Screening of anti-dengue activity in methanolic extracts of medicinal plants

<p>Abstract</p> <p>Background</p> <p>Dengue fever regardless of its serotypes has been the most prevalent arthropod-borne viral diseases among the world population. The development of a dengue vaccine is complicated by the antibody-dependent enhancement effect. Thus, the development of a plant-based antiviral preparation promises a more potential alternative in combating dengue disease.</p> <p>Methods</p> <p>Present studies investigated the antiviral effects of standardised methanolic extracts of <it>Andrographis paniculata, Citrus limon, Cymbopogon citratus, Momordica charantia, Ocimum sanctum </it>and <it>Pelargonium citrosum </it>on dengue virus serotype 1 (DENV-1).</p> <p>Results</p> <p><it>O. sanctum </it>contained 88.6% of total flavonoids content, an amount that was the highest among all the six plants tested while the least was detected in <it>M. charantia</it>. In this study, the maximum non-toxic dose (MNTD) of the six medicinal plants was determined by testing the methanolic extracts against Vero E6 cells <it>in vitro</it>. Studies also determined that the MNTD of methanolic extract was in the decreasing order of <it>M. charantia </it>><it>C. limon </it>><it>P. citrosum, O. sanctum </it>><it>A. paniculata </it>><it>C. citratus</it>. Antiviral assay based on cytopathic effects (CPE) denoted by degree of inhibition upon treating DENV1-infected Vero E6 cells with MNTD of six medicinal plants showed that <it>A. paniculata </it>has the most antiviral inhibitory effects followed by <it>M. charantia</it>. These results were further verified with an <it>in vitro </it>inhibition assay using MTT, in which 113.0% and 98.0% of cell viability were recorded as opposed to 44.6% in DENV-1 infected cells. Although methanolic extracts of <it>O. sanctum </it>and <it>C. citratus </it>showed slight inhibition effect based on CPE, a significant inhibition was not reflected in MTT assay. Methanolic extracts of <it>C. limon </it>and <it>P. citrosum </it>did not prevent cytopathic effects or cell death from DENV-1.</p> <p>Conclusions</p> <p>The methanol extracts of <it>A. paniculata </it>and <it>M. charantia </it>possess the ability of inhibiting the activity of DENV-1 in <it>in vitro </it>assays. Both of these plants are worth to be further investigated and might be advantageous as an alternative for dengue treatment.</p

Validation of Different Methods of Preparation of Adhatoda vasica Leaf Juice by Quantification of Total Alkaloids and Vasicine

Leaf of Adhatoda vasica (Vasaka) is an important drug of Ayurveda, prescribed as an expectorant. Quinazoline alkaloids present in the leaves are established as active principles. In Ayurveda, its leaf juice (Vasa swarasa) is incorporated in many formulations. Classical method for extracting the juice (swarasa) from the leaf is an elaborate process, which involves subjecting a bolus of crushed fresh leaf to heat followed by squeezing out the juice. Commercially, to prepare the juice of Vasaka, manufacturers have been adopting different methods other than the traditional method. In an effort to evaluate these modified processes phytochemically to identify the process which gives juice of the quality that is obtained by traditional method, in terms of its alkaloid content, we prepared the leaf juice by traditional Ayurvedic method, its modification by steaming of leaf to simulate the traditional method and other methods adopted by some manufacturers. These juice samples were evaluated for the total alkaloid content by spectrophotometric method and vasicine content by thin layer chromatography densitometric method using high performance thin layer chromatography. The high performance thin layer chromatography method was validated for precision, repeatability and accuracy. The total alkaloid content varied from 0.3 mg/ml to 5.93 mg/ml and that of vasicine content varied from 0.2 mg/ml to 5.64 mg/ml in the juice samples prepared by different methods. The present study revealed that steaming of fresh leaves under 15 lb pressure yielded same quantity of juice as the traditional bolus method (25 ml/100 g leaf) and its total alkaloid content and vasicine content (4.05±0.12 and 3.46±0.06 mg/ml, respectively) were very high when compared to the other methods, though the traditional method was found to give the best quality juice with highest amount of total alkaloids (5.93±0.55 mg/ml) and vasicine (5.64±0.10 mg/ml) content

Evaluation of Free Radical Scavenging Activity of an Ayurvedic Formulation, Panchvalkala

We report the free radical scavenging activity of an Ayurvedic preparation Panchvalkala and its individual components (stem bark of Ficus benghalensis, F. glomerata, F. religiosa, F. virens and Thespesia populnea). Being stem barks, these samples contain phenolics (ranging from 3.5% to 10.8% w/w) and tannins (1.6% to 7.0% w/w). This prompted us to study the free radical scavenging activity of Panchvalkala and its components which was evaluated in three in vitro models viz. 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, superoxide radical scavenging activity and reducing power assay. Panchvalkala and its individual components showed significant antiradical activity by bleaching 1,1-diphenyl-2-picrylhydrazyl radical (EC50 ranging from 7.27 to 12.08 µg) which was comparable to pyrogallol (EC50 4.85 µg). Thin layer chromatography of the methanol extracts when sprayed with 0.2% 1,1-diphenyl-2-picrylhydrazyl in methanol revealed several bands with antiradical activity as seen by bleaching of 1,1-diphenyl-2-picrylhydrazyl. All the samples showed good superoxide scavenging potential (EC50 ranging from 41.55 to 73.56 µg) comparable to ascorbic acid (EC50 45.39 µg) in a dose-dependent manner. The reduction ability, Fe3+ to Fe2+ transformation was found to increase with increasing concentrations of all the sample extracts