J. R. Vedasiromoni

Curcumin, the major component of food flavour turmeric, reduces mucosal injury in trinitrobenzene sulphonic acid-induced colitis.

Inflammmatory bowel disease (IBD) is characterized by oxidative and nitrosative stress, leucocyte infiltration and upregulation of proinflammatory cytokines. In this study, we have investigated the protective effects of curcumin, an anti‐inflammatory and antioxidant food derivative, on 2,4,6‐ trinitrobenzene sulphonic acid‐induced colitis in mice, a model for IBD. Intestinal lesions (judged by macroscopic and histological score) were associated with neutrophil infiltration (measured as increase in myeloperoxidase activity in the mucosa), increased serine protease activity (may be involved in the degradation of colonic tissue) and high levels of malondialdehyde (an indicator of lipid peroxidation). Dose–response studies revealed that pretreatment of mice with curcumin (50 mg kg−1 daily i.g. for 10 days) significantly ameliorated the appearance of diarrhoea and the disruption of colonic architecture. Higher doses (100 and 300 mg kg−1) had comparable effects. In curcumin‐pretreated mice, there was a significant reduction in the degree of both neutrophil infiltration (measured as decrease in myeloperoxidase activity) and lipid peroxidation (measured as decrease in malondialdehyde activity) in the inflamed colon as well as decreased serine protease activity. Curcumin also reduced the levels of nitric oxide (NO) and O2− associated with the favourable expression of Th1 and Th2 cytokines and inducible NO synthase. Consistent with these observations, nuclear factor‐κB activation in colonic mucosa was suppressed in the curcumin‐treated mice. These findings suggest that curcumin or diferuloylmethane, a major component of the food flavour turmeric, exerts beneficial effects in experimental colitis and may, therefore, be useful in the treatment of IBD.

Anti-hyperglycemic effect of black tea (Camellia sinensis) in rat

Investigations were carried out to evaluate the effect of the hot water extract of black tea (Camellia sinensis (L.) O. Kuntze (Theaceae) on streptozotocin (STZ)-induced diabetes in rats. The extract significantly reduced the blood glucose level and was found to possess both preventive and curative effects on experimentally produced diabetes in rats. The study reveals that, like green tea, black tea also possesses antidiabetic activity.

Anti-ulcer effect of the hot water extract of black tea (Camellia sinensis)

The effect of the hot water extract of black tea (Camellia sinensis (L.) O. Kuntze, Theaceae) on ulceration induced by various ulcerogens and by cold restraint stress (CRS) was investigated in albino rats. While prior administration of tea extract for 7 days significantly reduced the incidence of ulcer, ulcer number and ulcer index produced by aspirin, indomethacin, ethanol, reserpine and CRS, it failed to inhibit the ulcers induced by serotonin and histamine. Tea extract also favourably altered the changes in acid and peptic activity of gastric secretion induced by aspirin, indomethacin, ethanol, reserpine and CRS. The observations suggest that the hot water extract of black tea possesses anti-ulcer activity, probably mediated through prostaglandins.

Thearubigin, the major polyphenol of black tea, ameliorates mucosal injury in trinitrobenzene sulfonic acid-induced colitis

Inflammatory bowel disease is characterized by oxidative and nitrosative stress, leukocyte infiltration and upregulation of proinflammatory cytokines. The aim of the present study was to examine the protective effects of thearubigin, an anti-inflammatory and anti-oxidant beverage derivative, on 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis in mice, a model for inflammatory bowel disease. Intestinal lesions (judged by macroscopic and histological score) were associated with neutrophil infiltration (measured as increase in myeloperoxidase activity in the mucosa), increased serine protease activity (may be involved in the degradation of colonic tissue) and high levels of malondialdehyde (an indicator of lipid peroxidation). Both nitric oxide (NO) and O(2)(-) were increased with concomitant upregulation in the mRNA expression of proinflammatory cytokine response and inducible NO synthase (iNOS). Dose-response studies revealed that pretreatment of mice with thearubigin (40 mg kg(-1) day(-1), i.g. for 10 days) significantly ameliorated the appearance of diarrhoea and the disruption of colonic architecture. Higher dose (100 mg kg(-1)) had comparable effects. This was associated with a significant reduction in the degree of both neutrophil infiltration and lipid peroxidation in the inflamed colon as well as decreased serine protease activity. Thearubigin also reduced the levels of NO and O(2)(-) associated with the favourable expression of T-helper 1 cytokines and iNOS. Consistent with these observations, nuclear factor kappa B (NF-kappa B) activation in colonic mucosa was suppressed in thearubigin-treated mice. The results of this study suggest that thearubigin, the most predominant polyphenol of black tea, exerts beneficial effects in experimental colitis and may, therefore, be useful in the treatment of inflammatory bowel disease.

Anti-leukemic activity of Dillenia indica L. fruit extract and quantification of betulinic acid by HPLC

The methanolic extract of Dillenia indica L. fruits showed significant anti-leukemic activity in human leukemic cell lines U937, HL60 and K562. This finding led to fractionation of the methanolic extract, on the basis of polarity, in which the ethyl acetate fraction showed the highest anti-leukemic activity. A major compound, betulinic acid, was isolated from the ethyl acetate fraction by silica gel column chromatography and was identified and characterized. Betulinic acid could explain the anti-leukemic activity of the methanolic extract and the ethyl acetate fraction. Hence the quantitative estimation of betulinic acid was approached in methanolic extract and fractions using HPLC.

Enhancement by oxotremorine of acetylcholine release from the rat phrenic nerve.

1 Oxotremorine (10.5 μM) produced a paralytic effect on twitch responses of rat diaphragm in vitro to direct and indirect stimulation. 2 The paralytic effect of oxotremorine was absent when the diaphragm was stimulated directly in the presence of hemicholinium‐3 (0.42 mM), at a time when twitch responses to indirect stimulation ceased completely. 3 Oxotremorine, at two different pharmacologically active doses, strikingly increased the resting as well as electrically evoked release of acetylcholine into the bathing fluid from the phrenic nerve‐diaphragm preparation. 4 This presynaptic effect of oxotremorine may explain its pharmacological effects at the cholinergic synapses studied so far.

Induction of Apoptosis in Human Leukemic Cell Lines U937, K562 and HL-60 by Litchi chinensis Leaf Extract Via Activation of Mitochondria Mediated Caspase Cascades

The apoptogenic activity of Litchi chinensis leaf extract (LCLE) was investigated against three human leuke- mic cell lines-U937, K562 and HL-60. LCLE inhibited cell growth and metabolic activity of the leukemic cells and showed characteristic features of apoptosis. Flow cytometric analysis showed appreciable number of cells in early and late apoptotic stages. While U937 and K562 cell populations were arrested in the G2-M phase, the HL-60 cell population was arrested in the G1 phase of cell cycle. LCLE induced apoptosis is mediated through mitochondrial intrinsic pathway in- volving the release of cytochrome c into the cytosol and activation of caspase-9 and caspase-3.

Effect of green tea (Camellia sinensis) extract on the rat diaphragm.

The effect of hot water extract of green tea on skeletal muscle and its neurotransmission was studied employing innervated and denervated rat diaphragm. Green tea extract (GTE) has a facilitatory effect at lower concentrations and a paralytic effect at higher concentrations on skeletomotor function. GTE did not have any effect on direct twitch responses or on acetylcholine (ACh) and KCl induced contractures of denervated rat diaphragm and it antagonised the submaximal paralytic effect of D-tubocurarine and decamethonium. GTE-induced facilitation and inhibition were nullified in the presence of magnesium chloride. Nifedipine, reduced GTE-induced facilitation as well as inhibition of twitch responses as a function of its concentration. It was suggested that GTE might act on Ca2+ channels at the skeletomotor junction. The effect of crude polyphenol on neuromuscular junctions was found to be similar to that of GTE. Therefore, it is suggested that the crude polyphenol content of GTE was the active constituent responsible for its effect on neuromuscular junction.

Anti-leukemic activity of Wattakaka volubilis leaf extract against human myeloid leukemia cell lines.

ETHNOPHARMACOLOGICAL RELEVANCE Wattakaka volubilis has been traditionally used in Ayurvedic medicine in India for treatment of several ailments such as bronchial asthma, inflammations, tumors, piles, leucoderma, application to boils, rat bite etc. AIM OF THE STUDY The present study was designed to investigate anti-leukemic activity of the crude aqueous methanolic extract and to identify active compounds from the leaves of Wattakaka volubilis. MATERIALS AND METHODS The leaves of Wattakaka volubilis were extracted with aqueous methanol. Liquid-liquid fractionation of the crude methanolic extract with different organic solvents was done and the fractions were screened for in vitro anti-leukemic activity using different leukemic cell lines. The active fractions were then subjected to chromatographic separation for isolation of bioactive compounds. Structure of isolated compound was elucidated by spectroscopic methods. The in vitro anti-leukemic activities of different extracts of the leaves and isolated compound WVP were studied in U-937, HL-60 and K-562 cell-lines by using cell count, MTT [(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide] and DNA laddering assays, flow-cytometric and confocal microscopic techniques. RESULTS Kaempferol-3-O-[α-l-rhamnopyranosyl-(1→4)-O-α-l-rhamnopyranosyl-(1→6)-O]-β-d-glucopyranoside (WVP) was isolated from crude leaves extract of Wattakaka volubilis. Both the n-butanolic extract (WVB) of Wattakaka volubilis and its isolate WVP were found to be responsible for in vitro anti-leukemic activity. The IC(50) values of WVB were found be 120, 100 and 50(μg/ml) in U937, K562, and HL-60 cell lines, respectively. Whereas, the pure isolate WVP exhibited anti-leukemic activity with IC(50) values of 13.5, 10.8, and 13.2(μg/ml) in U937, K562, and HL-60 cell lines, respectively. The flow-cytometric analysis confirms that the cell cycle arrest occurs at G1 phase in case of U937 and K562 cell lines and G2/M phase in case of HL60 cell lines. Similarly both confocal microsocopic analysis and DNA laddering assay confirm the apoptosis and cell cycle arrests of leukemic cells. CONCLUSION The overall results provide evidence for the ethnopharmacological relevance for use of the plant Wattakaka volubilis in developing novel agents for the treatment of leukemia.

Anti-tremor action of C10Dichol, a peripheral acetylcholine synthesis inhibitor.

1 Anti‐tremor action of decamethylene bis‐(hydroxyethly)‐dimethylammonium bromide (C10Dichol), a peripheral acetylcholine synthesis inhibitor, was investigated. 2 C10Dichol inhibited tremor induced by oxotremorine, nicotine and physostigmine and afforded partial protection from physostigmine‐induced mortality in mice. 3 In non‐paralysing doses, C10Dichol antagonized the neuromuscular effects of oxotremorine, nicotine and physostigmine. 4 Prior administration of C10Dichol failed to prevent tremor and neuromuscular paralysis induced by harmine and arecoline. 5 In the absence of any antimuscarinic property of C10Dichol, its neuromuscular effects appeared to be causually related to its anti‐tremor action. 6 This study reveals a possibility for the development of peripherally acting anti‐Parkinson drugs.