Chemotaxonomy of the ethnic antidote Aristolochia indica for aristolochic acid content: Implications of anti-phospholipase activity and genotoxicity study.

ETHNOPHARMACOLOGICAL RELEVANCE: Aristolochia indica L. (Aristolochiaceae) is a common medicinal plant described in many traditional medicine as well as in Ayurveda used against snakebites. Besides, the plant has also been reported traditionally against fever, rheumatic arthritis, madness, liver ailments, dyspepsia, oedema, leishmaniasis, leprosy, dysmenorrhoea, sexual diseases etc. The plant is known to contain its major bioactive constituent aristolochic acid (AA) known for its anti-snake venom, abortifacient, antimicrobial and antioxidant properties. MATERIALS AND METHODS: This present work describes a validated, fast and reproducible high performance thin layer chromatography (HPTLC) method to estimate AA from the roots of 20 chemotypes of A. indica procured from 20 diverse geographical locations from the state of West Bengal, India. Further, an evidence-based approach was adopted to investigate the reported anti-venom activity of the aqueous extracts of the A. indica roots by assessing its phospholipase A2 (PLA2) inhibitory properties since PLA2 is a major component of many snake-venoms. Finally, the cytotoxicity and genotoxicity of the aqueous root extract of the Purulia (AI 1) chemotype were assessed at various concentrations using Allium cepa root meristematic cells. RESULTS: The highest amount of AA (7643.67 µg/g) was determined in the roots of A. indica chemotype collected from Purulia district followed by the chemotypes collected from Murshidabad, Jalpaiguri and Birbhum districts (7398.34, 7345.09 and 6809.97 µg/g respectively). This study not only determines AA in the plants to select pharmacologically elite chemotypes of A. indica, but it also identifies high AA producing A. indica for further domestication and propagation of the plants for pharmacological and industrial applications. The method was validated via analyzing inter-day and intra-day precision, repeatability, reproducibility, instrumental precision, limit of detection (LOD) and limit of quantification (LOQ) and specificity. Chemotypes with high AA content exhibited superior anti-PLA2 activity by selectively inhibiting human-group PLA2. Moreover, A. indica root extract significantly inhibited mitosis in Allium cepa root tips as a potent clastogen. CONCLUSIONS: The present quick, reproducible and validated HPTLC method provides an easy tool to determine AA in natural A. indica plant populations as well as in food and dietary supplements, a potential antivenin at one hand and a possible cause of aristolochic acid nephropathy (AAN) at another. Besides, the cytotoxic and mitotoxic properties of the root extracts should be used with caution especially for oral administration.

The effect of diamino diphenyl sulfone on the embryonic development of eggs from the sea urchin (Lytechinus variegatus).

Diamino diphenyl sulfone (DDS), a chemotherapeutic compound used in the treatment of leprosy, was studied with regard to its effects on the embryonic development of eggs from the sea urchin Lytechinus variegatus. DDS disturbs the normal development of fertilized eggs by interfering with the mitotic apparatus and producing spheroid-shaped bodies shown to be aster-like bodies by staining techniques. A typical colchicine effect (formation of kidney-shaped cells) was also demonstrated.

In vitro effect of dapsone on human chromosomes.

The proportion of accumulated metaphases, as well as the frequency of aneuploidies and structural chromosome aberrations were investigated in DDS-treated and untreated (controls) leukocyte cultures from two samples of healthy adult Caucasoid individuals. In one sample, the sulfone-treated cultures differed from the controls in that they contained 0.4 mug of DDS per ml of tissue culture medium plus phytohemagglutinin. In the other sample, these cultures differed from the controls in that they contained a tenfold higher concentration of DDS. The two concentrations of DDS used have not significantly affected the rate of metaphases, while 0.4 mug/ml DDS did not increase the frequency of chromosome aberrations. The same was not true for the cultures treated with 4 mug/ml DDS in which the proportion of aneuploides and achromatic gaps increased significantly.