Evaluation of pharmacognostical, phytochemical and ethnobotanical properties of Euphorbia neriifolia

INTRODUCTION

Euphorbia neriifolia is a deciduous, euphorbiaceous medicinal plant (Sharma and Janmeda, 2013; Pokharenet. al, 2011; Toume et. al, 2012; Sing et. al, 2003). As a south Asian indigenous plant, beside Bangladesh, Sri Lanka and India, it is also found in Myanmar, Thailand and Malaysia (Ahmed et. al, 2011; Samaresh et. al, 2013; Mansuriet. al, 2013; Vimala and Shoba, 2014). Although commonly known as “Milk hedge”, it has several vernacular names as sehund, thohar, mansasij, patashij in India and Bangladesh (Bose GC, 1920; GRIN, 2014; Panda H, 1999; Katewa et. al, 2008; Pracheta et. al, 2011; Khare CP, 2007; Bigoniya and Rana, 2009). It is a succulent, fleshy, spiny, erect plant, about 3-6 meters long, having unarticulated stems and branches (Kumar et. al, 2012). Its leaves are apex rounded, base attenuated, oval shaped and fruits are three chambered whereas it’s male and female both flowers are found in same bunch (Sandeep et. al, 2013). In spite of its diverse curative role, it is still underutilized. Therefore, this effort has taken to promote its phytotherapeutical roles by highlighting its folk use and pharmacological as well as photochemical evaluation.

ETHNOBOTANICAL APPLICATION

As a significant medicinal plant, the traditional use of E. Neriifolia for curing many diseases has a long history (Rahmatullah et. al, 2012). Its juice familiar as milky latex has diverse ethno medical use. A succus compounded of equal parts of its juice is said to be used for relief in asthma. Equal parts of its juice with other compounds as formulation administered as a dose of 10 to 20 minims three times a day in asthma. Juice heated with or without the gum of E. resinifera, dropped into the ear to afford relief in earache in Malaysia. In Sarawak, instead of squeezing the juice out of the leaves they simply inserted rolled leaves into the ear canal and blew air through it. Milk-juice exuded from injured fleshy cylindrical stems is used by traditional physicians as medicine in relieving earache. Cloves, long-peppers, chebulic myrobalans and trivrit root are soaked in juice for some months after drying used in syphilis, dropsy, general anasarca, leprosy. It also used as drastic purgative in the enlargement of liver and spleen. Juice mixed with ghee is used in syphilis, visceral obstruction and liver enlargements. In ulcers and scabies, juice is used with fresh butter. Turmeric powder mixed with the juice is used in haemorrhoids and piles (Samaresh et. al, 2013; Panda H, 1999; Khare CP, 2007). The latex is also used to remove warts by direct application over the lesion. The juice come from the heated leaves is a remedy for otalgia and otitis. The tribal population of Chhattisgarh region in India used milky latex as aphrodisiac. Latex is also prescribed for skin diseases, urinary disorders and diabetes approved by sushrutasamhita. It is also used as medicine in rube-facient, chlorosis, tympanitis and cutaneous eruptions (Ahmed et. al, 2011). Natives of Chhattisgarh use it in castor oil with salt for deep cracks of legs. Milky latex is used commonly like aloe gel in case of burn (Bigoniya and Rana, 2009). Formulation prepared from root-bark, boiled in rice-water, is given in dropsy which also mixed with black-pepper to apply in scorpion stings and snake-bites. Pulp of its stem mixed with fresh ginger is used to prevent hydrophobia. A small dose of its roasted stem with honey is helpful to expectoration of phlegm (Panda H, 1999). Root and stem both are used as antiseptic (Ahmed et. al, 2011). The usages of its leaves are traditionally fruitful in the management of piles, aphrodisiac, diuretic, cough, cold, bronchitis (Ahmed et. al, 2011; Bigoniya and Rana, 2009). Formulated ‘’Kaajal’’ from the leaf of EN showed antimicrobial properties against eye pathogens for the present of carbon nanoparticles (Mandal et. al, 2013). The prominent traditional applications of different parts of it are summarized in Table 1.

Table 1: The Ethnomedicinal uses of Euphorbia neriifolia

Plant’s part	Used in/   Used as	Mode of application	Reference
Juice	Drastic purgative	Cloves, long-peppers, chebulicmyrobalans and trivrit root are soaked in juice for some months; then, dried and applied by heated with salt	Panda H, 1999;Khare CP, 2007
	Syphilis	Juice mixed with ghee and other compounds as formulation is given in syphilis.	Samaresh et. al, 2013;Panda H, 1999;Bigoniya andRana, 2009
	Cutaneous eruptions	NS	Ahmed et. al, 2011
	Asthma	Madar flowers, agadha root and gokaran root are steeped in its juice powdered form is given with honey and chebulicmyrobalans.	Panda H, 1999;Khare CP, 2007
	Rube-facient	NS	Ahmed et. al, 2011;Khare CP, 2007
	Whooping cough	Heated with salt	Panda H, 1999
	Earache	Leaves heated with or without gum of E.resinifera in earache in Malaysia. In Sarawak, rolling leaves insert it in the ear canal.	Ahmed et. al, 2011;Panda H, 1999
	Chlorosis	NS	Ahmed et. al, 2011
	Visceral obstruction	Juice mixed with ghee	Panda H, 1999
	Liver enlargements	Juice mixed with ghee	Panda H, 1999
	Warts remover	Directly	Ahmed et. al, 2011; Panda H, 1999
	Dropsy, Leprosy	Juice is dried with other plants as formulation and administered with salt	Samaresh et. al, 2013;Panda H, 1999; Bigoniya andRana, 2009
	Dyspepsia	Heated with salt	Panda H, 1999
	Ophthalmia	Mixed with soot of ghee-lamp
	Ulcers	Juice is largely used with fresh butter
	Scabies	Juice is largely used with fresh butter
	Glandular swelling	It prevents suppuration
	Rheumatic limbs	Mixed with Margosa oil
	Tympanitis	NS	Ahmed et. al, 2011
	Piles	Turmeric powder mixed with the juice	Panda H, 1999
	Diabetes	NS	Ahmed et. al, 2011
	Haemorrhoids	Turmeric powder mixed with the juice	Panda H, 1999
	Jaundice	Heated with salt	Panda H, 1999
	Otalgia, Otitis	The juice from the heated leaves is used as a remedy	Ahmed et. al, 2011
	Eye infection	NS	Mandal et. al, 2013
	Cracked legs	Natives of Chhattisgarh use it in castor oil with salt	Bigoniya andRana, 2009
	Aphrodisiac	NS	Ahmed et. al, 2011
	Burns	Used commonly like aloe gel	Bigoniya andRana, 2009
	Skin diseases	NS	Ahmed et. al, 2011
	Urinary disorders	NS	Ahmed et. al, 2011
Root	Dropsy	Root-bark boiled in rice-water	Panda H, 1999
	Antispasmodic	NS	Ahmed et. al, 2011
	Scorpion stings and snake-bites	Root mixed with black-pepper is employed in both superficially and orally	Panda H, 1999
Root	Antiseptic	NS	Ahmed et. al, 2011
Stem	Expectoration of phlegm	Stem is roasted in ashes and juice with honey; then, given with borax in small doses	Panda H, 1999
	Hydrophobia	Pulp of the stem is mixed with fresh ginger
	Passing of semen with urine	2g bark  macerated with 4 leaves of Piper betleL.and taken during evening or at night before falling asleep	Rahmatullah et. al, 2012
Leaf	Piles, fistula	NS	Ahmed et. al, 2011;Bigoniya andRana, 2009
	Aphrodisiac	NS	Ahmed et. al, 2011
	Diuretic	NS	Ahmed et. al, 2011
	Cough and Cold	NS	Ahmed et. al, 2011

 

Table 2: Nutrient & phytochemical investigation of Euphorbia neriifolia

Plant’s part	Phytochemical/   Nutrient	Identification procedure of used extract	Reference
Leaf	Proteins	Millon’stest (PE, C, EA, Aq),   Biuret test (EA, HE, PE, B, C, E), Xanthoproteinic test (HE, B, EA, E,Aq)	Yadav et. al, 2011;   Sharma and Pracheta et. al, 2013; Pracheta et. al, 2011
	Carbohydrate	Fehling’stest(HE, C, E)
	Fixed oil	Stain test (HE, PE, B, C, EA, E, Aq)
	Fat	Soap test(HE, E, PE, EA)
	Alkaloids	Dragendroff’s test(PE, B, EA, HE, C, Aq, E)
		Mayer’s test(HE, PE, B, C,E)
		Wagner’s test(HE, PE, C, EA, E)
	Anthraquinones	Borntrager’s test(HE, E, C, EA, Aq)
	Cardiac Glycosides	Killer Killani test (HE, E, PE, EA, B, C, Aq)
		Baljet test (HE, E, PE, EA, C, Aq)
	Glycoside	Legal’s test(B, C, E, Aq, HE, E)
	Flavonoids	Shinoda test ( HE, E, C, EA, Aq),   Ammonia test(E, B,HE, C, EA, Aq), Lead acetate test (HE,EA, E, Aq,C), Alkaline reagent test(E,HE,EA, Aq)
		Ammonia test (C, E, EA, Bu)	Pokharen et. al, 2011
	Phenolic compounds	Ferric chloride test (E, HE, ,EA, Aq)	Pokharen et. al, 2011;   Sharma and Pracheta et. al, 2013; Pracheta et. al, 2011
	Pholobatanins	Hydrochloride test(E, HE, Aq and C, E, EA)	Pokharen et. al, 2011;   Sharma and Pracheta et. al, 2013; Pracheta et. al, 2011
	Polyphenol	Folin-ciocalteau test(HE, EA, E, Aq, PE)	Sharma and Pracheta et. al, 2013
	Saponins	Frothing test(B,PE, C), Olive oil test(B, PE,HE, C, EA, E)	Sharma and Pracheta et. al, 2013;   Pracheta et. al, 2011
	Steroids	Liebermann-Burchard’s test(HE,C, E,PE, B)	Sharma and Pracheta et. al, 2013;   Pracheta et. al, 2011
	Tannins	Ferric chloride test(C, HE, E, EA, Aq)	Pokharen et. al, 2011;   Sharma and Pracheta et. al, 2013; Pracheta et. al, 2011
	Terpenoids	Salowski test(HE, E, PE, B, C EA)	Toumeet. al, 2012;   Sharma and Pracheta et. al, 2013; Zhao et. al, 2014
	Gums,Mucilage	RutheniumRed test(HE,E, Aq, B, EA,C,PE)	VimalaandShoba, 2014;   Sharma and Pracheta et. al, 2013
	Cardenolides	NS(C, EA, Bu)	Pokharen et. al, 2011
	Sapogenin	Column Chromatography (E)	Bigoniya et. al, 2010
Root	Terpenoids	NS(E)	Ng AS, 1989
Flower	Alkaloids	Wagners test (M, E),   Dragendorff’s test (M, E)	Leela et. al, 2013
	Flavonoids	Pews test (M, E),   Shinoda test (M, E), NaOH Test (M, E)
	Glycosides	Keller-Killani test (PE, C, M, E, AQ), Glycosides test (C, M, E),   Conc. H2SO4(PE, C, M, E, AQ), Molishs test (PE, C, M, E, AQ)
	Phenol	Ellagic test (M, E,PE, C, AQ),   Phenol test (M,E,PE,C, AQ)
	Lignin	Lignin test (C, M, E),   Labat test (C, M, E)
	Saponins	Foam test (M, E, AQ),   Haemolysis test (M, E)
	Sterols	Libermann-Burchard test (PE, C, M, E, AQ), Salkowski test (C, M, E, PE)
	Tannins	Gelatin test(M, E, PE, C, AQ),   Lead Acetate test(M, E, AQ, PE, C)
Stem	Alkaloids	NS(M)
	Flavonoids	NS(M)
	Glycosides	NS(M)	Samaresh et. al, 2013
	Saponins	NS(M)
	Terpenoids	NS(M)
Latex	Terpenoids   (Neriifolin S)	Ammonium sulfate precipitation model, Anion exchange Chromatography,   Size-exclusion Chromatography	Yadav et. al, 2011
	Terpenoids	H- NMR,   C-NMR, IR and mass spectra	Ilyas et. al, 1998
	Lectin	Affinity chromatography	Seshagirirao and Prasad, 1995
(HE: Hydro-ethanolic extract, PE: Petroleum ether extract; B: Benzene extract; C: Chloroform extract; EA:Ethyl-acetate extract; E: Ethanol extract, Aq: Aqueous extract; M: Methanol extract; Bu: Butanol extract)

 

Table 3: Pharmacological activities of various extracts of Euphorbia neriifolia 

Activity	Plant part	Extract	Experimental Details	Reference
Antibacterial	Stem	Methanol	400 mg/ml showed highest activity against Pseudomonas aeruginosa. MIC showed more efficacies against Staphylococcus aureus.	Samaresh et. al, 2013
	Leaf	Hydro   alcohol	50µg/ml inhibited the growth of P. vulgarius and K. pneumoniae.	Pokharen et. al, 2011
	Latex milk	Methanol	Inhibition zones were found against Salmonella typhi and Klebsiella pneumoni, respectively 8 mm by agar diffusion method at 60 μl concentration. Disc diffusion method provided 7 mm inhibition zone against Pseudomonas aeruginosa at 50 μl. MIC showed more efficacious against Salmonella typhi, Klebsiella pneumonia and Pseudomonas aeruginosa at higher concentration.	Sumathi et. al, 2012
Antifungal	Stem	Methanol	Extract at 400 mg/ml showed maximum inhibition area against Aspergillus nigur, Candida albicans as 12 and 14 mm, respectively in disc diffusion method.	Samaresh et. al, 2013
	Latex milk	Methanol	Spore germination assay (SGA) provided the reduced percentage of spore germination against Aspergillus flavus, Aspergillus fumigates and Mucor.	Sumathi et. al, 2012
Antiviral	Leaf	Ethanol	Isolated 3-β-friedelanol exhibited potent activity in anti-hcov test.	Chang et. al, 2012
	NS	NS	Eurifoloids E and F exhibited significant anti-HIV activities, with EC50 values of 3.58±0.31 and 7.40±0.94 μM , respectively.	Zhao et. al, 2014
Anti-inflammatory	Leaf	Hydro   alcohol	Indomethacin and extract (400 mg/kg) showed 63.27 and 55.12% activity in carrageenan-induced paw edema rat model.	Gaur et. al, 2009
	Leaf	Hydro-alcohol	The same rat model showed inhibition of edema volume 69.47 and 75.78%, respectively by extract at 400 mg/kg dose and aspirin. Same dose of extract and diclofenac showed 60.61 and 66.14% inhibition of granulomatous tissue mass development recognized by cotton pellet-induced granuloma model.	Bigoniya and Rana, 2010
Immuno-modulatory	Leaf	Hydro-alcohol	Betamethasone-induced immunosuppressive rat model displayed mortality 16.6% (48 hours) by extract (400 mg/kg). In carbon clearance method, phagocytic index of extract (400 mg/kg) was 0.0286. Treatment of extract at 400 mg/kg showed primary titre as 14.7±0.26, and secondary titre as 20.7±0.28 in haem-agglutination antibody titre method. Footpad swelling method (48 hours) of extract (400 mg/kg) determined delayed type hypersensitivity as 5.7±0.01.	Gaur et. al,   2009
Antioxidant and radical scavenging	Leaf	Ethanol	TBA method showed extracts and control decreased liver lipid peroxidase as 7.78±0.69 and 29.94±1.22 nmol/mg, respectively. It also increased SOD as 26.06±1.12, 47.58±2.24 unit/mg in liver and kidney.	Bigoniya and Rana, 2009
	Leaf	Hydro-alcohol	Ferric ions reduction ability was 149.2±0.05μmol detected by FRAP method. At 1mg/ml of extract, inhibition of superoxide generation was 50.06 in superoxide radical scavenging assay and scavenging activity was 76.15% in DPPH assay. The percentage of metal chelating capacity of extract and standard was 73.24 and 85.37%, respectively. The percentages of H2O2 scavenging activity of extract, ascorbic acid and BHT were found to be 69.01, 12.7 and 44.7%, respectively.	Pracheta et. al, 2011
	Leaf	NS	Total sapogenin fraction of extract and α-tocopherol showed 190.36±2.86 and 447.7±6.21 ASE/mg reducing power activities, respectively by reducing power assay, whereas IC50 values respectively were 440.6±42.3 and 38.6±0.7 µg/ml in DPPH assay, 806.8±57.9 and 34.2±0.4 µg/ml in hydroxyl radical scavenging assay and 471.3±38.2 and 59.6±1.0 µg/ml in superoxide radical scavenging assay.	Bigoniya and Rana, 2009
	Leaf	Ethanol	TBA method showed SOD increase as 4.12±0.76 unit/mg at 175 mg/kg. Lipid peroxidase reduction ability was 14.70±1.22 nmol/g.	Bigoniya et. al, 2010
Antidiabetic	Leaf	Ethanol	Extract at 400 mg/kg with control produced 99.6±2.540 mg/dl fall of glucose at 60 min where control produced 110.2±3.01 mg/dl in high fat diet-streptozotocin induced type-2 diabetic model.	Mansuri et. al, 2013
	Leaf	Ethanol	Glucose oxidase model provided 43.23±3.58 mg/dl effect at 400 mg/kg where control group showed 112.63±4.68 mg/dl.	Bigoniya and Rana, 2009
Thrombolytic	Latex	NS	A lectin from latex showed thrombolytic activity.	Seshagirirao and Prasad, 1995
Hematological	Leaf	Ethanol   Petroleum ether, Acetone	Extract at 400 mg/kg increased Hb, neutrophil and lymphocyte as 14.50±0.74 gm/dl, 40.02±1.72% and 58.86±1.50% where control showed 12.20±0.97 gm/dl, 53.14±1.51% and 44.17±1.59%, respectively in 45 days.	Bigoniya and Rana, 2009
Anti-cancer	Leaf	Hydro-ethanol	DENA-induced renal carcinogenesis in mice model showed significant activity by restoring liver enzyme.	Sharma and Pracheta, 2013; Sharma et. al,2011; Pracheta et.al, 2011
		Ethanol	Radiation-induced chromosomal aberrations in cultured human lymphocytes model showed IC50 of total sapogenin inhibited growth of mouse melanoma cells by 50% was 173.78µg/ml compared to120 ng/ml for vincristine.	Bigoniya and Rana, 2009
	NS	Ethyl acetate	Isolated compound showed potent activity on Panc-1, 81T, BE3 and K562 cell line by MTT assay.	Lin et. al, 2013
Wound healing	Leaf	Methanol	Excision wound rat model showed the efficacy as 100.00±1.93% and 85.66±2.78%, respectively in case of extract at 500 mg/kg and control at 18th day. Incision breaking strength was found 700.31±3.67 by same dose extract in incision wound rat model.	Pattanaik et. al, 2014
	Leaf	Ethanol	The percentage of reduction in wound area at 18th day was found to be 98.82% and 73.34% respectively from extract at 400 mg/kg and vehicle control by excision wound model in rat. Granulation tissues weight was 147.24 mg at same dose in incision dead space wound rat model.	Bigoniya et. al, 2007
	Latex	Aqueous	Excision and incision wound model in guinea pig described the extract at 0.5% and 1% sterile aqueous solution facilitated the healing process.	Rasik et. al,   1996
Analgesic	Leaf	Hydro-alcohol	Eddy’s hot plate method increased reaction time. Excellent activity was found against noxious stimuli in tail-flick method.	Gaur et. al, 2009
	Leaf	Hydro-alcohol	Extract at 400 mg/kg showed 432.22% pain inhibition in eddy’s hot plate method (60 minutes). Tail flick and tail clip responses were noted as 416.36 and 165.94% after 45 min at same dose. Same dose also showed protection against acetic acid induced writhing episodes as 53.83%.	Bigoniya and Rana, 2010
Antiulcer	Leaf	Hydro-alcohol	Extract at 200 and 400 mg/kg decreased ulcer index, ulcer grading and free acidity by ethanol-induced ulceration model. Pyloric ligation-induced gastric ulceration model also showed protection at same dose.	Bigoniya and Rana, 2010
Laxative	Leaf	Hydro-alcohol	Extract with castor oil was more potent to produce diarrhea in castor oil-induced diarrheal rat model.	Bigoniya and Rana, 2010
Diuretic	Leaf	Hydro-alcohol	5th hour urine volume was 17.45 ml at 400 mg/kg dose compared to 6.65 ml control in rat model.	Bigoniya and Rana, 2010
Anti-lipidemic	Leaf	Ethanol	Decrease of TC, LDL and VLDL were observed at 400 mg/kg in high fat diet-streptozotocin induced type-2 diabetic rat model.	Mansuri and Patel, 2013
	Leaf	Ethanol,   Petether, Acetone	Extract at 400 mg/kg decreased serum LDL level (4.43±0.16 mg/dl) which was extremely significant compared with the control (46.29±2.12 mg/dl).	Bigoniya andRana, 2009
Hepatotoxicity   & Hepato-protective	Leaf	Ethanol	Significant hepato-protection (54.33%) was formed by 175 mg/kg dose against CCl4-induced hepatotoxicity in rat model. Hiopentone induced sleeping time study showed 85.23% activity. ENSF at same dose and control showed 82.93 and 92.99% viable of liver cell in trypan blue exclusion test.	Bigoniya et. al, 2010
	Leaf	Ethanol,   Pet-ether, Acetone	The extract treatment decreased SGOT due to presence of antioxidants.	Bigoniya andRana, 2009
	Leaf	Hydro-ethanol	Isolated EN flavonoid administration significantly decreased the activities of AST, ALT and ALP.	Sharma and Prachet, 2013
Antispasmodic	Plant	NS	In vitro antiplasmodium assay ensured plant with 2% mercuric chloride has antispasmodic activity.	Kumar N,   2014
Cytotoxicity	Leaf	Ethanol   Pet-ether, Acetone	LD50 of extract was found to be 2779.71 mg/kg.	Bigoniya and Rana, 2009
	Stem	Hydro- alcohol	Extract at different doses up to 2000 mg/kg were observed without toxic reaction or mortality.	Rina et. al, 2013
	Leaf	Ethanol	LD50 of ENSF was found to be 979.24 mg/kg.	Bigoniya et. al, 2010
	NS	NS	Slit-lamp examination revealed a large corneal epithelial defect and moderate stromal edema produced by the extract in human.	Basak et. al, 2009
Mitogenic	Latex	NS	Mitogenic activity test showed lectin possessed activity.	Seshagirirao and Prasad, 1995
CNS depressant	Leaf	Ethanol	Motor co-ordination in mice model showed significant effect.	Bigoniya and Rana, 2005
Psychotropic	Leaf	Ethanol	Elevated plus-maze in mice model showed antianxiety activity and reduced apomorphine induced stereotype. Rat model did not block conditioned or unconditioned response during condition avoidance response.	Bigoniya and Rana, 2005
Anti-convulsant	Leaf	Ethanol	Rat model significantly reduced clonic convulsion phase time and stupor phase.	Bigoniya and Rana, 2005
Antifertility	Stem	Hydro-alcohol	Showed no anti-ovulatory activity. Extract at 400 mg/kg showed 66.66% inhibition of implants in uterine in anti-implantation in rat model.	Rina et. al, 2013
Antidote	NS	aqueous	Performed  like animal or insect-bite antidote	Uawonggul et. al, 2006

 

PHYTOCHEMICAL AND NUTRIENT STUDY

Leaves contain not only primary metabolites including protein, carbohydrate, fat etc. but also diverse secondary metabolites including alkaloids, anthraquinones, glycosides, flavonoids, phenolic compounds, pholobatanins, polyphenol, saponins, steroids, tannins, terpenoids, gums, mucilage, cardenolides etc. (Pokharen et. al 2011; Toume et. al, 2012; Yadav et. al, 2011; Sharma and Pracheta et. al, 2013; Bigoniya et. al, 2010). Alkaloid, flavonoids, glycosides, phenol, lignin, saponins, sterols and tannins etc. several secondary metabolites are found in flower measured by different qualitative tests (Leela et. al, 2013). Methanolic extract of stem contains alkaloids, flavonoids, cardiac glycosides, saponins, tannins and terpenoids (Samaresh et. al, 2013). Terpenoids including antiquorin, neriifolene etc. are found in ethanolic extract of its root (Ng AS, 1989). Latex contains terpenoids including Cycloartenol, 9, 9-cylolanost-20(21) ene-24-ol-3-one (neriifolione) determined by H- NMR, C-NMR, IR and mass spectra. Ammonium sulfate precipitation model, anion exchange chromatography, size-exclusion chromatography analysis showed the presence of Neriifolin S found in latex. Affinity chromatography also determined the presence of lectin (Yadav et. al, 2011; Ilyas et. al, 1998;Seshagirirao and Prasad, 1995). Presence of calcium is also observed in it (Vimala andShoba, 2014; Panda H, 1999). The prominent traditional applications of different parts of it are summarized in Table 2.

PHARMACOGNOSY STUDY

Antibacterial activity

The methanol extract of stem at 400 mg/ml showed highest activity against pseudomonas aeruginosa with inhibition zone of 18 mm where the control streptomycin and ampicillin at 50 μl showed 22 mm. Another result showed inhibition zone 21.32 mm against Staphylococcus aureuswhereas streptomycin showed 4 mm (Samaresh et. al, 2013).The maximum inhibition of chloroform extract at 50µg/ml was observed on P. Vulgarius (8 mm) and E. coli (7 mm). Ethanol extract has shown greater activity against K. Pneumoniae (5.0±0.41mm) and P. Fluoresens (5±1.1 mm) (Pokharen et. al, 2011). Chitosan with latex milk at 60μl formed inhibition zone as 8, 8 and 7 mm against Salmonella typhi, Klebsiella pneumonia and Pseudomonas aeruginosa, respectively (Sumathi et. al, 2012).

Antifungal activity

Antifungal activity of methanol extract showed significant inhibition zone against Aspergillus nigur (50mg/ml-no activity, 100 mg/ml-2 mm, 200 mg/ml-6 mm, 400 mg/ml-12 mm) where the control group of amphotericin B showed 18mm. Candida albicans showed (50 mg/ml-no activity,100 mg/ml-04 mm, 200 mg/ml-10 mm, 400 mg/ml-14 mm) significant level of inhibition zone with the control group 21mm (Samaresh et. al, 2013). Chitosan with latex milk at 60μl reduced the percentage of spore germination in Aspergillusflavus, Aspergillus fumigates and Mucor (Sumathi et. al, 2012).

Antiviral activity

Among of 23 compounds were isolated from ethanolic extract of leaves, 3-β-friedelanol exhibited more potent anti-viral activity than the positive control, actinomycin D implying the importance of the friedelane skeleton as a potential scaffold for developing new anti-HCoV-229E drugs (Chang et. al, 2012).  Eurifoloids E and F exhibited significant anti-HIV activities, with EC50 values of 3.58 ± 0.31 (SI = 8.6) and 7.40 ± 0.94 μM (SI = 10.3), respectively (Zhao et. al, 2014).

Anti-inflammatory activity

Anti-inflammatory effect of hydro-alcoholic extract at 400 mg/kg body weight was observed. Percent inhibition in paw edema was recorded 63.27 % in indomethacin and 55.12 % in extract after 3 hours treatment (Gaur et. al, 2009). Same concentration of extract and aspirin treatment showed 69.47% and 75.78% inhibition of edema volume, respectively. Diclofenac and same dose of extract treatment showed respectively 66.14% and 60.61% inhibition of mass development of granulomatous tissue compared to vehicle control group (Bigoniya and Rana, 2010).

Immuno-modulatory activity

In E. coli induced abdominal sepsis control, control with betamethasone, extract (400 mg/kg) and extract with betamethasone treatment showed 66.6%, 100.0%, 16.6% and 33.3% mortality after 48 hours treatment, respectively. Phagocytic index was found to be 0.0286, 0.0127 and 0.0252, respectively from extract (400mg/kg), control+betamethasone and extract+betamethasone whereas in haem-agglutination antibody titre, primary titres were 14.7±0.26, 5.18±0.20 and 10.2±0.29, respectively and secondary titres were 20.7±0.28, 8.63±0.35 and 14.9±0.28, respectively (Gaur et. al, 2009). Treatment of leaves extract increased the total leukocyte count and differential leukocyte count in normal and immunosuppressed animals ensured it could stimulate the bone marrow activity. The assesement of immunomodulatory activity of hydro-alcoholic extracts of dried leaves (100, 200 and 400 mg/kg) was also evaluated by carbon clearance method, haemagglutination antibody titre method and footpad swelling method on wistar albino rats (Gaur et. al, 2009).

Antioxidant and free radical scavenging ability

Liver lipid peroxidase activity was decreased (7.78±0.69 nmol/mg) after 45 days treatment of extract (400mg/kg) compared with vehicle control (29.94±1.22 nmol/mg). Extract at 200 mg/kg increased the level of SOD 26.06±1.12 unit/ mg in liver, 47.58±2.24 unit/ mg in kidney where the vehicle control group showed only 1.76±0.23 unit/ mg and 4.31±0.69 unit/ mg, respectively after 45 days treatment (Bigoniya and Rana, 2009). The reduction ability of ferric ions found to be 149.2±0.05, 333.1±0 and 43.8±0.06 μmol Fe (ΙΙ)/g, respectively by extract, BHT and quercetin treatment. The percentage inhibition of superoxide generation at 1mg/ml concentration of extract was found to be 50.06%. On the other hand, BHT, BHA, rutin and ascorbic acid at 1mg/ml concentration showed 42.6%, 36.36%, 73.08% and 60.55% inhibition of superoxide radicals respectively. The percentage of metal chelating capacity of extract and standard were found to be 73.24% and 85.37% respectively. At same concentration the scavenging activity of extract reached to 76.15% (Pracheta et. al, 2011). Total sapogenin fraction of leaf extract and α-tocopherol showed 190.36 ± 2.86 and 447.70 ± 6.21 ASE/mg, respectively. The IC50 value was determined from the total sapogenin fraction and α-tocopherol as 440.6 ± 42.3 and 38.6 ± 0.7 µg/ml in hydrogen-donating ability, and 471.3 ±38.2 and 59.6 ± 1.0 µg/ml in superoxide radical production, respectively (Bigoniya and Rana, 2009).  ENSF treatment showed 4.12 ± 0.76 unit/mg enhancement ability of SOD only at 175 mg/kg dose where the control group showed 5.72 ± 0.87 unit/mg. It at same dose showed 14.70 ± 1.22 nmol/g reduction in lipid-peroxidase where the control group and CCl₄ showed 3.43 ± 0.68 nmol/g, 32.97 ± 1.75 nmol/g, respectively (Bigoniya et. al, 2010).

Anti-diabetic activity

Extract with normal control group at 400 mg/kg produced a maximum fall (99.6 ± 2.54 mg/dl) after 60 min glucose administration where the normal control group and glibenclamide (2.5mg/kg) produced 110.2 ± 3.01 and 81.1 ± 2.617 mg/dl, respectively (Mansuri et. al, 2013). Extract at 400 mg/kg and 200 mg/kg showed respectively 43.23±3.58 and 82.48±4.60 mg/dl reduction of glucose where the vehicle control group showed 112.63±4.68 mg/dl after 6 weeks treatment (Bigoniya and Rana, 2009).

Thrombolytic activity

A lectin was purified from latex by affinity chromatography which agglutinated trypsinized human and rabbit erythrocytes, but not sheep erythrocytes (Seshagirirao and Prasad, 1995).

Hematological activity

Oral administration of extract at 200 and 400 mg/kg increased Neutrophil (N) as 41.64±1.49 and 40.52±1.28 %, Lymphocyte (L) as 57.54±1.35 and 57.93±1.31 % where the control group showed as 52.45±1.27 and 45.28±1.94 %, respectively after 21 days treatment. Extract at 200 mg/kg increased Hb, N and L as14.36±0.68 gm/dl, 39.37±1.44 % and 58.54±1.30 %, respectively and at the dose of 400 mg /kg as14.50±0.74 gm/dl, 40.02±1.72 % and 58.86±1.50%, respectively where the vehicle control showed 12.20±0.97 gm/dl, 53.14±1.51 % and 44.17±1.59 %, respectively after 45 days treatment (Bigoniya and Rana, 2009).

Anti-cancer activity

The sapogenin exerted highly significant reduction of gamma radiation-induced chromosomal aberrations (33.5% compared to 71.5% for radiation treatment alone at 4 Gy) which showed IC50 (over a period of 72 hours) of total sapogenin that inhibited growth of mouse melanoma cells by 50% was 173.78µg/ml compared to 120ng/ml for vincristine (Bigoniya and Rana, 2009). Experimental mice were pretreated with 150 and 400 mg/kg body weight of extract, 0.5% and 1% mg/kg body weight of BHA as a standard antioxidant and 50 mg/kg body weight of ENF for 21 days prior to the administration of a single dose of 50 mg/kg body weight of DENA. The study showed significant anti-carcinogenic potential of the extract and ENF against DENA-induced renal carcinogenicity might be due to the presence of flavonoids and phenolic compounds (antioxidants) (Sharma and Pracheta, 2013; Sharma et. al, 2011). Oral administration of hydro-ethanolic extract (150 and 400 mg/kg body weight), control and BHA (0.5 %) significantly decreased LPO level and significantly increased the SOD and CAT  activity (Pracheta et. al, 2011). 3-β-friedelinol, 3-β-taraxerol, 3-α-friedelinol were isolated from extract of the aerial parts of EN showed potent cytotoxic activity on Panc-1, 81T, and BE3 cancer lines, each compound with about 60% inhibition rate at a concentration of 10 µm (Lin et. al, 2013).

Wound healing activity

Epithelization period of methanol extract, control and framycetinwere found to be 13.2 ± 0.48, 17.4 ± 0.81 and 12.5 ± 0.43 days, respectively in excision wound model. Wound healing effect of extract and control on excision wound model was found to be 100.00 ± 1.93 and 85.66 ± 2.78%, respectively at 18^th day (Pattanaik et. al, 2014).  Incision breaking strengths were found to be 700.31 ± 3.67, 389.87 ± 3.86 and 712.23 ± 2.84 g, respectively by extract at 500 mg/kg, standard control and framycetin. Estimation of hydroxyproline content after treatment of extract, standard control and framycetin was found to be 2198.78 ± 1.20, 1401.22 ± 0.98 and 2439.61±0.87μg/100g, respectively (Pattanaik et. al, 2014). Granulation tissue weight was 147.24 mg at 400 mg/kg extract treatment compared to 36.83 mg of control and 165.60 mg of ascorbic acid (Bigoniya et. al, 2007). The wound healing effect of aqueous extract of the latex was evaluated in guinea pig. The 0.5% and 1% sterile aqueous solution of extract facilitated the healing process as evidenced by increase in tensile strength, DNA content, epithelization and angiogenesis (Rasik et. al, 1996).

Analgesic activity

120 minutes treatment of extract showed good analgesic activity by increasing in the reaction time (increased threshold potential of pain)(Gaur et. al, 2009). Pain inhibition was found 432.22% after 60 minutes extract treatment at 400 mg/kg. The extract (400 mg/kg) increased tail flick and tail clip response as 416.36 and 165.94% after 45 minutes treatment. Acetic acid induced writhing episodes (stretch, torson or constriction of abdomen and extension or drawing up of a hind leg) protection was found to be 53.83% at same dose (Bigoniya and Rana, 2010).

Antiulcer activity

Leaf extract decreased ulcer index, ulcer grading and free acidity. Extract at both 200 and 400 mg/kg also decreased significant level of total acidity and volume of gastric content (Bigoniya and Rana, 2010).

Laxative activity

The extract did not produce diarrhoea alone but with castor oil, it produced diarrhoea, which was 20.29 % more than the castor oil alone (Bigoniya and Rana, 2010).

Diuretic activity

Hydro-alcoholic extract of leaves produced dose dependent diuretic activity, which was extremely significant at 400 mg/kg dose whereas increase in total volume of urine formation was significant at both 200 and 400 mg/kg dose. Fifth hour urine volume was 17.45 ml after extract treatment (400 mg/kg) as compared to 6.65 ml of control. The observation also suggested that it was effective on hypernatraemic and hyperchloraemic diuretics (Bigoniya and Rana, 2010).

Hypo and hyper-lipidemic activity

Treatment of extract at 400 mg/kg with diabetic control increased serum HDL cholesterol as 44.07±1.17 mg/dl where the normal control showed 52.85±2.22 mg/dl. Same treatment decreased serum TC, LDL and VLDL as 73.0±1.57, 53.56±1.26 and 35.54±0.99 mg/dl, respectively where the normal control showed 62.84±1.67, 34.03±1.84 and 25.73±1.39, respectively (Mansuri and Patel, 2013).  Extract at 400mg/kg reduced serum lipid profile including cholesterol (as 73.23±3.16 mg/dl), triglyceride (as 42.21±2.76 mg/dl) and LDL (as 4.43±0.16 mg/dl) where the control group showed 104.72±5.17, 65.17±3.07 and 46.29±2.12 mg/dl, respectively after 45 days treatment (Bigoniya and Rana, 2009).

Hepato-protective activity

Silymerin, negative control and vehicle control showed as 87.89, 61.56 and 92.99% viability of liver cell where ENSF at 175 mg/kg showed 82.93% viability of liver cell against CCl₄ induced hepatotoxicity. Liver slices of ENSF treated animals showed extremely statistically significant hepatoprotection (54.33%) at 175 mg/kg dose. It showed 75.44 ±4.12 µg of bromsulphalein uptake per gram of liver tissue compared to 42.11 ±2.38 µg of CCl₄ treated group (Bigoniya et. al, 2010). 45 days treatment of extract at 400 mg/kg and control showed 138.97±5.61 and 68.24±3.84 U/l increasing of ALP. Decrease level of SGOT was 45.55±3.71 and 8.31±0.85 IU/L of vehicle control and extract was also observed (Bigoniya andRana, 2009). Isolated EN flavonoid administration significantly decreased the activities of AST (44.27±0.08 IU/L), ALT (37.82±0.26 IU/L), ALP (64.09±0.29 μmole) and the TP (7.67±0.08 g/ml) content (Sharma and Prachet, 2013).

Antispasmodic activity

Plant has antispasmodic activity if it treated with 2% mercuric chloride solution (Kumar N, 2014).

Cytotoxicity study

LD50 was found to be 2779.71 mg/kg (Bigoniya and Rana, 2009). LD50 of ENSF was determined as 979.24 mg/kg according to the guidelines of OECD following the Up & Down method and fixed dose method (Bigoniya et. al, 2010). Extract at different doses up to 2000 mg/kg were administered and animals were observed for behavioral changes and mortality up to 48 hours providing nontoxic reaction or mortality(Rina et. al, 2013). A lectin was purified from latex by affinity chromatography on sepharose 4B. The Euphorbia lectin possessed mitogenic activity with spleen lymphocytes but it did not inhibit protein synthesis in rabbit reticulocyte lysate (Seshagirirao and Prasad, 1995). Two days exposure of E. neriifolia sap showed large corneal epithelial lesion with stromal edema and mild anterior uveitis (Basak et. al, 2009).

Psychotropic activity

Extract at 400 mg/kg dose exhibited pronounced anti-anxiety activity by increasing the percentage of preference to open arm, open arm entries and time spent in open arm. Leaf extract significantly reduced apomorphine induced stereotype in mice at all the tested dose level compared to apomorphine suggesting the extract might have dopamine receptor modulating activity. Avoidance of conditioned response was examined and rats were trained to climb the pole at the sound of buzzer within 30 seconds. Extra pyramidal effects on rats were measured by placing the animals individually on wooden blocks after test drug and chlorpromazine (3 mg/kg) administration. Leaf extract did not show catatonic effect, as scoring of leaf extract at 200 and 400 mg/kg doses were 0.41±0.11 and 0.82±0.68, respectively in comparison tochlorpromazine 8.58±2.18. Leaf extract caused significant potentiation of scopolamine induced amnesia at 200 and 400 mg/kg on elevated plus-maze employed for the evaluation of short-term memories (Bigoniya and Rana, 2005).

Anti-convulsant activity

Maximum current of 150 mA for 0.2 sec duration was employed on rats using eye electrode of electro convulsiometer. Test drug was given to the animals 1 hour before electro shock and time was taken for each phase observation. Leaf extract at 400 mg/kg dose significantly reduced tonic (both flexor and extensor), clonic convulsion phase time and stupor phase (Bigoniya and Rana, 2005).

CNS Depressant activity

Onset of sleep (loss of righting reflux) was noted and duration of sleep was measured after pentobarbital (45 mg/kg,) administration. Spontaneous motor activity was measured by placing the animals individually in the digital photoactometer for 5 minutes. Fall off time was recorded by placing the animals individually on a rota-rod at 20 rpmas pre-test session. Those animals that stayed on the rod for not less than 3 minutes were selected for the test session (Bigoniya and Rana, 2005).

Antifertility activity

Extract at 200 and 400 mg/kg showed no change in the weight of ovaries and cholesterol level, when compared to vehicle treated group. Both doses of extract showed 16.66% and 66.66 % inhibition of implants in uterine horns when compared with vehicle treated group (Rina et. al, 2013).

Antidote activity

EN extract had a tendency to be scorpion venom antidotes (Uawonggul et.al, 2006).

DISCUSSION

From the above discussion it is clear that Euphorbia neriifolia as a good source of phytochemicals bolsters its ethno botanical significance. Recent pharmacognostical assays also supports its role as a medicinal plant. From prehistoric time, plants were considered not only as a source of food, fodder and fuel but also as a significant remedy for various diseases due to its multifaceted therapeutical values. All over the world, even in the present days of nanotechnology and genetic engineering, traditional medicinal system is prominent because more than three-fourth people are still dependent on traditional medicine. Medicinal plant based research is increasing not only for preventing antibiotic resistance and sudden outbreak of anonymous virus but also for providing comparatively safer natural medicine rather than synthetic drugs. Medicinal value and usage of Euphorbia neriifolia are although well established for thousands of years but huge activities are still left to complete the perfect recognition of the plant. Therefore, this review will be useful to find out the undiscovered potential of this plant.

CONFLICT OF INTERESTS

The authors declare that there is no conflict of interests regarding the publication of this paper.

ACKNOWLEDGMENT

This work was gratefully supported by Department of Biochemistry, School of Science, Primeasia University, Banani, Dhaka, Bangladesh.

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