NUTRITIONAL, PHYTOCHEMICAL AND PHARMACOLOGICAL INVESTIGATION OF BOERHAVIA PROCUMBENS

Abstract

Boerhavia procumbens is an herbaceous, perennial creeping weed and native of Pakistan, belongs to the family Nyctaginaceae. In the folk medicine B. procumbens has been used in the treatment of different diseases containing jaundice, hepatitis, and asthma. This research work was design to investigate the crude methanolic extract, its various solvents soluble fractions and isolated compounds of B. procumbens for biological activities (in-vitro and in-vivo) along with chemical composition to provide a valid scientific rationale to its ethno-medicinal uses. In the proximate analysis, the protein (37.46 ± 0.02 %) and inorganic matter (21.25 ± 0.032 %) of the roots were significantly higher (p < 0.05) than the aerial parts (2.18 ± 0.04 %, 6.78 ± 0.05). The highest value of Nitrogen-free extract (75.56 ± 0.046 %) was found in aerial parts as compared to roots (31.22 ± 0.05 %). Calcium and potassium were significantly highest (p<0.05) elements in the inorganic matter analysis of aerial parts than roots. The essential amino acid, arginine (8.89 ± 0.08 g/100g) and tryptophan (3.49 ± 0.08 g/100g) was found significantly higher (p < 0.05) in roots than aerial parts while non-essential amino acid, aspartic acid (31.25 ± 0.08 g/100g) and glutamic acid (25.27 ± 0.06 g/100g) were reported higher in aerial parts than roots of B. procumbens. Qualitative phytochemical screening of B. Procumbens showed that phenols, flavonoids and cardiac glycoside were present in crude methanolic extract while only phenolic and flavonoidal contents were determined quantitatively in extract and its different solvent fractions. The EtOAc solvent fraction showed significant DPPH radical scavenging activity (IC50: 40.24 μg/mL) when compared to standard (Ascorbic acid, IC50: 28.78μg/ml). The antibacterial activity of dichloremethane (DCM) solvent fraction (12 ± 0.12 mm) showed good zone of inhibition against Xanthomonas campestris while ethyl acetate fraction exhibited maximum activity against E.coli (11.8 ± 0.14 mm). In the antifungal activity only n-hexane fraction (23.2 ± 0.20 mm) showed comparable inhibition against Fusarium oxysporum to that of standard (23.6 ± 0.16 mm). The DCM fraction (90 %) exhibited a highest cytotoxicity activity against brine shrimp at high concentration (1000 μg/mL) whereas, moderate cytotoxic effect (60 and 10 %) was observed at lower concentrations (100 and 10 μg/mL), respectively. The larvicidal effect against 3rd instar larvae of Culex quinquefasciatus of leaf extract and n-hexane fraction showed highest percent mortality (100 % and 96 %) at high concentration (400 μg /ml) with LD50 value of 26.84 and 46.98 μg /ml. The DCM fraction showed outstanding phytotoxic potential (50, 63, and 80 %) while n-hexane fraction showed mild activity (13, 20 and 26 %) at the test concentrations of 10, 100 and 1000 μg/mL respectively. In this study six compounds {Indole-3-carboxaldehyde (1), 2- Hydroxybenoic acid (Salicylic acid) (2), 4-Hydroxybenzoic acid (3), Oleracein E (4), Methyl-3, 4-dihydroxybenzoate (5) and Eupalitin-3-O-β-D-galactopyranoside (6)} were isolated and characterized for the first time from B. procumbens. Compounds (5) and (4) showed a significant antileishmanial activity with IC50 values of 18.37 ± 0.07 ii μM and 48.19 ± 0.06 μM while compounds (4) and (6) depicted good inhibition against acetylcholinesterase enzyme with IC50 value of 54.39 ± 0.03 μM and 74.07 ± 0.06 μM. In Treg cell proliferation assay, Compound (4) was found most active against treg cell proliferation with percent increase (83.51) followed by compound 1 and 6 with percent increase (32.02 and 24.18), respectively. In the isolated compounds only Compound 4 showed less cytotoxicity against NIH-3T3 cell lines with IC50 value of 38.81 ± 2.93 at the test concentration of 100 μM. In-vivo biological activities, toxicity test of BP showed no sign of toxicity and any abnormal behavior up to a dose of 2000 mg/kg for crude extract while for compound (6) up to a dose of 500 mg/kg. The peripheral analgesic activity of crude extract of B. Procumbens at dose of 200 mg/kg exhibited significant effect (p<0.05) while at dose of 400 mg/kg showed more significant effect (p<0.01) against the antinociceptive activity. The central analgesic activity of extract (400 mg/kg) showed more significant effect (p<0.01) after 60 min of administration and remained significant till 120 min. The anti-inflammatory activity of crude extract (400 mg/kg) showed maximum percent inhibition (47.98 %) of paw edema at 5 hrs of observation. In hepatoprotective assay, compound (6) of B. Procumbens at dose of 60 mg/kg with CCl4 significantly reduced (p<0.01) the levels of SGPT and SGOT (26 ± 1.34 U/ml and 42.92 ± 1.6 U/ml) enzymes when compared with the group treated with CCl4 alone (23.85 ± 1.72 U/ml and 34.53 ± 2.41U/ml). Similarly, the serum levels of ALP and total bilirubin (179.22 ± 3.41U/ml and 3.23 ± 0.19 mg/dl) significantly decline (p < 0.01) was also observed in group treated with 60 mg/kg dose. The current research work strongly supports the ethnomedicinal properties of B. procumbens which has been used widely without scientific validation for the treatment of many diseases especially hepatic disorders. Therefore, B. Procumbens can utilizing for value added herbal medicines/products as this species grow commonly as weeds in Pakistan even under tough conditions of environment.