1] INTRODUCTION:

Hedyotis diffusa Willd (H. diffusa, Family Rubiaceae), known as Oldenlandia diffusa (Willd) Roxb, is a well-known Chinese medicine used for the treatment of inflammation-linked diseases, such as hepatitis, appendicitis and urethritis, for thousands of years in China. Recently, H. diffusa has gained increasing attention for its properties of anti-proliferative activity in cancer cells and anti-tumor activity in tumor-bearing animals.

It has been proved as the most commonly prescribed single Chinese herb used for colon cancer and breast cancer patients. according to the statistics from the medicine in India. It occurs abundantly as a weed throughoutIndia. In India, the plant is used National Health Insurance Research Database of Taiwan.⁽¹⁾is a significant drug of Ayurvedic system of as a medicine with multiple actions such as stomachic, antileprosy, diuretic, antiasthmatic, diaphoretic, anthelmintics, febrifuge, antiscabies and antiurethritis. The ethanolic extract of this planthas exhibited antiproliferative and immunomodulatory properties. The major chemical constituents C- methyl flavones characterized as 5, 7-dihydroxy 3′, 4′-dimethoxy-6, 8-dimethyl flavone, rotenoid analogs, boeravinone A-F have been reported from the roots of B. diffusa.⁽²⁾It is been recorded in history that Medicinal herbs have been used as form of therapy for the relief of pain. The exploration of the chemical constituents from plants, pharmacological and phytochemical screening would provide the basis for developing the new lead molecules in strategic favour of natural product drug discovery.⁽³⁾Oldenlandia corymbosa (L.) Lam. syn Hedyotis corymbosa (L.) Lam. (Rubiaceae) is a weedy annual herb, found especially during monsoon in fields throughout India, Sri Lanka, tropical East Asia to Java and the Phillipines.1 It is commonly known as, Parppatakapullu‟ in traditional medicine of Kerala. The plant is known to clear heat and toxins, activate blood circulation, promote diuresis and relieve stranguria (urinary obstruction). It is also known to act against tumors of the digestive tract lymphosarcoma and carcinoma of the liver and larynx.⁽⁴⁾ Oxygen free radicals produced as a result of metabolic processes in our body, induce damage to biomembranes and genetic materials leading to many chronic degenerative diseases and aging. Oxidative stress, an unbalance between prooxidants and antioxidant mechanisms, directed the use of dietary or medicinal supplements particularly during disease attack.⁽⁵⁾

2] Scientific Name: Linn. Syn. B repens, B. repens⁽³⁾

3] Vernacular names:

Sanskrit: Boerha aviadiffusa, Boerha aviarepens Linn; Punarnava⁽⁶⁾Punarnava, Raktakanda, Shothaghni, Varshabhu⁽³⁾

Hindi: Lal Punarnava, Beshakapore,⁽⁶⁾Snathikari ^(3,6)

English: Spreading Hogweed, Shothagni, Red Hogweed, Raktapunarnava⁽⁶⁾

Malayalam: Thazhuthama.⁽⁶⁾

Bangali: Punarnnava^(3,6)

Kanarese: Kommegida⁽⁶⁾

Gujarati: Vakhakhaparo,Dholia-saturdo^(3,6)

Marathi: Tambadivasu⁽³⁾ Ghetuli^(3,6)

Tamil: Chattarani ⁽⁶⁾Mukaratee-Kirei⁽³⁾

Telugu: Galijeru⁽⁶⁾Punernava,⁽³⁾

Oriya : Lalapuiruni Nalipuruni⁽⁶⁾

Punjabi : ltcit⁽⁶⁾

Ial: Khattan⁽⁶⁾

Kannada: Komma^(3,6)

Kashmiri: Vanjula Punarnava⁽⁶⁾

Assamese: Ranga Punarnabha^(6,7,8)

4] Common Names:

Erva Tostão, Erva Toustao, Pega-pinto, Atikamaamidi, Biskhapra, Djambo, Etiponia, Fowl's Lice, Ganda'dar, Ghetuli, Katkatud, Mahenshi, Mamauri, Ndandalida, OulouniNiabo, Paanbalibis, Patal-jarh, Pitasudu-pala, Punar-nava, Punerva, Punnarnava, Purnoi, Samdelma, San, Sant, Santh, Santi, Satadi Thikedi, Satodi, Spreading, Tellaaku, Thazhuthama, Thikri, Touri-touri, Tshrana, Yoegbe, Beshakapore, Raktpunarnava.⁽⁶⁾ Hog Weed, Pig Weed, Horse purslane, Tarvine⁽⁹⁾

5] Geographical Distribution and Habitat:

It mainly consists 40 species that is distributed in tropical and subtropicalregions and warm climate. It occurs in the Ceylon, Australia, Sudan and Malay Peninsula, extending to China, Africa, America and Islands of the Pacific. Among 40 species of Boerhaavia,⁽¹⁰⁾ species are found in India, namely B. diffusa, B.erecta, B. rependa, B. chinensis, B. hirsute and B. rubicunda. Boerhaavia diffusa is seen in India is found in warmer parts of thecountry and throughout up to 2,000 m altitude in the Himalayanregion. It is a perennial, spreading hogweed, commonlyoccurring abundantly in waste places, ditches and marshy placesduring rains. The plant is also cultivated to some extent in WestBengalOldenlandia corymbosa is native to Africa and India, but now with pantropical distribution; throughout Malaysia.^(3,4,6)

6] Taxonomical Classification:

Botanical Name: Boerha aviadiffusa L.^(6,9)

Family: Nyctaginaceae⁽⁶⁾ rubaceae^(4,9)

SubFamily: rubioidiae ⁽⁴⁾

Kingdom: plantae⁽⁴⁾

Phylum: angiosperms⁽⁴⁾

Class: Magnoliopsida⁽⁶⁾ dicotyledeonae⁽⁴⁾

Subclass: aasteridaen⁽⁴⁾

Division: Magnoliophyta⁽⁶⁾

Order: Caryophyllales⁽⁶⁾gealentianes⁽⁴⁾

Genus: Boerhaavia ⁽⁶⁾oldenlandia^(4,9)

Species: B. diffusa⁽⁶⁾ corymbosa^(4,9)

7] Plant Description:

Stem:

It is greenish purple in colour, stiff, slender, cylindrical, swollen at the nodes ends, minutely pubescent, prostrate divaricately branched, often more than a meter long. Ascending, 4-10 cm long.⁽⁶⁾

Root:

Roots are well developed, fairly long, somewhat tortuous, cylindrical, 0.2-1.5 cm in diameter, yellowish brown to brown in the colour, surface soft to touch but rough due to minute longitudinal striations and root scars, short, no distinct odour, slightly bitter, sweet, pungent.⁽⁶⁾The roots are stout and fusiform with a woody root stock. It has a large root system bearing rootlets. Thetap root is tuberous, cylindrical to narrowly fusiform to conical⁽³⁾It is thick. These roots contained 0.15% alkaloid punarnavine. Increase in levels of indole- acetic acid (IAA) in MS medium reduced the number of roots regenerated from the leaf segment, their length and alkaloid content⁽⁹⁾

Leaves:

Opposite in unequal pairs, larger ones 25-37 mm long and smaller ones 12-18 mm long ovate-oblong or suborbicular, apex rounded or slightly pointed, base subcordate or rounded, green and glabrous above, whitish below, margin entire or sub-undulate, dorsal side pinkish in certain cases, thick in texture, petioles nearly as long as the blade, slender.⁽⁶⁾The shape of the leaves varies considerably - ovate-oblong, round, or subcordate at thebase and smooth above. Margins of the leaves are smooth, wavy, or undulate. The upper surface of theleaves is green, smooth, and glabrous, whereas it is pinkish white and hairy beneath. Leaves are up to 5.5 ×3.3 cm2 in area.⁽⁹⁾

Flowers:

Flowers are minutely small, pink coloured nearly sessile or shortly stalked, 10-25 cm, in small umbrellas, arranged on slender long stalks, 4-10 corymb, axillary and in terminal panicles, bracteoles, small, acute, perianth tube constricted above the ovary, lower part greenish, ovoid, ribbed, upper part pink, funnel-shaped, 3 mm long, tube 5 lobed, stamen 2-3.⁽⁶⁾. Usually fascicledor subumbellate on the ultimate branchlets, pink, andabout 1.5 mm long. A perianth is present in the place of acalyx and corolla, which is tubular in shape, the tubebeing short and narrow at the base and funnel-shaped atthe top and constricted above the ovary. There are fivelobes, which are small and acute. Two or three stamensare present and are slightly exserted. The stigma ispeltate.⁽⁹⁾In pendunculate, glomerulate clusters arranged in slender, long stalked, axillary or terminal corymbs

Fruit:

One seeded nut, 6 mm long clavate, rounded, broadly and bluntly 5 ribbed, viscidly glandular.⁽⁶⁾The achene fruit is detachable, ovate, oblong, pubescent, five-ribbed and glandular, anthocarpous, andviscid on the ribs. Fruit is glandular, narroly oblongobovoid, about 3 mm long. about 3 mm long. These are hermaphrodite, pedicellate, and white, pink, or pinkish-red in color. Bracts are deciduous and involucrate.⁽⁹⁾Fruiting Throughout the year in Indian conditions Ovoid or sub-ellipsoid, rounded above, slightly cuneate, below, broadly and bluntly 5-ribbed, very glandular throughout⁽³⁾

9] Pharmacological actions:

Antibacterial Activity:

A Potent antibacterial activity against gram positive and gram negative bacteria shown by the leaves of B. diffusa are due to the phytochemicals present in the leaves.⁽¹²⁾The antibacterial activity of the various extracts of the stem bark of Prosopis cineraria (Linn.) Druce, was evaluated by the agar well diffusion method⁽¹³⁾The aqueous and ethanolic extracts of B. diffusa leaves had activity on E. coli, S. aureus and P. aeruginosa⁽¹⁴⁾

Analgesic/Anti-Inflammatory Activity:

The Decoction (DE) or Juice (JE) of the leaves of Boerhaavia diffusa were used to study the antinociceptive effect in chemical (acetic acid) and thermal (hot Plate) models of hyperalgesia in Mice. The DE, raised the pain thresholds during the first period (30 min) of observation.⁽¹⁵⁾The analgesic property of B. diffusa was mainly showed by its leaf part. The mechanism of its analgesic effect was not known. The study proved that the active anti-nociceptive principle of B. diffusa was present mainly in the aqueous extract of fresh leaf juice.⁽¹⁶⁾

Anti-Convulsant Activity:

The study was carrired out to investigate the methanolic root extract of B. diffusa and its different fractions including liriodendrin-rich fraction for exploring the possible role ofliriodendrin in its anti-convulsant activity.⁽¹⁷⁾The methanolic extract of root part of B.diffusa was reported for anticonvulsant activity in PTZ induced convulsion in mice⁽¹⁸⁾

Antidiabetic Activity:

The study indicates that Boerhaavia diffusa and ethanolic extracts exhibit significant antihyperglycemic activities in alloxan induced as well asstreptazotocin induced hyperglycemic rats. Similar effects in streptozotocin-treated diabetic animals were reported by pancreastonic, ephedrine, and Gymnema sylvestre leaf extracts. In the current studies, the damage of pancreas in streptazotocintreated diabetic control rats and regeneration of cells by glibenclamide was observed.⁽¹⁹⁾

Antistress/Adaptogenic/Immunomodulatory Activity:

Hydroethanolic extract (80%) of Boerhaavia diffusa (HEBD) and a polyherbal formulation (Punarnava mandur) PHF-09 containing Boerhaavia diffusa were compared for their antistress activity using cold restraint stress model. Stress was induced by subjecting animals to cold restraint⁽²⁰⁾

Antitumor Activity:

Cancer chemo preventive property of B. diffusa was evaluated on 7,12-dimethyl benz (a) anthracene (DMBA) induced skin papillomagenesis in male Swiss albino mice (6-7 weeks old). The cancer chemopreventive efficacy was assessed by its ability to modulate the activities of enzymes associated with drug metabolism and bifunctional modulators reduced the availability of ultimate carcinogen metabolites in the epithelial stage ⁽²¹⁾

Antiproliferative and Antiestrogenic Activity:

Antiproliferative and antiestrogenic properties of methanol extract of Boerhaavia diffusa (BME) in MCF-7 breast cancer cell lines. Boerhaavia diffusa extracts exhibited a strong inhibitory effect on the proliferation of human breast cancer cells in vitro and the antiestrogenic effects are mediated by ER. Phytochemical studies have revealed the presence of alkaloids, flavonoids, phenols and saponins in BME. The antiestrogenic activity shown by the extract may be attributed to these diver compounds⁽²²⁾

Anti Fibrinolytic Activity:

A study evaluated the effect of antifibrinolytic agents, _-aminocaproic acid (_-ACA), tranexamic acid (AMCA); anti-inflammatory drugs (indomethacin, ibuprofen, naproxen), and plant extract (root extract of Boerhaavia diffusa) on endometrial histology of IUD-fitted menstruating monkeys. It is effective in reducing stromal edema, inflammation, and tortuosity of glands, and in increasing the degree of deposition of fibrin and platelets in the vessel lumen⁽²³⁾

Antioxidant Activity:

The evaluation of the antioxidant potential of ethanolic extract of Andrographis echioides and Boerhavia diffusa was carried out by determining the levels of enzymatic and non-enzymatic antioxidants.⁽²⁴⁾ The roots of B.diffusa were mentioned by researchers for their potential antioxidant property Where as ethanolic root extract of this plant was reported for its potent antioxidant activity on enzymatic and nonenzymatic experiments⁽²⁵⁾

Antiviral Activity:

Boerhaavia diffusa has many ethnobotanical uses (the leaves are used as vegetable; the root juice is used to cure asthma, urinary disorders, leucorrhoea, rheumatism, and encephalitis), and is medicinally used in the traditional, Ayurvedic system. Besides, the Boerhaavia Diffusa plant is reported to possess many pharmacological, clinical, and antimicrobial properties.⁽³⁾

Acute oral toxicity test:

The toxicity test of aqueous extract of Oldenlandia corymbosa L. was done to determine the safe dose for oral consumption. The study was performed in five groups of six mice. Thirty male and non pregnant female Swiss albino mice weighing 20grams each were used for this study. They were divided into the Control (C) and Test (T) groups. Each group except the control was further divided into four sub-groups of six mice. The aqueous extract of Oldenlandia corymbosa was administered orally at the dose of 5, 10, 20 and 40 g/kg body weight after 12 hours fasting. LD50 was obtained using Thompson and Finney method. LD50 of aqueous extract of Oldenlandia corymbosa was 14.14+/- 0.27 and 10.56 +/- 0.20⁽²⁶⁾

Anti-malerial activity:

Methanolic extracts of Oldenlandia corymbosa was tested in vitro on choloroquine sensitive (MRC-pf-20) and resistant (MRC-pf-303) strains of Plasmodium falciparum for its anti-malarial activity. Growth inhibition was determined using different concentrations of plant extracts on synchronized P. falciparum cultures at the ring stage. The 50% inhibitory concentration (IC50) of Oldenlandia corymbosa was found 10.8 μg/ml. Combination of Oldenlandia corymbosa and Andrographis paniculata showed substantial enhancement in their anti-malarial activity.⁽²⁷⁾

Abortifacient activity:

Cumulative doses of plant extracts were added to rat uterine tissue in an organ bath, and the force and frequency of contractions were recorded. Acetylcholine was used as positive control. The strongest contraction comparable to the maximum response obtained with acetylcholine was observed in Oldenlandia corymbosa plant extract.⁽²⁸⁾

Adaptogenic activity/Antistress activity:

A polyherbal formulation and hydroethanolic extract of B. diffusa were reported for their antistress activity by resulting in significant decrease in triglyceride levels 43. It is believed that the presence of certain phytochemicals like flavonoids, alkaloids, glycosides and sterol in the ethanolic roots extract of B. diffusa could be responsible for its antistress activity by researchers⁽²⁹⁾

Anti-fertility:

The aqueous ethanolic extract of root of B. diffusa was reported for its anti-fertility activity⁽³⁰⁾

Antilymphoproliferative Activity:

It inhibited T cell mitogen phytohemagglutinin and concanavalin A-stimulated proliferation of human peripheral blood mononuclear cells (PBMC). It also inhibited purified protein derivative antigen-stimulated PBMC proliferation and human mixed lymphocyte culture.⁽³¹⁾

Cytological Activity:

Ethanolic extract of Oldenlandia corymbosa (EOC) was examined for its cytotoxic activity and its effect in combination with doxorubicin against MCF-7 breast cancer cell line as cell model of doxorubicin resistance. Therefore EOC has a potential activity to be developed as co- chemotherapeutic agent.

The plant B. diffusa was reported by researchers for its cytological activity. The root extracts of B. diffusa was mentioned for inhibiting mitosis and also reported for cytological studies over expensive colchicines⁽³²⁾

Hepatoprotective Activity:

The hepatoprotective activity of roots of different diameters were collected in three seasons, rainy, summer and winter, and examined in thioacetamide intoxicated rats⁽³³⁾The extract also produced an increase in normal bile flow in rats suggesting a strong choleretic activity. The extract does not show any signs of toxicity up to an oral dose of 2 g/kg in mice⁽³⁴⁾

Nitric Oxide Scavenging Activity:

The extracts of various polyherbal drugs exhibited dose-dependent no scavenging activities and the potency was in the following order: abana>chyavanaprasha> triphala>geriforte>septilin> mentat >Gingko biloba.⁽³⁵⁾

10] Chemical Test:

1. Flavonoids:

a) Alkaline reagent test:

2ml of the extract was taken and treated with sodium hydroxide solution. Immediately the appearance of yellow or red color confirms the presence of flavonoids.

2. Saponins:

a) Froth test:

1ml extract was taken and 20ml of distilled water was added to it. The mixture was shaked for a minute. Presence on the top confirms the presence of saponins.

3. Proteins:

a) Ninhydrin’s test:

2ml of the extract was taken and few drops of ninhydrin reagent was added to the extract solution and the mixture was heated for some time in a water boiling bath. Appearance of blue color reveals the presence of proteins.

b) Lowry’s test:

2ml of the extract was taken and it was added with 1ml of lowry’s reagent (alkaline copper reagent) and incubated at room temperature for 10mins. Then 1ml of folin phenol reagent was added and again the mixtures were incubated for 30mins. Appearance of blue color complex denotes the presence of proteins.

c) Bradford’s test:

2ml of the bradford’ s reagent was added to 1ml of the extract and it was incubated for 5 minutes at room temperature. Formation of blue color confirms the presence of proteins.

4. Carbohydrate Test:

a) Molisch’s reagent / Napthol test:

2ml of the extract was taken and 1ml of the Molisch’s reagent was added to it. Then 1ml of concentrated sulfuric acid was added along the sides of the test tube. On the appearance on a violet / purple ring at the junction of 2 liquids confirms the presence of carbohydrates.

b) Fehling’s test:

2ml of the extract was taken and 1 ml of Fehling’s solution A and 1ml of Fehling’s solution B was added to the extract. Then the mixture was boiled in a boiling water bath for 2-3 minutes. Formation of brownish red / yellow precipitate confirms the presence of carbohydrates.

c) Benedict’s test:

2ml of the extract was taken and few drops of Benedict’s reagent was added to the extract and it was boiled for 2minutes in the water bath. Later it was allowed to cool. Formation of red, yellow or green color precipitate confirms the presence of carbohydrates.

5. Quinines:

a) Sulfuric acid test:

2ml of the extract was taken and 1ml of the concentrated sulfuric acid was added and left undisturbed for few minutes. Development of red color shows the presence of quinines.

6. Phenols:

a) Ferric chloride test: A few drops of ferric chloride was added with few drops of the extract. Appearance of blue-green color denotes the presence of phenols.

7. Alkaloids:

a) Wagners reagent test:

2ml of the extract was taken and few drops of Wagners reagent was added to it. Formation of red color indicated the presence of alkaloids.

8. Glycoside:

a) Keller Killiani test:

Few drops of the extract was dissolved in acetic acid containing few amount of ferric chloride. The sulfuric acid was added to the test tube. At the junction formation of reddish brown color, which turns blue gradually, confirms the presence of glycoside.

9. Isoflavanoids:

2ml of the extract was taken and treated with sodium hydroxide solution. Immediately the appearance of yellow or red color can be observed. When a few drops of dil. hydrochloric acid was added, the solution turns colourless. This confirms the presence of isoflavinoids.⁽⁸⁾

Traditional Uses:

Each part has a different therapeutic value and must be prepared in its own way for maximum benefits. This plant rejuvenates liver, male reproductive system and other organ system, detoxifies liver and skin; aphrodisiac; increases libido, erection and quality and quantity of semen; reduces cough, asthma etc. It is used in Vajikarana preparations. This plant cleanses the kidneys and helps to get rid of renal calculi (kidney stones). Mainly, the roots and the whole plant is used for the medicinal purpose, externally punarnava is used for alleviate the pain and swelling. The fresh juice of its roots instilled into eyes, mitigates the ailments of the eyes like night blindness and conjunctivitis. The paste applied on the wounds, dries up the oozing. Internally, punarnava is beneficial to treat a wide range of diseases. Punarnava is the most commonly used and the best herb to alleviate swelling, due to its potent diuretic property. It boosts up the filtration, rejuvenates the renal functions and takes out the excessive fluids and Kelda by augmenting the urinary output. The sesame oil, medicated with punarnava is very useful as an adjunct to operating enemas in the treatment of ascites of vata type aflatulence. In large doses, punarnava acts as a purgative. Punarnava effectively reduces fever, especially in malaria. The decoction of rasna, sunthi and punarnava is the best panacea for rheumatic swollen joints, as rasna alleviates the pain and vata, sunthi destroys ama and punarnava reduces the swelling. Punarnava enhances the quality of 6 of the 7 categories of bodily tissues, including nutrient plasma (Rasa Dhatu), blood (RaktaDhatu), muscle (MamsaDhatu), fat (MedaDhatu), bone marrow and nerves (MajjaDhatu), and reproductive fluids (ShukraDhatu).⁽⁶⁾

CONCLUSION:

Oldenlandia corymbosa is an interesting example of a plant used in traditional medicine for many years and the reported phytochemical and pharmacological studies supports its traditional uses and proved to be useful for clinical commercial studies. The multiple benefits of Boerhaavia diffusa made it a true miracle of nature. Numerous studies have been conducted on different parts of Boerhaavia diffusa plant has not yet developed as a drug by pharmaceutical industries. A detailed and systematic study is required for identification, cataloguing and documentation of plants, which may provide a meaningful way for the promotion of the traditional knowledge of the herbal medicinal plants. In view of the nature of the plant, more research work can be done on humans so that a drug with multifarious effects will be available in the future market.

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Received on 15.07.2019 Modified on 31.07.2019

Res. J. Pharmacognosy and Phytochem. 2019; 11(3):137-144.

DOI: 10.5958/0975-4385.2019.00023.2

Sr. No	Chemical Parametre	Summer	Monsoon	Winter
01	Total Ash Content	11.09±0.81	11.66±0.63	11.57±0.54
02	Acid Insoluble Ash content	2.78±0.52	2.39±0.59	2.99±0.83
03	Water Soluble Ash content	6.35±0.65	6.38±0.34	6.95±0.91
04	Sulphated Ash content	15.72±0.87	16.40±0.34	15.26±0.74
05	Ethanol soluble extractive	6.41±0.83	6.7±0.12	6.12±0.31
06	Water soluble extractive	12.96±0.27	13.16±0.20	12.68±0.33
07	Moisture content	5.24 ±0.19	5.79 ±0.53	5.93 ±0.43