The ethanolic extracts from fresh apical stems of Phyllanthus niruri L. (Euphorbiaceae) cultured on Murashige and Skoog (MS) medium supplemented with IBA/BAP/Coco nucifera L. milk for 1, 2, 4 and 6 months were phytochemically and biologically investigated and compared with intact plant part and whole plant extracts. Results from the in vitro antiplasmodial testing indicated that the EtOH extract of a 1-month-old callus culture (IC(50) = 16.3 +/- 2.5 microg/ml) exhibited a higher activity than the ethanolic extracts of the fresh apical stem (IC(50) = 18.2 +/- 2.4 microg/ml) and callus cultures of 2-, 4- and 6-months-old (25 microg/ml < IC(50) < 40 microg/ml). These activities were however lower than that displayed by the ethanolic extract of the whole plant (IC(50) < 3 microg/ml). The EtOH extract of 1-month-old callus culture (the most active) was fractionated with solvents of different polarities. Its CH(2)Cl(2) fraction rich in terpenic constituents (IC(50) = 9.2 +/- 3.4 microg/ml) exhibited a higher antiplasmodial activity than its isoamylic alcohol fraction obtained at pH 2-3 (IC(50) = 25.6 +/- 2.3 microg/ml) rich in flavonoids. The activity of these two fractions was lower than that displayed by the same fractions from the whole plant (2 microg/ml < IC(50) < 3 microg/ml). Alkaloidic fractions from the whole plant and 1-month-old callus culture of fresh apical stem were considered as inactive (IC(50) > 100 microg/ml).
"Traditional medicine using plant extracts continues to provide health coverage for over 80% of the world's population, especially in the developing world (Igbinosa et al., 2009). In the Democratic Republic of Congo (DRC), among the species used in the treatment against malaria, Phyllanthus niruri is well positioned for different previous studies on this plant (Pauwels, 1993; Tona et al., 1999; Cimanga et al., 2004). P. niruri is one of the most important medicinal plants used in different regions in the world for the treatment of various diseases such as jaundice, asthma, hepatitis, flu, dropsy, diabetes, fever causing by malaria (Kerharo and Adam, 1974; Ishimari et al., 1999; Paranjape , 2001) but its availability is drastically decreasing because of numerous harvests. "
"In vitro multiple shoots induction in Phyllanthus amarus had been achieved on Murashige and Skoog (MS) 1962 basal medium (MS) modified with sucrose and low concentrations of benzyl amino purine (0.1 mg/L BAP) and naphthalene acetic acid (0.05 mg/L NAA (Ghanti et al., 2004). Also, calli induction in several species of Phyllanthus has been reported by several authors like Catapan et al. (2001), Cimanga et al. (2004) and Luyindula et al. (2004). Linyindula et al. (2004) successfully induced calli from apical shoots of P. niruri on a medium supplemented with 4 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D). "
[Show abstract][Hide abstract] ABSTRACT: We investigated the use of in vitro techniques for regeneration of adventitious shoots in Phyllanthus niruri, an anti-malarial plant as an initial effort towards its domestication. Fruits cultured on Murashige and Skoog (1962) basal medium (MS) supplemented with BAP had low germination (61%) due to seed coat imposed dormancy. The culture of nodal cuttings explants on BAP, kinetin or 2-isopentyl adenine (2iP) amended medium to avoid seed dormancy resulted in shoot regeneration without roots in all accessions with BAP producing the highest number of shoots (9.0). Subsequent inclusion of either 1-naphthalene acetic acid (NAA) or indole-3-butyric acid (IBA) in the BAP, kinetin or 2iP amended MS medium also produced only shoots. Leaf lobe explants cultured on only 2,4-dichlorophenoxy acetic acid (2,4-D) amended medium led to a significant calli development with 1 mg/L 2,4-D producing 100, 88.9 and 95.8% callus, respectively from Kwabenya, Kasoa and Aburi accessions. Subsequent transfer of calli to MS medium supplemented with 0.1 mg/L BAP led to calli growth (increase in weight) and morphogenic response depending on the concentration of 2,4-D in the induction medium. Only 55 and 25% of these calli from Kwabenya and Kasoa, respectively produced shoots while roots development was significantly higher ranging from 48 to 88.9%. These shoots did not survive ex-vitro acclimatisation due to hyperhydricity while those regenerated from nodal cuttings or seeds had high percentage survival. The high morphogenetic response of Phyllanthus niruri in vitro can be used to propagate this anti malarial plant and enhance its utilization in the treatment of malaria.
"Phyllanthus niruri, a medicinal plant found in the tropics and other parts of the world, exhibits the ability to block the formation of calcium oxalate crystals (Campos and Schor, 1999; Freitas et al., 2002) and stone formation in urolithiasis (Barros et al., 2003, 2006). Furthermore, it is used to treat hyperglycemia (Raphael et al., 2002), hypertension (Srividya and Periwal, 1995), pain (Miguel et al., 1996), and mild malaria (Tona et al., 2001; Cimanga et al., 2004; Mustofa et al., 2007). P. niruri is said to protect against hepatitis B virus (Mehrotra et al., 1990), chemical toxins (Lee at al., 2006; Chatterjee et al., 2006), liver cancer (Rajeshkumar and Kuttan, 2000), and tumorigenesis (Rajeshkumar et al., 2002). "
[Show abstract][Hide abstract] ABSTRACT: Phyllanthus niruri is a medicinal plant (commonly known as stone breaker) found in the tropics and other parts of the world. It is known for its capacity to block the formation of calcium oxalate crystals and kidney stone formation in urolithiasis. This plant has been used to treat hyperglycemia, hypertension, pain, and mild cases of malaria. We examined the geno-, cyto- and overall toxicity of P. niruri whole plant ethanolic extract. The extract was administered as a single dose of 30 or 300 mg/kg to laboratory rats by gavage, accompanied by negative (0.9% saline) and positive (10 mg/mL N-ethyl-N-nitrosourea) controls that were injected intramuscularly 48 h after extract administration. The ratio of polychromatic (PCE)/normochromatic erythrocytes (NCE) from femur bone marrow was scored for genotoxicity. Cytotoxicity was determined using descending concentrations (0.2-0.0125 g/mL) of the extract incubated with peripheral blood mononuclear cells. Lactate dehydrogenase release from damaged cells was determined and the CC(50) calculated. Subchronic administration of the extract at 30 or 300 mg/kg was done for 90 days to determine general toxicity. PCE:NCE (%) for the extract and negative control was 63, compared to 168 (positive control). The CC(50) was 26.3 mg/mL and hepato-renal toxicity after subchronic extract administration was nil. We conclude that ethanol extract of P. niruri is not cytotoxic or genotoxic, and is generally non-toxic on subchronic administration.
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