Rajani Mathur

Cytotoxic and antitumor effects of brucine on Ehrlich ascites tumor and human cancer cell line

AIMS Brucine (BRU), a natural plant alkaloid is reported to possess cytotoxic and antiproliferative activities. In this study we aimed to investigate its in vitro and in vivo antitumor and antiangiogenic effects. MAIN METHODS Cell proliferation and viability was assessed using microculture tetrazolium tests (MTT). As predictive markers we determined intracellular levels of vascular endothelial growth factor (VEGF), Interleukin-12 (IL-12), tumor necrosis factor (TNF-α), caspase-3, -8 and -9 by ELISA and enzymatic activity assays. In addition, anti-VEGF neutralization effect was evaluated to assess whether it could result in augmented anticancer efficacy than the single agent. Antitumor activity was evaluated against Ehrlich ascites and solid tumor models. 15×10(6) EAC cells were implanted intraperitoneally (i.p., ascites tumor) and subcutaneous (s.c., solid tumor) in Swiss albino mice. Mice with established tumors received brucine i.p. at 12.5, 25, and 50mg/kg for 14days in ascites tumor and 50mg/kg in solid tumor for 30days. Tumor volume, cell viability, angiogenic, anti-angiogenic, anti-inflammatory factors and antioxidant parameters were determined. Immunohistochemistry analysis for VEGF and CD-31 was also performed. KEY FINDINGS BRU produced time and dose-dependent inhibition of MCF-7 in vitro and EAC tumors in vivo. The anti-angiogenic effects were accompanied with decreased VEGF and TNF-α and increased IL-12 expression. BRU reduced peritoneal angiogenesis and microvessel density in vivo. CONCLUSION Our findings suggest that BRU possesses antitumor and anti-angiogenic activities in vitro and in vivo. The above results showed that BRU can be used as a potential anticancer agent as an antimetastatic and anti-angiogenic agent.

Antitumor properties of Boswellic acid against Ehrlich ascites cells bearing mouse.

Boswellic acid (BA), a triterpene, isolated from Boswellia serrata (Burseraceae) has been found to possess potent anti-inflammatory and anti-cancer activity. The present study aimed at exploring the possible role of BA on ascites and solid Ehrlich tumor. Ascitic tumor development was evaluated 14 d after tumor implantation by quantification of the ascitic fluid volume whereas solid tumor was evaluated after 30 d tumor implantation by H&E and IHC. The i.p. administration of BA significantly inhibited ascitic and solid Ehrlich tumor model. This inhibition was observed with reduced ascitic volume, solid tumor volume and body weight when compared to control mice. The treatments also increased the survival of tumor-bearing mice. VEGF and TNF- α levels were decreased, whereas the IL-12 levels were increased with BA treatment at 25mg/kg. Further, results on decrease in the peritoneal angiogenesis and microvessel density showed the anti-angiogenic potential. Microscopic examination of tumors revealed that in BA-treated groups the expression of Bax and caspase 3 increased, suggesting drug induced tumor cell apoptosis through activating the pro-apoptotic bcl-2 family and caspase-3. The present study sheds light on the potent antitumor property of the boswellic acid and can be extended further to develop therapeutic protocols for treatment of cancer.

Boswellic acid inhibits inflammatory angiogenesis in a murine sponge model

The aim of the present study was to investigate the effects of boswellic acid (BA) on key components of inflammatory angiogenesis in the murine cannulated sponge implant angiogenesis model. Polyester-polyurethane sponges, used as a framework for fibrovascular tissue growth, were implanted in Swiss albino mice and BA (12.5 or 25mg/kg/day) was given through installed cannulas for nine days. The implants collected at day 9 post-implantation were processed for the assessment of hemoglobin (Hb). Relevant levels of inflammatory, angiogenic and fibrogenic cytokines were also determined. BA treatment resulted in significant decrease in sponge vascularization (Hb content) and in vascular endothelial growth factor (VEGF) and transforming growth factor (TGF-β1) at both doses. Further, BA decreased expression of VEGF and CD31 and reduced % microvessel density (MVD) in sponge implants. A regulatory function of BA on multiple parameters of the main components of inflammatory angiogenesis has been revealed giving an insight into the potential therapeutic use underlying the actions of BA.

Evaluation of Wound Healing Potential of Topical Formulation of Leaf Juice of Tridax Procumbens L. in Mice

Tridax procumbens L. (Compositae) is a common weed that grows in the rice fields of India. Traditionally the juice from leaves of Tridax procumbens has been used for healing dermal wound. However, in experimental studies, equivocal pro and anti-healing action of T. procumbens has been demonstrated. The present study evaluates the effect of topical ointment formulation of the leaf juice of T. procumbens using excision wound model in mice. Excision wounds (4 mm, i.d.) were inflicted on depilated back of mice. Ointment formulation of TP (50 mg of either 1 or 4 mg/g) was applied twice daily for 4 days on the dermal wound. Similarly, control group was treated with VEGF ointment (50 mg of 1 μg/g). The parameters observed were re-epithelization, vascularity, fibroblast number, collagen content. The healing exerted by TP (1 mg/g) was comparable to VEGF (1 μg/g). On the other hand, TP (4 mg/g) induced inflammation, edematous tissue and decreased vascularity. Taken together, the results imply that TP possesses dose dependent pro-healing potential, and its high dose exerts inflammatory reaction.

Psidium guajava Linn. leaf extract affects hepatic glucose transporter-2 to attenuate early onset of insulin resistance consequent to high fructose intake: An experimental study.

Background: Insulin resistance (IR) is amalgam of pathologies like altered glucos metabolism, dyslipidemia, impaired glucose tolerance, non-alcoholic fatty liver disease, and associated with type-II diabetes and cardiometabolic diseases. One of the reasons leading to its increased and early incidence is understood to be a high intake of processed fructose containing foods and beverages by individuals, especially, during critical developmental years. Objective: To investigate the preventive potential of aqueous extract of Psidium guajava leaves (PG) against metabolic pathologies, vis-à-vis, IR, dyslipidemia, hyperleptinemia and hypertension, due to excess fructose intake initiated during developmental years. Materials and Methods: Post-weaning (4 weeks old) male rats were provided fructose (15%) as drinking solution, ad libitum, for 8 weeks and assessed for food and water/fructose intake, body weight, fasting blood sugar, mean arterial pressure, lipid biochemistry, endocrinal (insulin, leptin), histopathological (fatty liver) and immunohistochemical (hepatic glucose transporter [GLUT2]) parameters. Parallel treatment groups were administered PG in doses of 250 and 500 mg/kg/d, po × 8 weeks and assessed for same parameters. Using extensive liquid chromatography-mass spectrometry protocols, PG was analyzed for the presence of phytoconstituents like Myrecetin, Luteolin, Kaempferol and Guavanoic acid and validated to contain Quercetin up to 9.9%w/w. Results: High fructose intake raised circulating levels of insulin and leptin and hepatic GLUT2 expression to promote IR, dyslipidemia, and hypertension that were favorably re-set with PG. Although PG is known for its beneficial role in diabetes mellitus, for the first time we report its potential in the management of lifelong pathologies arising from high fructose intake initiated during developmental years.

Qualitative and quantitative assessment of four marketed formulations of Brahmi

This study was conducted with the aim to compare two batches each of four popular commercial formulations of Bacopa monnieri (Brahmi), and report, if any, inter-batch variations. The formulations were procured from local market and analyzed for label specifications, uniformity of weight of capsule, identity, purity and strength parameters (total ash content test, acid insoluble ash content, water soluble extractive, alcohol soluble extractive, loss on drying). Bacoside A, one of the pharmacologically active saponin present in B. monnieri, was quantified in all the formulations using UV-spectrophotometer. In addition each formulation was assessed and compared for variation in biological activity using in vitro test for hemolytic activity using human erythrocytes. The results of the study show that there is a wide variation in the quality and content of herbal drugs marketed by different manufacturers. More importantly this study demonstrates that there exists a bigger challenge of batch-to-batch variation in the quality and content of herbal formulations of the same manufacturer. This challenge of providing standardized formulations is being faced by not any one manufacturing house but by all, and may be attributed firstly to, lack of stringent regulations and secondly to high variability in raw material quality.

Investigation of anti-asthmatic potential of Kanakasava in ovalbumin-induced bronchial asthma and airway inflammation in rats

ETHNO-PHARMACOLOGICAL RELEVANCE Kanakasava is an Indian traditional Ayurvedic formulation containing Datura (Datura metel), Vasaca (Adhatoda vasica), Dhataki (Woodfordia fruticosa) and Grape (Vitis vinifera) extracts as major constituents and used to treat pulmonary diseases including coughing, breathing difficulty and asthma. The present study was designed to assess the safety and therapeutic efficacy of Kanakasava against ovalbumin-induced bronchial asthma and related airway inflammation in rats due to lack of evidence based therapeutic efficacy data. MATERIAL AND METHODS Male wistar rats were sensitized with allergen (ovalbumin, 40mg/rat+aluminum hydroxide, 2.0mg/rat) and treated orally with standard dexamethasone (2.5mg/kg, b.w.) or Kanakasava (1.23 and 2.46ml/kg, b.w.) from day 1 to day 28. Inflammatory markers, including cell counts and cytokines such as interleukins (IL-4, IL-5, IL-1β), tumor necrosis factor (TNF-α), leukotriene (LTD-4), immunoglobulin (IgE), nitric oxide and nitrite levels in both blood and broncheo alveolar lavaged fluid (BALF) were analyzed. Abdominal mesentery was studied histologically for mast cell degranulation, whereas lung functions were investigated by spirometer. Method was also developed to quantify gallic acid and ethyl gallate content in Kanakasava by HPTLC for its quality control. RESULTS None of the rats exhibited mortality and Kanakasava was found to be safe at the tested doses. Treatment with Kanakasava significantly (P<0.01) reversed elevated levels of IgE, cytokines, nitrites and influx of eosinophils and neutrophils in blood and BALF. These findings were further supported by the significant improvement in lung functions (P<0.01) and suppression (P<0.01) of degranulation of mast cells. The content of gallic acid and ethyl gallate in Kanakasava was found to be 1.94% and 0.98%, respectively. CONCLUSION These findings demonstrated the preventive effect of Kanakasava in allergen induced model of asthma providing scientific basis for its traditional use in Ayurveda, since long time.

Interaction of aqueous leaf extract of Aegle marmelos (L.) Corr. with cholinergic, serotonergic and adrenergic receptors: An ex vivo study

Objectives: The aim was to study interaction of aqueous leaf extract of Aegle marmelos (AM) with cholinergic, serotonergic, and adrenergic receptor systems using appropriate rat tissues-ileum, fundus and tracheal chain, respectively. Materials and Methods: Cumulative concentration-response curves (CRC) were constructed at various doses on each tissue for AM and respective standard agonist. The CRC was again plotted in presence and absence of respective standard antagonist to confirm the interaction of receptor system and AM. Results: AM induced concentration-dependent contractions in isolated rat ileum (0.2–6.4 mg/ml) and fundus (0.2–3.2 mg/ml) that were inhibited significantly (P < 0.05) in the presence of atropine (10−7 M) and ketanserin (10−6 M), respectively. The relaxant effect, produced by AM (0.2 mg/ml) on carbachol (10−5 M) precontracted rat tracheal chain, was also inhibited significantly (P < 0.05) by propranolol (1 ng/ml). Conclusion: It may be concluded that AM possesses agonistic activity on cholinergic, serotonergic and adrenergic receptors.

Development and validation of high-performance thin-layer chromatography method for simultaneous determination of polyphenolic compounds in medicinal plants

Context: Quantitative standardization of plant-based products is challenging albeit essential to maintain their quality. Aims: This study aims to develop and validate high-performance thin-layer chromatography (HPTLC) method for the simultaneous determination of rutin (Ru), quercetin (Qu), and gallic acid (Ga) from Psidium guajava Linn. (PG) and Aegle marmelos (L.) Correa. (AM) and correlate with antioxidant activity. Materials and Methods: The stock solution (1 mg/mL) of standard Ru, Qu, and Ga in methanol: Water (1:1) was serially diluted and spotted (5 μL) on slica gel 60 F254thin-layer chromatography plates. Toluene: Ethyl acetate: Formic acid: Methanol (3:4:0.8:0.7, v/v/v) was selected as mobile phase for analysis at 254 nm. Hydroalcoholic (1:1) extracts of leaves of PG and AM were fractionated and similarly analyzed. Antioxidant activity was also determined using 2, 2-diphenyl-1-picrylhydrazyl assay. Results: The developed method was robust and resolved Ru, Qu, and Ga at Rf0.08 ± 0.02, 0.76 ± 0.01, and 0.63 ± 0.02, respectively. The intra-day, interday precision, and interanalyst were <2% relative standard deviation. The limit of detection and limit of quantification for Ru, Qu, and Ga were 4.51, 4.2, 5.27, and 13.67, 12.73, 15.98 ng/spot, respectively. Antioxidant activity (Log 50% inhibition) of PG and AM was 4.947 ± 0.322 and 6.498 ± 0.295, respectively. Conclusion: The developed HPTLC method was rapid, accurate, precise, reproducible, and specific for the simultaneous estimation of Ru, Qu, and Ga.

Aegle marmelos impedes onset of insulin resistance syndrome in rats provided with drinking fructose from weaning to adulthood stages of development — a mechanistic study

In this study, we explored the effect of aqueous extract of leaves of Aegle marmelos (AM) on hepatic carbohydrate metabolism and insulin downstream signalling in rats given fructose (15%) in drinking water from weaning to adulthood. Wistar albino rats (4 weeks old) were randomly divided into normal control (NC), fructose control (FC), and treatment (AMT) groups and were fed for a period of 8 weeks the following diets: chow + water, chow + fructose (15%), and chow + fructose (15%) + AM (500 mg/kg per day, p.o.), respectively. Compared with the NC group, the FC group was found to have significantly (p < 0.05) raised levels of fasting blood glucose, lipid, visceral mass, plasma insulin and leptin, glycogen, and gluconeogenesis enzyme but decreased glycolytic enzyme activity. Raised levels of glucose transporter 2 protein but decreased activity of phosphatidylinositol-3-kinase (PI3K/Akt) and Janus kinase - signal transducer and activator of transcription-3 (JAK-STAT3) in hepatic tissue indicate a state of insulin and leptin resistance in the FC group. A significant (p < 0.05) lowering of physical and glycemic parameters, strengthening of the hepatic glycolytic pathway over the gluconeogenic pathway, and upregulation of the PI3K/Akt and JAK-STAT3 pathways was observed in the AMT group, as compared with the FC group. For the first time, the mechanism underlying the development of insulin resistance syndrome is delineated here, along with the potential of A. marmelos to impede it.