and quality assessment of the plant, ensuring that medicinal products derived from it are both effective and safe. The ethnobotanical knowledge associated with Dioscorea bulbifera further highlights its enduring significance across cultures, particularly in treating conditions such as respiratory issues, diabetes, and gastrointestinal problems. In summary, Dioscorea bulbifera is an invaluable asset in herbal medicine, warranting additional investigation to uncover its pharmacological properties and mechanisms. By merging traditional practices with contemporary scientific methods, we can fully harness its therapeutic potential. This plant not only embodies a rich legacy of medicinal use but also offers a pathway to the development of new therapies that align with the increasing interest in natural remedies and holistic health. Future research should aim to isolate specific compounds and conduct clinical trials to validate its medicinal claims and broaden its applications in modern healthcare.

ACKNOWLEDGEMENT: Nil

CONFLICT OF INTEREST: Nil

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    How to cite this article:

    Vishwakarma Y, Pandey V, Shukla R, Dhanorya D, Bairagi G and Gupta V: “Pharmacognostic analysis of Dioscorea bulbifera L.: a gateway to its medicinal benefits”. Int J Pharmacognosy 2024; 11(11): 584-94. doi link: http://dx.doi.org/10.13040/IJPSR.0975-8232.IJP.11(11).584-94.

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