STANDARDIZATION OF VELLARUGU CHOORANAM: A SIDDHA HERBAL DRUG

Pharmacognostical standardization of herbal formulation is essential in order to assess the quality of drugs, based on the concentration of their active principles. Vellarugu chooranam is a reputed drug mentioned in the ancient books of Siddha Medicine for the treatment of Vata diseases, arthritis, constipation, and diabetes mellitus. The main component of Vellarugu chooranam is whole plant of Enicostemma littorale Blume (Siddha Name: Vellarugu) (Gentianaceae). In the present study an attempt has been made to standardize the Vellarugu chooranam. For the standardization of this drug Organoleptic properties, Phytochemical screening, Fluorescence analysis, Elemental analysis, Physicochemical parameters such as moisture content, ash values, extractability in water and ethanol were carried out. TLC and HPLC fingerprints of the Vellarugu chooranam were also prepared to evaluate its quality. These set of parameters were found to be sufficient to evaluate authenticity of the Vellarugu chooranam and can be used as reference standards for the preparation of a standardized pharmaceutical product and further quality control researches

Evaluation of the toxic potential of standardized extracts (hot water extract and cold ethanolic extract) of Trichosanthes cucumerina Linn. aerial parts

Trichosanthes cucumerina Linn. Es una de las plantas comunmente utilizadas en el sistema de medicina tradicional de Sri Lanka, en la preparación de formulaciones para el tratamiento de diversas enfermedades. Debido a que los efectos tóxicos de T. cucumerina no se conocen, los objetivos de este estudio fueron: (a) estandarizar los extractos obtenidos con agua caliente (EAC) y con etanol frío (EEF), y (b) evaluar la toxicidad de ambos extractos. Ambos extractos fueron estandarizados por obtención de sus densitogramas y huella digital con HPLC. Adicionalmente se determinaron parámetros fisicoquímicos, tales como: cenizas totales, cenizas solubles en agua y cenizas solubles en ácido. La administración de EAC y EEF a ratones no mostró efectos tóxicos agudos ni crónicos. Las funciones renales, hepáticas, estudios hematológicos y de fertilidad en machos y hembras fueron normales. Se concluye que los extractos estandarizados de T. cucumerina, a las dosis ensayadas no producen ningún efecto tóxico.Trichosanthes cucumerina Linn. is one of the medicinal plants that is often used in Sri Lankan traditional systems of medicine for the preparation of formulations to treat a variety of disease conditions. However, the toxic effects of T. cucumerina are not known. The aims of the present study were to (a) standardize hot water (HWE) and cold ethanolic (CEE) extracts of T. cucumerina aerial parts, and (b) evaluate toxic potential of the plant extracts. Both extracts were standardized by developing their densitograms and HPLC fingerprints and determination of physico – chemical parameters such as total ash, water soluble ash and acid insoluble ash. Administration of the HWE or CEE to mice did not result in acute or chronic toxic effects as evident from their effects on (a) liver and kidney functions and (c) hematological parameters and (d) fertility of male or female mice.In conclusion, the results of this study have revealed that standardized extracts of T. cucumerina at the doses tested do not produce any serious toxic side effects

Protection against diethylnitrosoamine-induced hepatocarcinogenesis by an indigenous medicine comprised of Nigella sativa, Hemidesmus indicus and Smilax glabra: a preliminary study

BACKGROUND: A decoction comprised of Nigella sativa seeds, Hemidesmus indicus root and Smilax glabra rhizome is used to treat cancer patients in Sri Lanka. However, the anti-carcinogenic properties of this decoction have not been experimentally confirmed. The purpose of this study was to determine whether the above decoction could protect against chemically induce hepatocarcinogenesis. METHODS: The effects of this decoction on diethylnitrosamine (DEN) induced hepatocarcinogenesis were examined in male Wistar rats using the medium term bioassay system of Ito, based on a 2-step model of hepatocarcinogenesis. Rats were randomly divided into 6 groups of 10 each. Groups 1 to 4 were injected with DEN (200 mg/kg) to initiate carcinogenesis. Twenty-four hours later groups 1 and 2 were administered the decoction at 4 g/kg body weight/day (dose 1) and 6 g/kg body weight/day (dose 2), respectively. Group 3 and group 4 were given distilled water instead of the decoction and a suspension of garlic powder (20 g/kg body weight/day) in distilled water (positive control), respectively. Group 5 and 6 were injected with normal saline and twenty-four hours later group 5 was given distilled water (normal control) while group 6 was given decoction dose 2 (decoction control). Oral feeding continued for two weeks after which all rats were subjected to 2/3 partial hepatectomy to promote carcinogenesis. Oral feeding continued for eight more weeks. At the end of the 10th week, rats were sacrificed and samples of livers taken for immunohistochemical studies. Carcinogenic potential was scored by comparing the number, area and staining intensity of glutathione S-transferase placental form (GST-P) positive foci and the number of cells/cm(2 )of the positive foci in the livers of the six groups of rats. RESULTS: The number and area of DEN-mediated GST-P positive foci, number of cells/cm(2 )of foci and staining intensity of the foci were significantly (P > 0.001) reduced by the decoction and garlic in the order dose 2 = garlic >dose 1. CONCLUSION: Overall results indicate that the decoction comprised of N. sativa, S. glabra and H. indicus has the potential to protect rat liver against DEN induced hepatocarcinogenesi

In vitro Cytotoxic and Antioxidant Activity of Leaf Extracts of Mangrove Plant, Phoenix paludosa Roxb

Purpose: To investigate the anti-proliferative and antioxidant potentials of four different solvent extracts of Phoenix paludosa Roxb leaves.Methods: Four different solvent (hexane, chloroform, ethyl acetate and methanol) leaf extracts of the plant were tested for cytotoxicity against four cancer cells, viz, MCF-7 (oestrogen positive breast cancer cell line), MDA-MB-231 (triple negative breast cancer cell line), SK-BR-3 (breast adenocarcinoma) and ACHN (renal adenocarcinoma) as well as two normal cell lines, namely, HEK-293 (embryonic kidney cells) and MCF-10A (normal mammary epithelial cells)]. 2, 2-Diphenyl-1-picrylhydrazyl (DPPH) and 2, 29-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) free radical scavenging assays were used to evaluate the antioxidant activity of the crude extracts.Results: The methanol extract showed the highest antioxidant activity (DPPH, half maximal inhibitory concentration (IC50) = 30.17 ± 6.21 μg/mL) and (ABTS, IC50 = 27.91 ± 3.21 μg/mL). Of the four extracts, methanol extract showed the strongest significant (p < 0.05) cytotoxicity to all four cancer cell lines at 24 and 48 h of incubation followed by the chloroform extract (IC50 of methanol extract (24 and 48 h): 36.71 ± 8.72 and 33.19 ± 5.53 μg/mL (MCF-7), 159.7 ± 32.09 and 141.9 ± 26.2 μg/mL (MDA-MB-231), 103.3 ± 18.9 and 75.39 ± 19.39 μg/mL (SKBR-3), 57.21 ± 3.72 and 43.16 ± 10.25 μg/mL (MCF-10A), 37.48 ± 5.75 and 26.99 ± 1.85 (ACHN) and 66.83 ± 14.26 and 60.34 ± 10.66 μg/mL (HEK-293)). Furthermore, the methanol extract was least cytotoxic to normal cell lines.Conclusion: The results obtained indicate that the methanol leaf extract of P. paludosa exhibit potent antioxidant and cytotoxic activities and has the potential of being developed into an anti-cancer agent.Keywords: Phoenix paludosa, antiproliferative, antioxidant, cytotoxicit

Anti-Inflammatory Activity Is a Possible Mechanism by Which the Polyherbal Formulation Comprised of Nigella sativa

The present study investigated the anti-inflammatory effects of a polyherbal decoction comprised of Nigella sativa, Hemidesmus indicus, and Smilax glabra in order to justify its claimed antihepatocarcinogenic activity. Activation of hepatic nuclear factor-kappa B (NF-κB), IκB kinase (IKK α/β) proteins, and TNFα and IL-6 expression was investigated in diethylnitrosamine- (DEN-) induced C3H mice-bearing early hepatocarcinogenic changes. Acute phase inflammatory response was evaluated by carrageenan-induced rat paw edema formation. Anti-inflammatory mechanisms were also assessed by determining effect on (a) membrane stabilization, (b) nitric oxide (NO) inhibitory activity, and (c) inhibition of leukocyte migration. A significant inhibition of the paw edema formation was observed in healthy rats as well as in rats bearing early hepatocarcinogenic changes with oral administration of the decoction. As with the positive control, indomethacin (10 mg/kg b.w.) the inhibitory effect was pronounced at 3rd and 4th h after carrageenan injection. A notable IKK α/β mediated hepatic NF-κB inactivation was associated with a significant hepatic TNFα downregulation among mice-bearing hepatocarcinogenic changes subjected to decoction treatment. Inhibition of NO production, leukocyte migration, and membrane stabilization are possible mechanisms by which anti-inflammatory effect is mediated by the decoction. Overall findings imply that anti-inflammatory activity could be one of the mechanisms by which the decoction mediates its antihepatocarcinogenic effects

Modulation of apoptosis in human hepatocellular carcinoma (HepG2 cells) by a standardized herbal decoction of Nigella sativa seeds, Hemidesmus indicus roots and Smilax glabra rhizomes with anti- hepatocarcinogenic effects

<p>Abstract</p> <p>Background</p> <p>A standardized poly-herbal decoction of <it>Nigella sativa </it>seeds, <it>Hemidesmus indicus </it>roots and <it>Smilax glabra </it>rhizomes used traditionally in Sri Lanka for cancer therapy has been demonstrated previously, to have anti-hepatocarcinogenic potential. Cytotoxicity, antioxidant activity, anti-inflammatory activity, and up regulation of p53 and p21 activities are considered to be some of the possible mechanisms through which the above decoction may mediate its anti-hepatocarcinogenic action. The main aim of the present study was to determine whether apoptosis is also a major mechanism by which the decoction mediates its anti-hepatocarcinogenic action.</p> <p>Methods</p> <p>Evaluation of apoptosis in HepG2 cells was carried out by (a) microscopic observations of cell morphology, (b) DNA fragmentation analysis, (c) activities of caspase 3 and 9, as well as by (d) analysis of the expression of pro-apoptotic (Bax) and anti-apoptotic (Bcl-2) proteins associated with cell death.</p> <p>Results</p> <p>The results demonstrated that in HepG2 cells, the decoction can induce (a) DNA fragmentation and (b) characteristic morphological changes associated with apoptosis (nuclear condensation, membrane blebbing, nuclear fragmentation and apoptotic bodies). The decoction could also, in a time and dose dependent manner, up regulate the expression of the pro-apoptotic gene <it>Bax </it>and down regulate expression of anti-apoptotic <it>Bcl-2 </it>gene (as evident from RT-PCR analysis, immunohistochemistry and western blotting). Further, the decoction significantly (<it>p </it>< .001) enhanced the activities of caspase-3 and caspase-9 in a time and dose dependent manner.</p> <p>Conclusions</p> <p>Overall findings provide confirmatory evidence to demonstrate that the decoction may mediate its reported anti-hepatocarcinogenic effect, at least in part, through modulation of apoptosis.</p

In vitro and in vivo evaluation of antioxidant activity of Trichosanthes cucumerina aerial parts

The present study was conducted to determine whether aerial parts of Trichosanthes cucumerina extracts can exert significant antioxidant activity. The antioxidant activity of a hot water extract (HWE) and a cold ethanolic extract (CEE) of T. cucumerina aerial parts was evaluated by assessing its (a) radical scavenging ability and prevention effect of lipid peroxidation in vitro, and (b) effects on lipid peroxidation and antioxidant enzyme activities, in vivo. In vitro antioxidant assays (DPPH, TBARS and carotene-linoleic acid assays) clearly demonstrated the antioxidant potential of HWE and CEE. Moreover, HWE increased SOD: by 91.2% and GPX by 104.4% while CEE increased SOD: by 115.5% and GPX by 96.4%) in CCl4-induced rats. Treatments with HWE and CEE prevented the accumulation of lipid peroxidation products by 30.5% and 33.8%, respectively, in liver tissues compared to the rats exposed only to CCl4. In conclusion, the present investigation demonstrates for the first time that components in T. cucumerina aerial parts can exert significant antioxidant activity in vivo and in vitro

Effect of Standardized Decoction of Nigella sativa Seed, Hemidesmus indicus Root and Smilax glabra Rhizome on the Expression of p53 and p21 Genes in Human Hepatoma Cells (HepG2) and Mouse Liver with Chemically-Induced Hepatocarcinogenesis

Purpose: To evaluate in vitro (using human hepatoma HepG2 cells) and in vivo (using mouse liver with diethlynitrosamine (DEN)-induced hepatocarcinogenesis) effect of a standardized decoction on the expression of p53 (tumour suppressor) and p21 (cyclin kinase inhibitor) genes with the long-term goal of developing the formulation into a globally acceptable therapy for hepatocellular carcinoma (HCC). Methods: The effect of the decoction on (a) mRNA and (b) protein expression of p53 and p21 genes in HepG2 cells and mouse livers with DEN-induced early hepatocarcinogenesis were evaluated by (a) reverse transcription PCR (RT-PCR) and (b) immunohistochemical and Western blot analysis, respectively. Results: The results demonstrated that the decoction significantly (p <0.001) enhanced the expression of p53 and p21 genes in a time- and dose-dependent manner in HepG2 cells. A dose of 75 μg/ml significantly increased p53 mRNA at 24 and 48 h and p21 mRNA at 12, 24, 48 h of incubation with the decoction (p <0.01). Induction of hepatocarcinogenesis in mice significantly increased hepatic expression of both p53 and p21 compared to distilled water control (p <0.001), while treatment with the decoction further enhanced expression of both genes in DEN-induced hepatocarcinogenesis (p <0.01). Conclusion: Overall, the findings demonstrate that the decoction may mediate its reported antihepatocarcinogenic effect, at least in part, through the modulating activities of genes involved in tumour suppression and cell cycle arrest

The gastroprotective effect of ethyl acetate fraction of hot water extract of Trichosanthes cucumerina Linn and its underlying mechanisms

Abstract Background Antacids, anticholinergic drugs, histamine H2- receptor antagonists and irreversible proton pump inhibitors have been used for the treatment of gastric ulcers. However, prolonged use of these drugs may lead to series of adverse effects such as diarrhea, headache, rash, hypertension, muscular and joint pain. Therefore, there is an urgent need of more effective and safer treatments with fewer side effects. The aim of the present study was to scientifically evaluate the gastroprotective activity of fractions of the hot water extract of Trichosanthes cucumerina Linn (Family: Cucurbitaceae) aerial parts with a view to identifying the fraction with the best gastroprotective activity and the possible mechanism/s by which this fraction exert gastroprotection. Methods Gastroprotective activity of hexane fraction (HF), ethyl acetate fraction (EF), butanol fraction (BF) and aqueous fraction (AF) were evaluated by the assessment of ability to reduce the ulcer index in ethanol-induced rat model and the mode of action by which the most active fraction mediating gastroprotection. Results EF showed the maximum gastroprotection effect followed by BF and AF. EF (75 mg/kg) exhibited significantly higher gastroprotection compared to the reference drugs. Further investigations with two lower doses of EF confirmed that EF can mediated a significant and dose dependent gastroprotection. The rats treated with the EF showed significant reduction in free acidity (45%), total acidity (by 48%) in the gastric juice, increased the amount of mucus produced by the rat gastro mucosa and potent antihistamine activity (by 25.6%). EF was also rich in phenolic compounds and flavonoids. Conclusion Gastroprotective mechanism of EF is possibly involves inhibition of acidity, elevation in mucus content, inhibition of histamine and antioxidant mechanisms