2Department of Pharmacognosy and Phytochemistry, Gitam University, India
The plants L. repens and A. cymosa were collected from Tirupathi, Chittoor district of Andhra Pradesh was authenticated at Botany Division, Sri Venkateswara University. The voucher specimen (No. 1568 and 1043) were deposited at herbarium and raw drug depository respectively. The plant components had been dried out under shade for Two weeks, coarsely powdered and then kept in air restricted canisters guarded against humidity and sunlight for further study.
About 250g of the powdered crude drug of L. repens and A. cymosa were extracted by cold maceration with 1000 mL of methanol for 18 h after pretreatment with petroleum ether. Both extracts obtained had been concentrated to dryness in the vacuum at 40oC and kept at 4oC inside the refrigerator till further used. The extracts had been subjected to phytochemical and pharmacological evaluation.
The various extract of L. repens and A. cymosa were subjected to qualitative chemical analysis by using standard procedures [11-14].
Adults Wistar albino rats of either sex weighing 180-200g each were used. The animals had been maintained in the regular metallic cages in sets of 6 per cage, with free access to standard diet and drinking water ad libitum in the animal house and kept at room temperature under suitable dietary and environmental circumstances throughout the experiment. The Institutional Animal Ethics Committee examined the complete animal protocols ahead of performing the trials.
Paracetamol, Baker yeast, Distilled water and Digital Thermometer.
To evaluate the degree of toxicity of L. repens methanolic extract, the acute toxicity study was worked based upon OECD (Organization for Economic Cooperation and Development) 423 recommendations to the dose of 2000 mg/Kg. The experimental animals had been noticed for 1h constantly after which hourly for 4h and lastly every 24h up to 14 days for any physical symptoms of the level of toxicity, including writhing, gasping, palpitations and lowered respiratory rate or mortality. No animals died. Therefore the LD50 is greater than 2000 mg/kg. Pre-screening investigation with 200 and 400mg per body weight was done .
The animals were randomly divided into six groups, each group consisting of 10 rats; a total of 60 rats were used in the study by randomized sampling technique: group I (control, normal saline given orally at 2 ml/kg body weight); group II (standard, paracetamol 150 mg/kg); group’s III and IV [methanolic extract of L. repens (MELR)], V and VI [methanolic extract of A. cymosa (MEAC) 200, and 400 mg/kg, respectively]. This is certainly a traditional way of antipyresis screening. Wistar strain of albino rats of either sex weighing 100-200 g was used for the study. The animals had fasted for 18 h before the beginning of the test, yet water was supplied ad libitum. The initial rectal temperature was recorded by using a rectal thermometer to a depth of 1.5 cm in the rectum of rodent. Animals with a body temperature between 36 and 38°C had been as part of the research. A 20% Brewer's yeast in 0.9% w/v saline was injected subcutaneously under the nape of the throat at a dose of 10 ml/kg thereafter. The injection site was rubbed so that the spread of suspension under the skin. Room temperature was maintained at 22-24°C. Following the yeast injection, food was instantly withdrawn. After 10 h post-challenge, the rise in rectal temperature was recorded. Animals which demonstrated an increase in body's temperature to 39°C had been included in the research, enabling the minimum of six rats in every group, the total of 36 rats. The animals received the standard (paracetamol 150 mg/kg) or the test compound (MELR and MEAC 200 and 400 mg/kg) by oral administration, and the rectal temperature was recorded at 0, 30, 60, 90, 120 and 180 min after dosing. The maximum reduction in average rectal temperature in comparison with the control hyperpyrexia group was calculated and compared .
Statistical analysis was carried out using Graph Pad Prism 5.0 (Graph Pad Software, San Diego, CA). All results were expressed as mean ± SD. The data were analyzed by one-way ANOVA followed by Tukey multiple comparison tests.
The MELR and MEAC, when orally administered in the dose of 2000 mg/kg body wt. did not produce any significant changes in the autonomic or behavioural responses, including death during the observation period.
The phytochemical screening for methanolic extracts of L. repens and A. cymosa were carried out, and results were displayed in Table 1.
|Zn. Hydrochloride test||+||+|
|Lead acetate Test||+||+|
|Volatile oil||Stain test||-||-|
|Tannins & Phenols||Fecl3 Test||+||+|
|Potassium dichromate test||+||+|
|Acid compounds||Litmus test||+||-|
|Amino acids||Ninhydrin test||+||+|
“+” – Present and “-” - Absent
Subcutaneous injection of the pyrogenic dose of yeast produced elevated changes in rectal temperature, which is shown in Table 2. The MELR and MEAC showed a significant (P<0.05) decrease in rectal temperature at dose 400 mg/kg when compared with the standard. Extract at doses of 200, and 400 mg/kg showed a progressive decline in mean temperature pattern with the increase in the dose. Paracetamol showed significant (P<0.001) decrease in rectal temperature. The onset of action of paracetamol was 30 min. The line graph (Fig. 1) shows that paracetamol registered a phenomenal decrease in mean temperature from 38.7±0.12 to 37.01±0.42. Figure 2 illustrates that the most significant (P < 0.05) decrease in round-the-clock mean temperature in this study was shown by paracetamol followed by MELR and MEAC at 400 mg/kg and 200 mg/kg. Statistical analysis was performed using one-way analysis of variance followed by Tukey's test. The consequences of extract at a dose of 400 mg/kg were nearly much like that of standard drug paracetamol. The results were recorded as mean ± SD. Out of all the groups apart from negative control, the temperature became normal through 3 h of study (Figure 1, Table2).
|Groups||BBT||Rectal Temperature (°C) after 10h of yeast injection|
|0 min||30 min||60 min||90 min||120 min||150 min||180 min|
|Control||37.12 ± 0.23||38.68 ± 0.21||38.65 ± 0.25||38.91 ± 0.35||38.83 ± 0.32||38.81 ± 0.18||38.73 ± 0.58||38.45 ± 0.43|
|Standard (Paracetamol 150 mg/kg)||37.16 ± 0.21||38.7 ± 0.12#||37.96 ± 0.18#||37.3 ± 0.45#||36.85 ± 0.15#||36.92 ± 0.43#||36.96 ± 0.22#||37.01 ± 0.42#|
|MELR (200 mg/kg)||37.25 ± 0.19||38.78 ± 0.33a||38.75 ± 0.37a||38.63 ± 0.13a||38.66 ± 0.12a||38.55 ± 0.3a||38.51 ± 0.23a||38.43 ± 0.16a|
|MELR (400 mg/kg)||37.18 ± 0.22||38.65 ± 0.42b||38.55 ± 0.23b||38.43 ± 0.39b||38.18 ± 0.35b||37.95 ± 0.54b||37.85 ± 0.44b||37.65 ± 0.23b|
|MEAC (200 mg/kg)||37.08 ± 0.16||38.73 ± 0.26||38.83 ± 0.46||38.75 ± 0.23||38.36 ± 0.37||38.31 ± 0.36||38.11 ± 0.58||38. 08 ± 0.52|
|MEAC (400 mg/kg)||37.16 ± 0.13||38.95 ± 0.18b||38.53 ± 0.33b||38.26 ± 0.33b||37.85 ± 0.53b||37.5 ± 0.38b||37.35 ± 0.43b||37.13 ± 0.48b|
- Metrouh-Amir H., et al. "Evaluation in vivo of anti-inflammatory and analgesic properties of Matricaria pubescens alkaloids". South African Journal of Botany 116 (2018): 168-174.
- Husseini Y., et al. "Analgesic and anti-inflammatory activities of hydro-alcoholic extract of Lavandula officinalis in mice: possible involvement of the cyclooxygenase type 1 and 2 enzymes". Revista Brasileira de Farmacognosia 26.1 (2016): 102-108.
- Les DH. "Aquatic dicotyledons of North America: ecology, life history, and systematics: CRC Press". (2017).
- Hsu H., et al. "Oriental materia medica: a concise guide: Keats Publishing (1996).
- Pullaiah T. "Encyclopaedia of world medicinal plants: Daya books" (2006).
- Brahmachari G. "Limnophila (Scrophulariaceae): Chemical and Pharmaceutical Aspects-An Update". The Open Natural Products Journal 7.4 (2014).
- Do QD., et al. "Effect of extraction solvent on total phenol content, total flavonoid content, and antioxidant activity of Limnophila aromatica". Journal of food and drug analysis 22 (204): 296-302.
- Kukongviriyapan U., et al. "Antioxidant and vascular protective activities of Cratoxylum formosum, Syzygium gratum and Limnophila aromatica". Biological and Pharmaceutical Bulletin 30(2007): 661-666.
- Rao JV., et al. "Antimicrobial activity of the essential oil of Limnophila gratissima". Fitoterapia 60.4 (1989): 376-377.
- Nanasombat S., et al. "Antimicrobial, antioxidant and anticancer activities of Thai local vegetables". Journal of Medicinal Plants Research 3.5(2009): 443-449.
- Suksamrarn A., et al. "Antimycobacterial and antioxidant flavones from Limnophila geoffrayi". Archives of pharmacal research 26.10 (2003): 816-820.
- Marles RJ., et al. "Antidiabetic plants and their active constituents". Phytomedicine 2.2 (1995): 137-189.
- Galani VJ., et al. "Psychotropic activity of Argyreia speciosa roots in experimental animals". Ayu 32.2 (2011): 380.
- Packialakshmi N., et al. "Antibacterial Screening on Leaves of Argyreia Cymosa Roxb. Against Pathogenic Bacteria Isolated from Infected Pateints Samples Wound, Sputum and Stool". International Journal of Applied Sciences and Biotechnology 2(3): 279-282.
- Kiran PM., et al. "Investigation of hepatoprotective activity of Cyathea gigantea (Wall. ex. Hook.) leaves against paracetamol-induced hepatotoxicity in rats". Asian Pacific journal of tropical biomedicine 2.5 (2012): 352.
- Vogel HG. "Drug discovery and evaluation: pharmacological assays: Springer Science & Business Media". (2002).
- Bhattacharya A., et al. "Antipyretic effect of ethanolic extract of Moringa oleifera leaves on albino rats". Tanta Medical Journal 42.2 (2014): 74.
- Zeisberger E. From humoral fever to neuroimmunological control of fever". Journal of Thermal Biology 24.5 (1999): 287-326.
- Banks WA., et al. "Permeability of the blood-brain barrier to soluble cytokine receptors". Neuroimmunomodulation 2.3(1995):161-165.
- Roth J., et al. "Signaling the brain in systemic inflammation: role of sensory circumventricular organs". Frontiers in Bioscience 9.7(2004): 290-300.
- Schiltz JC., et al. "Signaling the brain in systemic inflammation: the role of perivascular cells". Frontiers in Bioscience 8 (2003): 1321-1329.
- Saper CB., et al. "The neurologic basis of fever". The New England Journal of Medicine 330.26 (1994): 1880-1886.
- Gupta M., et al. "Anti-inflammatory and antipyretic activities of β-sitosterol". Planta medica 39.6 (1980): 157-163.
- Backhouse N., et al. "Anti-inflammatory and antipyretic effects of boldine". Agents and actions 42.3 (1994): 114-117.
- Chang C-P., et al. "flavonoid baicalin protects against cerebrovascular dysfunction and brain inflammation in experimental heatstroke". Neuropharmacology 52.2(2007): 1024-1033.
- Li S., et al. "Acetaminophen: antipyretic or hypothermic in mice? In either case, PGHS-1b (COX-3) is irrelevant". Prostaglandins Other Lipid Mediat. 85.5 (2008): 89-99.