A REVIEW ON: AEGLE MARMELOS

About Authors:
Kishan Singh *, Krishn Kumar Agrawal
Institute of Pharmaceutical Research GLA University,
Mathura-281403 (U.P.) India.
*kishan.singh575@gmail.com

ABSTRACT
Herbal drugs are traditionally used in various parts of the world to cure different diseases. The Ayurvedic and Siddha medical systems are very famous medical practices in Indian traditional medicines. Over the last few ,researcher have aimed at identifying and validation plant derived substance for the treatment of various disease .similarly it has been already proved that various parts of plants such as leaf, fruits seeds etc.provide heath and nutrition promoting compounds traditional used against various disease. Aegle marmelos have been used in ethno medicine to exploit its’ medicinal properties including astringent, antidiarrheal antidysentric, demulcent, antipyretic, and anti-inflammatory activities. The present review aims to complete medicinal values of Aegle marmelos generated through the research activity using modern scientific approaches and innovative scientific tools.

REFERENCE ID: PHARMATUTOR-ART-1484

INTRODUCTION
History
Bael or Bengal quince is a deciduous sacred tree, associated with Gods having useful medicinal properties, especially as a cooling agent. This tree is popular in ‘Shiva’ and ‘Vishnu’ temples and it can be grown in every house. Its leaves are trifoliate symbolizing the ‘Thrimurthies’- Brahma, Vishnu, Shiva, with spear shaped leaflets resembling “Thrisoolam” the weapon of Lord Shiva. Many legends, stories and myths are associated with this tree. The leaflets are given to devotees as ‘prasadam’ in Shiva temples and as ‘Tulasi’ in Vishnu temples.

Distribution
Bael tree is native to India and is found growing wild in Sub-Himalayan tracts from Jhelum eastwards to West Bengal, in central and south India.

Documented Species Distribution
Native range: India Exotic range: Bangladesh, Egypt, Malaysia, Myanmar, Pakistan, Sri Lanka, Thailand

Local Names
English (bael fruit, Indian bael, holy fruit, golden apple, elephant apple, Bengal quince, Indian quince, stone apple); Burmese (opesheet, ohshit); French (oranger du Malabar, cognassier du Bengale, bel indien); German (Belbaum, Schleimapfelbaum, Baelbaum); Gujarati (bili); Hindi (baelputri, bela, sirphal, siri-phal, kooralam); Indonesian (maja batuh,maja); Javanese (modjo); Khmer (bnau); Lao (Sino-Tibetan) (toum); Malay (bilak,bel, bila,maja pahit); Portuguese (marmelos); Thai (matum, mapin, tum); Vietnamese (trai mam, mbau nau)
Yield: The average yield is 300-400 fruits per tree. ¹
Family: Rutaceae

Botanical Description
Aegle marmelos is a slow-growing, medium sized tree, up to 12-15 m tall with short trunk, thick, soft, flaking bark, and spreading, sometimes spiny branches, the lower ones drooping. Young suckers bear many stiff, straight spines. A clear, gummy sap, resembling gum Arabic, exudes from wounded branches and hangs down in long strands, becoming gradually solid. It is sweet at first taste and then irritating to the throat. The deciduous, alternate leaves, borne singly or in 2's or 3's, are composed of 3 to 5 oval, pointed, shallowly toothed leaflets, 4-10 cm long, 2-5 cm wide, the terminal one with a long petiole. New foliage is glossy and pinkish-maroon. Mature leaves emit a disagreeable odor when bruised. Fragrant flowers , in clusters of 4 to 7 along the young branch lets, have 4 recurved, fleshy petals, green outside, yellowish inside, and 50 or more Greenish-yellow stamens. The fruits are round, pyriform, oval, or oblong, 5-20 cm in diameter, may have a thin, hard, woody shell or a more or less soft rind, gray-green until the fruit is fully ripe, when it turns yellowish. It is dotted with aromatic, minute oil glands. Inside, there is a hard central core and 8 to 20 faintly defined triangular segments, with thin, dark-orange walls, filled with aromatic, pale orange, pasty, sweet, resinous, more or less astringent, pulp. Embedded in the pulp are 10 to 15 seeds, flattened oblongs, About 1 cm long, bearing woolly hairs and each enclosed in a sac of adhesive, transparent mucilage that solidifies on drying ²

PHARMACOGNOSY OF PLANT

Macroscopic characters
A small to medium-sized aromatic tree, deciduous; stem and branches, light brown to green; strong auxiliary spines present on the branches; the average height of tree, 8.5 meters.³

Leaves
are alternate, pale green, trifoliate; terminal leaflet, 5.7 cm long, 2.8 cm broad, having a long petiole; the two lateral leaflets, almost sessile, 4.1 cm long, 2.2 cm wide, ovate to lanceolate having reticulate pinnate venation; petiole, 3.2 cm long.

Leaflets
Are ovate or ovate-lanceolate, margins crenate, apex acuminate, glabrous and densely minutely glandular-punctuate on both surfaces; lateral leaflets to 7 cm long and 4.2 cm wide, petiolules 0-3mm long.⁴

Flowers
Greenish white, sweetly scented, bisexual, actinomorpbic, ebracteate. hypogynous, stalked; stalk, 8 mm long; diameter of a fully open flower, 1.8 cal; flowers, borne in lateral panicles of about 10 flowers, arising from the leaf axil; calyx, gamosepalous, five-lobed, pubescent, light green, very small in comparison with petals; corolla polypetalous, with 5 petals, imbricate, leathery, pale yellow from above and green from beneath, length 4 mm; androecium, polyandrous, numerous, basifixed, 4 mm long, dehiscing longitudinally; gynoecium, light green, 7 mm long, having capitate stigma and terminal style.⁵

Stamens
Numerous; another elongate, apiculate; filaments free or fascicled, inserted round an inconspicuous disk. Ovary ovoid, cells 10-20; style terminal, short, deciduous; stigma capitate; ovules numerous, 2-seriate.

Fruits
yellowish green, with small dots on the outer surface, oblong to globose, 5.3 cm to 7 2 cm in diameter; weight, 77.2 g; volume, 73.7 ml; pulp, yellow and mucilaginous, the pulp of dried fruits retains its yellow, and also remains intact; rind woody, 4 to 5 mm thick.

Seeds
numerous, embedded in the pulp, oblong, compressed, white, having cotton-like hairs on their outer surface. seeds numerous, oblong, compressed, embedded in sacs covered with thick, orange coloured sweet pulp root bark is 3 to 5 cm thick covered, with creamy yellowish surface. It has a firm leathery texture, a sweet taste and fracture is fibrous. Stream bark is extremely gray and internally cream in colour. The outer surface is rough warty due to a number of lenticels, ridges and furrows. It is 4-8 mm thick, film in texture and occurs as flat or channeled pieces6. The fracture is tough and gritty in outer region and fibrous in the inner.5The taste is sweet and there is no characteristic odour5.

Chemical constituents
Various chemical constituents were found in bael like alkaloids, coumarins, steroids, polysaccharides, tannins, carotenoids etc.

Alkaloids
Agelin, aegelenine, marmeline, dictamine, fragrine, O-methylhalfordinine, O-isopentanylhalford iniol, N-4-methoxy styryl cinnamide.

Coumarins
Marmelosin, marmesin, imperatorin, marmin, alloimperatorin, methylether, xanthotoxol, scoparone, scopoletin, umbelliferone, psoralen and marmelide.

Polysaccharides
Galactose, arabinose, uronic acid and L-rhamnose was obtained on hydrolysis.

Tannin
Tannin was also present in leaves and fruit as skimmianine. Carotenoids were also reported, which import pale colour to fruit

Seed oil
Composed of palmitic, stearic, oleic, linoleic and linolenic acid. The fruit pulp contains 60.7 per cent moisture. The pulp contains 0.46 per cent acidity, 8.36 per cent total sugars, 6.21 per cent reducing sugars, 2.04 per cent non-reducing sugars and 0.21 per cent tannins. The pectin content is 2.52 per cent, which is quite high. The fruit pulp, however, is not a good source of vitamin C which is only 920 mg per 100 g of pulp4. This fruit is a very good source of protein which is 5.12 per cent of the edible portion. The total mineral content of the edible portion, as represented by ash, is 2.663 per cent. The percentage content of some of the minerals, viz. phosphorus, potassium, calcium, magnesium and iron is 0.137, 0.746, 0.188, 0.127 and 0.007 respectively.^{6, 7, 8}

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Plant parts used

Leaves
Leaves are applied to inflamed parts and are very efficacious in the form of poultice to unhealthy ulcers. Young leaves are eaten and said to cause sterility or even abortion. Juice of fresh leaves has a laxative action and also employed in asthmatic complaints, opthalmia and other eye affections. Decoction of leaves is used as a febrifuge and expectorant. Medicated oil prepared from leaves gives relief from recurrent cold and respiratory infections. The juice extracted from leaves is mixed with equal quantity of sesame oil and heated thoroughly, a few seeds of black pepper and half a teaspoonful of black cumin are added to the hot oil, and then it is removed from the fire and stored for use when necessary. A teaspoonful of this oil should be massaged onto the scalp before a head bath. Its regular use builds up resistance against cold and cough. Leaves are also use in Abscess, backache, abdominal disorders, vomiting, cut and wounds, dropsy, beriberi, weakness of heart, cholera, diarrhea, cardio tonic, blood sugar, injuries caused by animals, nervous disorders, hair tonic, acute bronchitis, child birth (George et al., 2003). Veterinary medicine for wound, killing worms, fodder for sheep, goat and cattle, stimulation of respiration contraction of denervosed nictitating membrane in anaesthetized cats (Gaur, 1999).

Root (bark)
Root bark is used in intermittent fever and as fish poison, as a remedy for palpitation of heat and melancholia. Juice of the bark with a little cumin in milk is valued as remedy for poverty of seminal fluid. The alcoholic extract of roots having hypoglycemic activity (Ohashi et al., 1995). It is also used in dog bite, gastric troubles, heart disorders, intermittent fevers, antiamoebic, hypoglycemic, rheumatism (Veerappan et al., 2000).

Flower
Distillation of flowers yielded a drug used as tonic for stomach and intestine, anti-dysenteric, antidiabetic, diaphorectic and as local anesthetics (Rahman and Ahmad, 1986). It is also used in epilepsy and as expectorant.

Fruit
Fruit is eaten during convalescence after diarrhea. It is valid for its mild astringency and as remedy for dysentery. The traditional healers of southern Chhattisgarh use dry powder of fruit with mustard oil for the treatment of burn cases. One part of powder and two part of mustard oil are mixed and is applied externally (Parmar and Kaushal, 1982). Fruits are also used in diarrhea, gastric troubles, constipation, laxative, tonic, digestive, stomachic, dysentery, brain and heart tonic, ulcer, antiviral, intestinal parasites, gonorrhea, epilepsy (Veerappan et al,2000), toys, edible, jam, preserve (Kaushik and dhiman, 1999).

Ripe fruit
The ripe fruit promotes digestion and is helpful in treating inflammation of rectum. The ripe fruit extract showed antiviral activity against ranikhet disease virus (Mazumdar, 1995). Pulp of ripe fruit is sweet, cooling, aromatic and nutritive when taken fresh. Fruit pulp marmalade is used as prevention during cholera epidemics, also given to prevent the growth of piles, useful in patients suffering from chronic dysenteric condition characterized by alternate diarrhea and constipation relieves flatulent colic from a condition of chronic gastrointestinal eatarrh. Fress juice is bitter and pungent fruit extract lower the blood sugar (Vyas et al., 1979).

Unripe fruit
Fine powder of unripe fruit showed significant effect on intestinal parasites and also effective against Entamoeba histolytica and Ascaris lumbricoides (Trivedi et al, 1978). Unripe fruit is used as an astringent in dysentery, stomachache in diarrhea, tonic, digeetive, demulcent, described as cardiacal, restorative, given in piles, Decoction of unripe fruit is astringent, useful in diarrhea and chronic dysentery.

Seed
Seed oil exhibits antibacterial activity against different strains of vibrios and inhibits the growth of Vibrio cholerae, Staphylococcus aureus and Escherichia coli (Banerji and Kumar, 1949). Essential oil exhibits antifungal activity against fungi Physalospora tucumanesis, Eeratocystis paradoxa, Selerotium ralfsii, Curvularia lunata, Helminthosporium sacchari, Fusarium monthforme and cephalosporium sacchari (Jain, 1977).

Chemical compounds isolated from plant
Leaf- Skimmianine, Aeglin, Rutin, Y-sit sterol, β-sitosterol, Flavone, Lupeol, Cineol, Citral, Glycoside, O-isopentenyl , Halfordiol, Marmeline, Citronellal, Cuminaldehyde phenylethyl cinnamamides, Eugenol, Marmesinin

Fruit-Marmelosin, Luvangetin, Aurapten, Psoralen, Marmelide, Tannin

Bark-Fagarine, Marmin.

Seed-Essential oil: D-limonene, A-D-phellandrene, Cineol, Citronellal, Citral, P-cyrnene, Cumin aldehyde.

Photochemical
The dry pulp of fruit contains chiefly mucilagepection like substance. The root, stem and leaves have been shown to contain tannins. Alkaloids, sterols, coumarin and aromatic components have been isolated. Aegelin, marmelosine, marmelin, o-methyl hayordinol, alloimperatorin methyl ester, o-isopentanyl hayordinol and linoleic acid have been identified.

Aegelin, formerly identified as sterol but clarified as a neutral alkaloid, rutacine, y-sitosterol, aegelemine and aegeline were identified from the leaves. Marmin, marmesin, umbelliferine, skimmianine were identified from the bark and roots. A major constituent of the fruit is the mucilage and marmelosin (0.5%) a coumarin, in addition to the minor constituents like reducing sugar essential oils, ascorbic acid and various minerals. A bitter, light-yellow oil contains 15.6% palmitic acid, 8.3% stearic acid, 28.7% linoleic and 7.6% linolenic acid. The seed residue contains 70% protein.Bael is reported to contain a number of coumarins, alkaloids, sterols and essential oils. Roots and fruits contain coumarins such as scoparone, scopoletin, umbelliferone, marmesin and skimmin. Fruits, in addition, contain xanthotoxol, imperatorin and alloimperatorin and alkaloids like aegeline and marmeline identified as N-2- hydroxy-2-[4 - (3’,3’-dimethyl allyloxy) phenyl] ethyl cinnamide. b- sitosterol and its glycoside are also present in the fruits. Roots and stem barks contain a coumarin - aegelinol. Roots also contain psoralen, xanthotoxin, 6,7- dimethoxy coumarin, tembamide, mermin and skimmianine. Leaves contain the alkaloids - O-(3,3- dimethyl allyl)-halfordinol, N-2-ethoxy-2 (4-methoxy phenyl) ethyl cinnamide, N-2-methoxy-2-(4-3’,3’-dimethyl allyloxy) phenyl] ethyl cinnamide, N- 2- [4-(3’,3’-dimethyl allyloxy) phenyl] ethyl cinnamide, N-2-hydroxy-2-[4-(3’,3’- dimethyl allyloxy) phenyl] ethyl cinnamide, N-4-methoxy steryl cinnamide and N-2-hydroxy-2-(4- hydroxy phenyl) ethyl cinnamide. Mermesinin, rutin and b-sitosterol - b-Dglucoside are also present in the leaves.^[9]A series of phenylethyl cinnamides, which included new compounds named anhydromarmeline (1), aegelinosides A and B were isolated from Aegle marmelos leaves as alfaglucosidase inhibitors. The structures of new compounds were characterized by spectroscopic data and chemical degradation of compounds isolated, anhydroaegeline (2) revealed the most potent inhibitory effect against alfaglucosidase with IC50 value of 35.8 lM. The present result also supports ethnopharmacological use of A. marmelos as a remedy for diabetes mellitus. ^[10]A rare alkaloid, shahidine (1), having an unstable oxazoline core has been isolated as a major constituent from the fresh leaves of Aegle marmelos. It is moisture-sensitive, and found to be the parent compound of aegeline and other amides; however, it is stable in dimethyl sulfoxide. Its structure was established by spectroscopic analysis. Biogenetically, oxazolines may be considered as the precursor of hydroxyl amides and oxazoles found in plants. Shahidine showed activity against a few Gram-positive bacteria.¹¹From dry leaves of Aegle marmelos, four new alkaloids, N- 2-[4-(3′, 3′-dimethylallyloxy)phenyl] ethyl cinnamide, N-2- hydroxy-2-[4-(3′,3′-dimethylallyloxy)phenyl] ethyl cinnamide, N-4-methoxystyryl cinnamide and N-2- hydroxy-2-(4-hydroxyphenyl) ethyl cinnamide were isolated and characterized. Also isolated were aegeline and a purple compound whose structure has not yet been established.¹²From the unripe fruits of Aegle marmelos, a new alkaloid named marmeline was isolated and identified as N-2-hydroxy-2-[4-(3′,3′- dimethylallyloxy)phenyl] ethyl cinnamide. Aegline, imperatorin, alloimperatorin and xanthotoxol were also present.^[13]The purified polysaccharide isolated from the cambium layer of a young bael (Aegle marmelos) tree contains galactose, arabinose, rhamnose, xylose, and glucose in the molar ratios of 10.0:9.8:1.4:1.9:1. Methylation analysis and Smith degradation studies established the linkages of the different monosaccharide residues. The anomeric configurations of the various sugar units were determined by oxidation of the acetylated polysaccharide with chromium (VI) trioxide. The oligosaccharides isolated from the polysaccharide by graded hydrolysis were characterized. The structural significance of these results is discussed. ^[14]The crude carbohydrate material isolated from bael (Aegle marmelos) seeds was resolved into four fractions. The homogeneous fraction contained 38.5% of carbohydrate and 60.6% of protein, and its carbohydrate moiety consisted of glucose, galactose, rhamnose, and arabinose in the molar ratios of 40:3:1:2. The linkages among various monosaccharide residues were established through methylation analysis and Smith-degradation studies. The anomeric configurations of the glycosyl groups and the structure at the glycosyl-amino acid junction were also determined. From the results of these experiments, a partial structure of the glycoprotein has been proposed.¹⁵Purified hemicellulose isolated from a young bael (Aegle marmelos) tree with 2.5m sodium hydroxide contained d-xylose and 4-O-methyl-dglucoronic acid in the molar ratio of 7.43:1; traces of glucose, galactose, rhamnose, and arabinose were also present. The linkages between the monosaccharide units were determined by methylation analysis of a hemicellulose fraction (II A) and carboxyl-reduced, hemicellulose II A, and the results were corroborated by those from periodate oxidation and Smith degradation. The anomeric configurations of the d-xylopyranosyl residues were determined by chromium (VI) trioxide oxidation of the acetylated, carboxyl-reduced hemicellulose, and the aldobiouronic acid obtained from graded hydrolysis was characterized. These experiments clearly revealed the structure of this hemicellulose.¹⁶The homogeneous, neutral polysaccharide isolated from the crude polysaccharide of the fruit pulp from bael (Aegle marmelos) contains arabinose, galactose, and glucose in the molar ratios of 2:3:14. The linkages among the different monosaccharide residues were established through methylation analysis and Smith-degradation studies of the polysaccharide. The anomeric configurations of the different glycosyl groups were determined by study of the chromium trioxide oxidation of the acetylated polysaccharide. Results of these experiments have been discussed in order to assess the structure of the neutral polysaccharide. ¹⁷A new 7- geranyloxycoumarin [7-(2,6-dihydroxy-7-methoxy-7- methyl-3-octaenyloxy) coumarin] named marmenol (1) has been isolated from the leaves of methanolic extract of Aegle marmelos belonging to the family Rutaceae. In addition to marmenol, several known compounds havealso been obtained for the first time from the same source. They include: praealtin D, trans-cinnamic acid, valencic acid, 4-methoxy benzoic acid, betulinic acid, N-pcis- and trans-coumaroyltyramine, montanine, and rutaretin. The structures of marmenol and known constituents were established with the help of NMR spectroscopy. However, structure of 1 was further confirmed via 2-D NMR experiments.¹⁸Antifungal constituents, 2-isopropenyl-4-methyl-1-oxacyclopenta[ b]anthracene-5,10-dione and (+)-4- (20- hydroxy-30-methylbut-30-enyloxy)-8H-[1,3]dioxolo[4,5- h]chromen-8-one in addition to known compounds imperatorin, b-sitosterol, plumbagin, 1-methyl-2-(30- methyl-but-20-enyloxy)-anthraquinone, b-sitosterol glucoside, stigmasterol, vanillin and salicin were isolated during phytochemical investigation on seeds of Aegle marmelos Correa.¹⁹

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Pharmacological properties:-

Nephroprotective activity
Present study was carried out to evaluate the Nephroprotective activity of an aqueous extract of Leaves of Aegle marmelos in Wistar rats. The aqueous extract of Aegle marmelos leaves was administered at three doses (250, 500, and 750 mg/kg, p.o.) to Wistar rats in Gentamicin induced nephrotoxicity model. The rats were pre-fed experimental diets for 8 days and then received GM (100 mg/kg body weight/day) treatment for 8 days while still on diet. Serum parameters, oxidative stress in rat kidney were analyzed. GM nephrotoxicity was recorded by increased serum creatinine and blood urea nitrogen level. GM increased MDA level whereas decreased catalase reduced glutathione level, while AEAM significantly reduced the elevated MDA levels and increased GSH and catalase concentration. GM increased serum creatinine, urea and blood urea nitrogen level, while AEAM reduced serum creatinine, urea and blood urea nitrogen level in gentamicin toxicity indicating a nephroprotective effect. The aqueous extract of A. marmelos leaves possesses the Nephroprotective activity.²⁰

Anti-ulcer activity
The present study was aimed to investigate anti-ulcer activity of methanolic and aqueous extract of Aegle marmelos seeds using indomethacin induced ulceration, stressed induced ulceration and pylorus ligation induced ulcerations. Ranitidine (50 mg/kg) was used as standard antiulcer agent. The ulcer index and percentage protection was estimated in all three models. Volume of gastric secretion, free acidity, total acidity and pH was estimated in pylorus ligation induced ulcer model. Methanolic extract showed significant (p<0.01) ulcer protective action at the doses of 200 and 400 mg/kg b.w. in all animal model. The aqueous extract was also found to possess significant (p<0.05) ulcer healing property at the same doses as of methanolic extract. A significant reduction in volume of gastric juice, free acidity and total acidity, along with increase in pH was observed in pylorus ligated rats. The antiulcer property of both the extracts was attributed due to the presence of quercetin like (Flavonoid) contents. ²¹

Hepatoprotective
In the present research studies, Bael leaves (Aegle marmelos, family of Rutaceae) which are also called as Bilva in ancient Sanskrit was used as herbal drug and its hepatoprotective effect in alcohol induced liver injury in albino rat was evaluated using essential biochemical parameters. The experiments were performed with four groups of animals.The experimental animals were administered with 30% ethyl alcohol for a period of 40 days and the fine crude plant leaves powder was fed to animals for next 21 days. The observed values of TBARS (Thiobarbituric acid reactive substances) in healthy, alcohol intoxicated and herbal drug treated animals were 123.35, 235.68 and 141.85 μg/g tissue respectively. The results were compared with the standard herbal drug silymarin (133.04 μg/g tissue). The experimental results indicate that, the Bael leaves have excellent hepatoprotective effect. A similar experimental Result was also observed in other biochemical parameters.^[22]

Physico-chemical analysis of ethnolic extracts from leaves of Aegle marmelos
The present study was carried out in physico-chemical analysis of ethnolic extracts from leaves of Aegle marmelos variants. It is commonly called Vilvam in Tamil. The leaves of Aegle marmelos variant-I and variant-II of the plant were collected and investigated for their phyto-chemical. The leaves of variants showed distinct variations. Anti microbial activity of ethanolic extracts both the variants of A.marmelos showed positive result against tested organism in a concentration dependent manner. ²³

Folk therapy
Aegle marmelos L. belongs to the family Rutaceae; an important medicinal plant being used in the folk therapy. It is the most useful medicinal plant of India. Its medicinal properties have been described in the ancient medical literature. All parts of the tree (stem, bark, root, leaves and fruit at all stages of maturity) have medicinal virtues and have been used as medicine for a long time. A number of chemical constituents have been isolated from various plant parts including alkaloids, coumarin and steroids. The leaves contain skimianinc, sterol and aegelin. In the present investigation antioxidant and hepatoprotective activity of the methanolic extract of A. marmelos leaves was examined on carbon tetrachloride (CCl4) intoxicated rats. MEAML hold considerable amount of phenolic (9.8367±0.0235 mg/kg) and flavonoid (8.248 ± 0.029 mg/kg) contents, which confirmed the antioxidant property of the leaves. The MEAML with different doses (50, 100, 200 mg/kg body weight) and standard silymarin (40 mg/kg body weight) were orally administered to CCl4 treated rats and the effect was studied on serum enzymes (AST), (ALT), alkaline phosphatase (ALP), and bilurubin, protein, albumin, thiobarbutiric acid (TBARS), reduced glutathione (GSH)]. In addition, in-vitro antioxidant activity of MEAML was also evaluated using (DPPH) radical. We noticed a significant hepatoprotective activity of MEAML on CCl4 intoxicated rats which support folkloric utilisation of A. marmelos, and it was further confirmed by the histological investigation. The findings of the present investigation revealed that the MEAML possess significant hepatoprotective activity by suppressing CCl4 induced cellular oxidative stress.²⁴

Antibacterial activity
Three medicinal plant Aegle marmelos, Lawsonia inermis, Albizzia libbeck were extracted by soxhlet apparatus using petroleum ether, ethanol, chloroform and aqueous as solvent. Among those extract, the petroleum ether was considered as effective one. The extracts were subjected to preliminary phytochemical screening and the three plants with four extracts were tested against three Gram positive bacteria (B.cereus, B.subtilis, S. aureus) and three Gram negative bacteria (E.coli, P.vulgaris, and P.aeruginosa) by disc diffusion method. Maximum inhibition (3.8cm) was recorded in Lawsonia inermis. It also showed inhibitory action against all the six pathogen tested. The zone of inhibition of the extracts was compared with the standard antibiotics Streptomycin and Spectinomycin. The study suggests that the plant is promising the development of phytomedicine for antimicrobial properties. ²⁵

Antidiabetic activity
Diabetes mellitus is associated with disturbances of learning and memory and cognitive functioning. Aegle marmelos Corr. from Rutaceae family, is widely used in Iranian folk medicine for the treatment of diabetes mellitus. It decreases blood glucose level by improving glucose tolerance and also has lipid-lowering and antioxidant properties. Considering the beneficial antidiabetic potential of A. marmelos, this study was conducted to evaluate the effect of chronic oral administration of A. marmelos as cognitive enhancer, on learning and spatial memory in diabetic rats using Morris water maze test. Male Wistar rats were randomly divided into normal-control, diabetic-control, and A. marmelos-treated diabetic groups (100, 250 and 500 mg/kg, p.o., 4 weeks). Diabetes was induced by a single dose i.p. injection of streptozotocin (45 mg/kg). In each group of animals, spatial learning and memory parameters were analyzed. A. marmelos showed dose dependent improvement in spatial learning and memory parameters. Swimming time (Escape Latency) in normal-control and A. marmelos-treated diabetic animals rats was significantly lower than diabetic-control, while swimming speed was significantly higher. The study demonstrated that A. marmelos has a significant protective effect against diabetes-induced spatial learning and memory deficits. This effect can be attributed to hypoglycemic, hypolipidemic and antioxidant activity of A. marmelos. ²⁶

Antigenotoxic activity
The present paper deals with the antigenotoxic activity of Aegle marmelos fruit extracts employing short term assays i.e. the SOS chromotest using Escherichia coli PQ37 and the Comet assay in peripheral human blood lymphocytes. Methanol extract and Acetone extract were quite effective in decreasing the SOS response induced by hydrogen peroxide and aflatoxin B1 in the SOS chromotest. Methanol extract inhibited the genotoxicity of H2O2 by 70.48% and that of AFB1 by 84.65%. The extracts showed significant decrease in the tail moment induced by hydrogen peroxide (9μM) in the Single Cell Gel Electrophoresis (SCGE) assay. The antigenotoxic activity exhibited by the extracts may be attributed the various polyphenolic constituents present in these extracts.²⁷

Studied the effects of skimmianine on the histamine release from rat mast cells
Skimmianine is a quinoline alkaloid isolated from the roots of Aegle marmelos Correa. In the study, we studied the effects of skimmianine on the histamine release from rat mast cells. The study was performed by using two cell lines, rat basophilic leukemia (RBL-2H3) cell line, and rat peritoneal mast cells (RPMCs). DNP24-BSA, thapsigargin, ionomycin, compound 48/80 and PMA were used as inducers for histamine release from rat mast cell. Skimmianine markedly inhibited the histamine release from RBL-2H3 cells induced by DNP24-BSA, thapsigargin and ionomycin. The effec suggested is related to Ca2+ signaling since skimmianine showed strong effects when the histamine release induced by Ca2+ signal stimulants (thapsigargin and ionomycin). It is supported that skimmianine altered the influx of 45Ca2+ into the cells. In RPMCs experiment, skimmianine also suppressed the histamine release induced by Ca2+ stimulants, and phorbol myristate acetate (PMA). However, skimmianine had no effect on the histamine release induced by compound 48/80. Based on the results, the inhibitory effects of skimmianine on the histamine release from mast cells might involve some mechanisms related to intracellular Ca2+ signaling events and protein kinase C signaling possessing a main role in granule exocytotic processes

Ulcer protective
The effect of 50% ethanolic extract of Aegle marmelos (AM) root was assayed in different acute and chronic gastric ulcer model in rats. AM, 200-400mg/kg administered orally, twice daily for 5 days showed dose dependent ulcer protective effect in pylorus ligation (19.3-69.5% protection, p<0.05 to p<0.001), Aspirin + PL (24.4- 68.0 % protection, p<0.001), ethanol (26.0 – 72.0 % protection, p< 0.001), cold restrain stress (31.6 -65.4% , p< 0.01 to p< 0.001) and acetic acid (16.8- 90.4% protection, p<0.001). The effect of extract on gastric content volume, ph, acidity, using PL models was also evaluated. Regarding the models of gastric secretion, a reduction on volume of gastric juice and acidity was observed as well as increase in gastric pH. The extract was screened for possible antioxidant activities by free radical scavenging activity (DPPH), hydroxyl radical, hydrogen peroxide scavenging activity (HPSA). These results clearly indicate that aerial parts of Aegle marmelos were effective in scavenging free radicals and has the potential to be a powerful antioxidant. However, AM reduced the ulcer index with significant decreasein lipid peroxidation (27.9 – 55.4% protection, p< 0.001), superoxide dismutase (23.0- 41.5% protection p< 0.01 and 0.001) and increased in catalase (35.9 – 79.8% protection p< 0.01 and p< 0.001) activity respectively. Preliminary phytochemical screening of AM gave the positive tests for flavonoids, steroids, alkaloids,terpenoids, saponins and tannins etc. The purpose of present study was to study the safety and efficacy of this extract against peptic ulcer diseases and provideprotection against free radical to gain the balance. ²⁸

Antimicrobial activity
Aegle marmelos is a medicinal herb belongs to the family Rutaceae. The different parts of plants like leaves and flowers are extracted by using the solvent methanol. The methanol extracts were screened for the antimicrobial activity. They showed greater inhibitory effect against both gram positive and gram negative organisms. The organisms used were such as Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis, Salmonella typhi, Staphylococcus aureus. Based on the present investigation results it is concluded that the methanolic extracts of Aegle marmelos has great potential as antimicrobial agent against different microorganisms and they can be used in the treatment of infectious diseases caused by the resistant microorganisms. ²⁹

Antibacterial activity:-
The principal objective of the present research work was to determine the antibacterial potential of Aegle marmelos against seven standard pathogenic bacterial strains. To evaluate antibacterial activity the agar-well diffusionassay was used. All the three extracts showed the highest and significant antibacterial activity against both Gramnegativeand Gram-positive bacteria. It is the strain Bacillus subtilis that is almost resistant to the three extracts of Aegle marmelos.³⁰

Antioxidant Activity
Antioxidants are the compounds with free radicals scavenging activity and capable of protecting the cells from free radical mediate oxidative stress. The antioxidant compounds can be derived from natural sources such as plants. Antioxidant activity of these plants is due to the presence of flavones, isoflavones, flavonoids, anthocyanin, coumarin lignans, catechins and isocatechins. A. marmelos is extensively reported to possess antioxidant activity against a variety of free radicals. Antioxidant activity of the fruit of A. marmelos was reported. Antioxidant activity and free radical scavenging activity of the ripe and unripe fruit of Aegle marmelos was compared. Results indicate that the enzymatic antioxidants increased in ripe fruit when compared to unripe fruit extract (except glutathione peroxidase). The percentage of free radical inhibition was also high in unripe fruit than that of the ripe fruit ³¹. Methanol and aqueous extract of A. marmelos fruit pulp was screened for antioxidant activity by DPPH radical scavenging method, reducing power assay, nitric oxide scavenging assay, superoxide radical scavenging assay, ABTS radical scavenging assay and H2O2 radical scavenging assay. Both aqueous and alcoholic extract exhibited good antioxidant activity.³²The antioxidant activity of the fruit of A. marmelos was reported. The aqueous extract of A. marmelos fruit was screened for antioxidant activity by the DPPH radical scavenging. The extract showed efficient antioxidant activity.³³

Ant diarrheal Activity
Antidiarrheal activity is one of the major medicinal properties of A. marmelos and traditionally it is extensively used to control chronic diarrhea and dysentery. Recently, several in vitro and in vivo studies have been conducted to confirm the antidiarrheal property of A. marmelos. The in vitro antidiarrheal activity of dried fruit pulps of A. marmelos was reported. Antidiarrheal activity was performed by MIC method against the causative organisms of diarrhea. The ethanolic extract showed good activity against Shigella boydii, S. sonnei and S. flexneri, moderate against S. dysenteriae³⁴. Crude aqueous extract of unripe fruits of A. marmelos was screened for causative agents ofdiarrhea. The extract was analyzed for antibacterial activity, antigiardial activity and antirotaviralactivity. The extract exhibited inhibitory activity against Giardia and rotavirus whereas viabilityof none of the six bacterial strains tested was affected. ³⁵

Antiproliferative activity
The different solvent fractions of ethanol extract of the stem barks of A. Marmelos were Reported to possess antiproliferative effects against human tumor cell lines. The results showed The inhibition of in vitro proliferation of human tumor cell lines, including the leukemic K562, Tlymphoid Jurkat, Blymphoid Raji, erythroleukemic HEL, melanoma Colo38, and breast cancer MCF7 and MDAMB-231 cell lines. ³⁶

Cytoprotective Effect
The cytoprotective effect of the leaves of Aegle marmelos was reported in Cyprinus carpio (freshwater fish) exposed to heavy metals. C. carpio was exposed to heavy metals followed by the treatment with the dried powder of Aegle marmelos leaves. Treatment resulted in cytoprotective effect by stabilization of plasma membrane and modulation of antioxidant enzyme System. ³⁷

Antifertility Effect
The antifertility effect of the aqueous extracts of leaves of Aegle marmelos was reported in male Albino rats. The rats were administered with aqueous extracts (250 mg/kg body weight) of leaves of Aegle marmelos for 45 days. Treatment resulted in reduction in the weights of testis, epididymes and seminal vesicle. The extract also resulted in reduction of testicular sperm count, epididymal sperm count and motility and abnormal sperm count. ³⁸

Insect controlling properties
Essential oil from the leaves of A. marmelos was reported for showing insecticidal activity against four stored grain insect pests included Callosobruchus chinensis (L.), Rhyzopertha dominica (F.), Sitophilus oryzae (L.) and Tribolium castaneum. In the study grains were infected with test insects, and were fumigated with essential oil of A. marmelos . The oil treatment significantly reduced the grain damage as well as weight loss in fumigated grains samples infested with all insects except T. castaneum. The essential oil at different doses significantly reduced oviposition and adult emergence of C. chinensis in treated cowpea seeds^[39].Essential oil from the leaves of A. marmelos was reported for insect repellent activity against Sitophilous oryzae and Tribolium castaneum. However A. marmelos essential oils didn’t showed 100% repellent activity against the test insects. ⁴⁰

Analgesic activity
Leaves of A. marmelos were reported to possess analgesic activity. Methanol extract of leaves of A. marmelos was screened for analgesic activity by Acetic acid-induced writhing test in Swiss mice. The results indicated that methanol extract significantly reduced the writhing induced by acetic acid. In tail flick test methanol extract (200 and 300 mg/kg body weight) showed significant analgesic activity in the. ⁴¹

Antiarthritis activity
Leaves of A. marmelos were reported to possess antiarthritis activity against collagen induced arthritis in Wistar rats. Methanol extract treatment of rats showed the reduction of paw swelling and arthritic index. Radiological and histopathological changes were also significantly reduced in methanol extract treated rats.⁴²

Anti-inflammatory activity
Unripe fruit pulp of A. marmelos was reported to possess anti-inflammatory activity. Inflammation was induced by injecting 0.1 ml of 1% λ carrageenan into the sub planer side of left Hind paw of Sprague Dawley rats. Extract treatment of the inflammated rats significantly reduced the λ carrageenan induced inflammation.⁴³

Medicinal properties
The roots are useful for treating diarrhoea, dysentery, and dyspepsia¹, ⁴⁴. The leaf is used for opthalmia, diabetes, and asthmatic complaints. Unripe fruit is useful for treating diarrhoea, dysentery and stomachalgia. The aqueous extracts of the stem and root bark are used to treat malaria, fever, jaundice, and skin diseases such as ulcers, urticaria, and eczema⁴⁴, ⁴⁵.In pharmacological trials, both the fruit and root showed antiamoebic and hypoglycaemic activities⁴⁴, ⁴⁶, ³⁴. Aqueous leaf extract and methanolic extract of the root bark of A. marmelos showed preventive effects on myocardial diseases¹, ⁴⁵, ^{and 46}. The leaves are used to cure sinusitis, dyspepsia and anorexia6. This fruit was used to cure tuberculosis, loss of appetite, emaciation etc. There are several such pharmacopoeias in Siddha medicine. Aegle marmelos (Linn) family Rutaceae was highly reputed ayurvedic medicinal tree1. Various phytochemical and biological evaluations have been reported as anti-diabetic⁴⁷, ⁴⁸, antioxidant, antithyroid36. It is astringent, cooling, carminative, laxative, restorative and stomachic and is used in dysentery, diarrhoea, flatulence, fever, vomiting and colic. The leaves are astringent, laxative, febrifuge and expectorant and are useful in ophthalmia, deafness, inflammations, diabetes and asthmatic complaints. The tender fruit is bitter, astringent, antilaxative, digestive and promotes digestion and strength, overcomes vata, colics and diarrhoea. The ripe fruits are astringent, sweet, aromatic, cooling, febrifuge, laxative and tonic and are good for the heart and brain. Antidiabetic property, antidiarrhoeal activity, antiulcer activity of seeds, antifungal activity of leaves and antitumour and antimutagenic activity of this plant are clinically evaluated, ⁵⁰,⁵¹

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Conclusion
It is quite evident from this review that Aegle marmelos contains a number of phytoconstituents which reveals its uses for various therapeutic purposes. Looking upon wide prospects and potential of bael for various purposes, this will help in financial upliftment of the poor and landless farmers besides providing base for the Research and Development. The Plant or its individual parts can be used for the treatment of various disorders in human being such as, diabetes, liver toxicity, fungal infection, microbial infection, inflammation, pyrexia and to relieve pain. Still, so much work is required with the Aegle marmelos to investigate the mechanism of actions with other therapeutic activities.

ACKNOWLEDGEMENTS:
Authors are thankful to Mr. Jeetendra Kumar Gupta, Assistant Professor, Department of Pharmacology,GLA IPR University Mathura , Bhupesh Chander Semwal, Assistant Professor, Department of Pharmacology Mathura for their valuable support.

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