Comparative spasmolytic effect between Cinnamomum tamala and Cinnamomum verum leaf essential oils and eugenol through in vitro and in silico approaches.

Cinnamomum tamala and Cinnamomum verum are known for their folk medicinal usage in treating gastrointestinal ailments. The spasmolytic activity of essential oils was studied using isolated rat ileum. The results indicate that C. tamala, despite having a lower content of eugenol (60%), shows a spasmolytic potential of 68.01 ± 2.63% (EC50 = 110.12 ± 13.58 µg/mL) while C. verum with rich eugenol (80%) shows lesser spasmolytic potential (38.96 ± 0.63%) and fails to attain an EC50 value. Upon comparison with standard eugenol's percentage of spasmolytic (35.68 ± 2.57%), it is evident that the action of these essential oils does not solely rely on the major component but the synergistic role in association with other components of the mixture influences the pharmacological action of the essential oils. In silico docking of phytochemicals of leaf essential oils with M2 (M2AChR) and M3 muscarinic (M3AChR) and nicotinic acetylcholine receptor (nAChR) was carried out to determine the type of receptors through which the essential oils had spasmolytic potential. The binding affinity for eugenol with nAChR shows a better docking score than M2AChR and M3AChR.

Vasorelaxant property of Plectranthus vettiveroides root essential oil and its possible mechanism.

ETHNOPHARMACOLOGICAL RELEVANCE: Plectranthus vettiveroides (Jacob) N.P. Singh & B.D. Sharma is a traditional medicinal plant used in Siddha System of Medicine and its aromatic root is used to reduce the elevated blood pressure. AIM: The aim of the present study was to study vasorelaxant property of the root essential oil nanoemulsion (EON) of P. vettiveroides. METHODS: The EON was formulated to enhance the solubility and bioavailability and characterized. The preliminary screening was performed by treating the EON with aortic rings pre-contracted with phenylephrine (1 µM) and potassium chloride (80 mM). The role of K⁺ channels in EON induced vasorelaxation was investigated by pre-incubating the aortic rings with different K⁺ channel inhibitors namely, glibenclamide (a non-specific ATP sensitive K⁺ channel blocker, 10 µM), TEA (a Ca2⁺ activated non-selective K⁺ channel blocker, 10-2 M), 4-AP (a voltage-activated K⁺ channel blocker, 10-3 M) and barium chloride (inward rectifier K⁺ channel blocker, 1 mM). The involvement of extracellular Ca2+ was performed by adding cumulative dose of extracellular calcium in the presence and absence of EON and the concentration-response curve (CRC) obtained is compared. Similarly, the role of nitric oxide synthase, muscarinic and prostacyclin receptors on EON induced vasorelaxation were evaluated by pre-incubating the aortic rings with their inhibitors and the CRC obtained in the presence and absence of inhibitor were compared. RESULTS: The GC-MS and GC-FID analyses of the root essential oil revealed the presence of 62 volatile compounds. The EON exhibited significant vasorelaxant effect through nitric oxide-mediated pathway, G-protein coupled muscarinic (M3) receptor pathway, involvement of K+ channels (KATP, KIR, KCa), and blocking of the calcium influx by receptor-operated calcium channel. CONCLUSION: It is concluded that the root essential oil of P. vettiveroides is possessing marked vasorelaxant property. The multiple mechanisms of action of the essential oil of P. vettiveroides make it a potential source of antihypertensive drug.

Investigations on a polyherbal formulation for treatment of cognitive impairment in a cholinergic dysfunctional rodent model.

Alzheimer's disease is a multifactorial neurodegenerative condition manifested through acute cognitive decline, amyloid plaque deposits and neurofibrillary tangles. Complete cure for this disease remains elusive as the conventional drugs address only a single molecular target while Alzheimer's disease involves a complex interplay of different sets of molecular targets and signaling networks. In this context, the possibility of employing multi-drug combinations to rescue neurons from the dysregulated metabolic changes is being actively investigated. The present work investigates a poly-herbal formulation, Brahmi Nei that has been traditionally used for anxiolytic disorders and immunomodulatory effects, for its efficiency in ameliorating cognitive decline through a combination of behavioral, biochemical, histopathological, gene and protein expression analyses. Our results reveal that the formulation shows excellent neuroregenerative properties, rescues neurons from inflammatory damage, reduces neuritic plaque deposits and improves working memory in rodent models with scopolamine-induced dementia. The microarray analysis shows that the formulation induces the expression of pro-survival pathways and positively modulates genes involved in memory consolidation, axonal growth and proliferation in a concentration-dependent manner with therapeutic concentrations restoring the normal conditions in the brain of the diseased animals. The neuritic spine morphology confirms the long-term memory potentiation through improved mushroom spine density, increased dendritic length and connectivity. Taken together, our study provides mechanistic evidence to prove that the traditional formulation can be a superior therapeutic strategy to treat cognitive decline when compared to the conventional mono-drug treatment.

Microarray analysis of genes from animals treated with a traditional formulation ChandraprabhaVati reveals its therapeutic targets.

BACKGROUND: Traditional medicinal preparations have not received global acceptance, and their therapeutic benefits remain disputed due to lack of scientific evidence on their mechanism of action. Microarray analysis has emerged as a powerful technique that can aid in understanding the complex signaling networks activated by these formulations and thereby assess their beneficial as well as adverse effects. AIM: The present work aims to investigate the differential influence of ChandraprabhaVati, Ayurvedic formulation used in the treatment of diabetes, anemia, urinary, respiratory, skin and liver disorders. MATERIALS AND METHODS: The RNA from the liver of rats treated with different doses of ChandraprabhaVati for 28 days was isolated and studied for the genome-wide changes in the expression. RESULTS: The results revealed several molecular targets that could contribute to the therapeutic effects of ChandraprabhaVati. Several genes have been differentially expressed, among those miRNAs miR-434, miR877, and miRlet7e contribute to the anti-diabetic, anti-fibrotic and anti-inflammatory of CPV. The rejuvenative activity of CPV may be due to the MeOX1 and Upf3b genes. Up-regulation of Hbaa2 gene facilitates the anti-anemic effect. Interestingly gender-specific differential expressions of genes were also observed. Rab3d were found to be altered in female when compared to male animals. CONCLUSION: Thus the microarray data for the CPV treated animals has revealed molecular targets that may be responsible for the various known therapeutic effects and also identified new beneficial effects of CPV.

Vasorelaxant and cardiovascular properties of the essential oil of Pogostemon elsholtzioides.

ETHNOBOTANICAL RELEVANCE: Pogostemon elsholtzioides Benth. (Lamiaceae) is an aromatic shrub, endemic to eastern Himalaya region. The leaves are used for treating goiter and high blood pressure (BP) by indigenous people in Arunachal Pradesh, India. Young leaves are used as vegetable and leaf decoction is also used for cough, cold and headache by some indigenous communities in Northeast India. AIM OF THE STUDY: This species is used for treating hypertension and the genus Pogostemon is rich in essential oil. Therefore, the present study was aimed at investigation of the chemical constituents, vasorelaxant and cardiovascular effects of the essential oil of P. elsholtzioides. MATERIALS AND METHODS: P. elsholtzioides was collected from Pasighat, Arunachal Pradesh, India and essential oil was extracted from shade dried leaves. Essential oil was analyzed by GC-FID and GC-MS and the volatile constituents were identified. Vasorelaxant and cardiovascular properties of the essential oil were studied against phenylephrine induced contraction in isolated endothelium intact aortic preparations and by measuring systolic and diastolic BP, mean arterial pressure (MAP) and heart rate (HR) after carotid artery cannulation in Wistar rats. RESULTS: The essential oil was rich in sesquiterpenes and curzerene, benzophenone, α-cadinol and germacrone were major constituents. The essential oil exhibited significant vasodilation effect in phenylephrine induced contracted aortic rings. Vasorelaxant effect of the essential oil was also observed both in the presence and absence of Nitro-L-arginine methyl ester against phenylephrine-contracted aortic rings. It also induced reduction of systolic and diastolic BP, MAP and HR. CONCLUSIONS: Essential oil of P. elsholtzioides exhibited significant vasorelaxant effect against endothelium intact aortic preparation mediated through nitric oxide dependent pathway and also reduced BP. However, further study is needed to screen the role of calcium ions in both intracellular and extracellular pathway.

Multi-functional nanoparticles as theranostic agents for the treatment & imaging of pancreatic cancer.

Theranostics has received considerable attention since both therapy and imaging modalities can be integrated into a single nanocarrier. In this study, fluorescent iron oxide (FIO) nanoparticles and gemcitabine (G) encapsulated poly(lactide-co-glycolide) (PLGA) nanospheres (PGFIO) conjugated with human epidermal growth factor receptor 2, (HER-2) antibody (HER-PGFIO) were prepared and characterized. HER-PGFIO showed the magnetic moment of 10emu/g, relaxivity (r2) of 773mM-1s-1 and specific absorption rate (SAR) of 183W/g. HER-PGFIO showed a sustained release of gemcitabine for 11days in PBS (pH 7.4). In vitro cytotoxicity evaluation of HER-PGFIO in 3D MIAPaCa-2 cultures showed 50% inhibitory concentration (IC50) of 0.11mg/mL. Subcutaneous tumor xenografts of MIAPaCa-2 in SCID mice were developed and the tumor regression study at the end of 30days showed significant tumor regression (86±3%) in the HER-PGFIO with magnetic hyperthermia (MHT) treatment group compared to control group. In vivo MRI imaging showed the enhanced contrast in HER-PGFIO+MHT treated group compared to control. HER-PGFIO showed significant tumor regression and enhanced MRI in treatment groups, which could be an effective nanocarrier system for the treatment of pancreatic cancer. STATEMENT OF SIGNIFICANCE: Combination therapies are best suitable to treat pancreatic cancer. Theranostics are the next generation therapeutics with both imaging and treatment agents encapsulated in a single nanocarrier. The novelty of the present work is the development of targeted nanocarrier that provides chemotherapy, thermotherapy and MRI imaging properties. The present work is the next step in developing the nanocarriers for pancreatic cancer treatment. Different treatment modalities embedding into a single nanocarrier is the biggest challenge that was achieved without compromising the functionality of each other. The surface modification of polymeric nanocarriers for antibody binding and their multifunctional abilities will appeal to wider audience.

Gastroprotective potential of hydro-alcoholic extract of Pattanga (Caesalpinia sappan Linn.).

ETHNO-PHARMACOLOGICAL RELEVANCE: Pattanga is botanically equated as Caesalpinia sappan Linn. (Family: Caesalpiniaceae) and is used in Ayurveda system of medicine since ages. According to Ayurveda, useful part is Heartwood, which is bitter, astringent and acrid and is useful in vitiated conditions of vata and pitta, burning sensation, wounds, ulcers, leprosy, skin diseases, menorrhagia, leucorrhea, and diabetes. It is used as a major ingredient in Ayurvedic formulations and preparations like Patrangasava, Chandanadya Thalia, and Karpuradyarka. AIM OF THE STUDY: The present study is planned to evaluate the gastroprotective activity of the selected Ayurvedic drug using three different in vivo gastric ulcer models, so as to provide scientific evidence for the Ayurvedic claims. MATERIALS AND METHODS: For this study, Wistar albino rats fasted overnight were selected. The hydroalcoholic extract of Caesalpinia sappan heartwood at the dose level 250 and 500mg/kg body weight was selected and administered orally before necrotizing agents. Antioxidant and antiulcer parameters were evaluated and the stomach samples were subjected for histopathological studies. In addition, PGE2 estimation and protein expressions of COX-1, COX-2 and iNOS were analyzed by Western blot. The plant extract was subjected to LCMS/MS analysis. In addition, Cytoprotective effect in isolated gastric mucosal cells, TUNEL Assay, Acid neutralizing capacity assay, H+/K+ ATPase inhibitory assay were performed. RESULTS: The ulcer protection was found to be 92%, 86% and 64% against ethanol, NSAID and pylorus ligation induced ulcer respectively. The hydro-alcoholic extract of C. sappan heartwood exhibited cytoprotective effect with 76.82% reduction against indomethacin-induced cytotoxicity at the concentration of 25µg/ml. C. sappan showed 63.91% inhibition in H+/K+ ATPase inhibitory assay at the concentration 500µg/ml. CONCLUSIONS: Our results depict that Caesalpinia sappan heartwood possesses gastroprotective activity, possibly mediated through cytoprotection and antioxidant mechanisms. The data obtained in the present study provides scientific support for the traditional use of Caesalpinia sappan in the management of peptic ulcer.

Antidiabetic, antihyperlipidaemic, and antioxidant activity of Syzygium densiflorum fruits in streptozotocin and nicotinamide-induced diabetic rats.

Context Syzygium densiflorum Wall. ex Wight & Arn (Myrtaceae) has been traditionally used by local tribes of the Nilgiris, Tamil Nadu, India, for the treatment of diabetes, however, no definitive experimental studies are available. Objective This study investigates the antidiabetic, antihyperlipidaemic and antioxidant activities of ethanol extract of S. densiflorum (EFSD) fruits in streptozotocin (STZ) and nicotinamide (NA)-induced diabetic rats. Materials and methods Acute oral toxicity and oral glucose tolerance were assessed in normal rats. The antidiabetic, antihyperlipidaemic and antioxidant activities were investigated in STZ - NA-induced diabetic rats. Diabetic rats were orally administered with glibenclamide (10 mg/kg b.wt), EFSD (200, 400 and 800 mg/kg b.wt) for 28 d. Further, changes in the blood glucose level (BGL), biochemical parameters, antioxidants were observed and histology of pancreas was performed. Results No toxicity and lethality were observed. Results of the following parameters are represented by treated versus disease control (STZ + NA) groups. BGL (161.33 ± 22.8 versus 476.17 ± 56.58 mg/dl), glycosylated haemoglobin (5.285 ± 0.19 versus 8.05 ± 0.55%), urea (40.32 ± 1.96 versus 75.37 ± 2.91 mg/dl), uric acid (1.2 ± 0.07 versus 2.16 ± 0.05 mg/dl), total cholesterol (89.3 ± 5.14 versus 139.7 ± 5.95 mg/dl) and triglycerides (79.65 ± 2.52 versus 108.9 ± 3.61 mg/dl) were significantly decreased, whereas haemoglobin (11.75 ± 0.73 versus 7.95 ± 0.42 g/dl), high-density lipoprotein cholesterol (14.2 ± 1.11 versus 6.97 ± 0.84 mg/dl), total protein (45%) and liver glycogen (87%) were significantly increased in EFSD-treated diabetic group. Significant changes were observed in the enzymatic and non-enzymatic antioxidants in EFSD-treated groups (p < 0.001). Histopathological examination showed the regeneration of ß-cells in Islets of Langerhans. Conclusion This study confirms the antidiabetic, antihyperlipidaemic and antioxidant activities of S. densiflorum fruits.

Pharmacognostical, antioxidant and antiulcer screening of Cyclea peltata roots

Cyclea peltata Hook. f. & Thomson, belongs to the family Menispermaceae commonly known as "paatha", is a climbing shrub found throughout South and East India. In Indian traditional medicine the tuberous roots are also used to treat jaundice, stomachache, fever and asthma. This study deals with the microscopic study of leafs and roots of C. peltata, along with the physico-chemical, preliminary phytochemical analyses and antioxidant properties. Acute oral toxicity study was conducted as per OECD-423 guidelines and the extract was found to be devoid of any conspicuous acute toxicity in extract treated animals and no mortality upto 2 g/kg by oral route. Hydroalcoholic extract of C. peltata in a dose dependant manner (125 and 250 mg/kg. p.o.) showed significant gastric protection against the ethanol-induced gastric ulcer model in rats.

Antioxidant activity of Nelumbo nucifera (Gaertn) flowers in isolated perfused rat kidney / Atividade antioxidante das flores de Nelumbo nucifera (Gaertn) em rim isolado perfundido de rato

A padronização da indução de estresse oxidativo com a mstura de Fenton em rim isolado perfundido de rato e a avaliação do efeito antioxidante das flores de Nelumbo nucifera em rim isolado de rato oxidativamente estressado são realizadas no presente estudo. Seis grupos de rim isolado perfundido de rato foram utilizados para o presente estudo. O grupo I de rins recebeu veículo. O grupo II de rins foi oxidativamente estressados com a mistura de Fenton. Os grupos III e IV de animais foram tratados com duas doses graduais de extrato após a administração da mistura de Fenton. Os grupos V e VI de rins foram perfundidos com duas doses graduais de extrato sem a mistura de Fenton. A mistura de Fenton causou estresse oxidativo com diferença significativa de malondialdeído (TBARS), glutationa reduzida (GSH), glutationa peroxidase (GPx), catalase, glutamato oxaloacetato transaminase (GOT) e glutamato piruvato transaminase (GPT) (p<0,05) em ratos doentes. No tratamento dos animais com extrato, o estresse oxidativo foi diminuído com aumento nos antioxidantes e as enzimas marcadoras mantiveram o nível normal. Este efeito do extrato foi dependente da dose. Em conclusão, a mistura de Fenton desenvolveu o estresse oxidativo em rim isolado perfundido de rato e o extrato das flores de Nelumbo nucifera apresenta atividade antioxidante no rim isolado perfundido oxidativamente estressado.

Standardization of induction of oxidative stress with Fenton mixture in isolated perfused rat kidney and evaluation of antioxidant effect of Nelumbo nucifera flowers in isolated oxidatively stressed rat kidney are carried out in present study. Six groups of isolated perfused rat kidney were used for the present study. Group I kidneys were received vehicle. Group II kidneys were oxidatively stressed with Fenton mixture. Group III and IV animals were treated with two graded doses of extract after the administration of Fenton mixture. Group V and VI kidneys were perfused with two graded doses of extract without Fenton mixture. Fenton mixture was found to cause oxidative stress with significant difference of malondialdehyde (TBARS), Reduced glutathione (GSH), Glutathione peroxidase (GPx), catalase, Glutamate Oxaloacetate Transaminase (GOT) and Glutamate Pyruvate Transaminase (GPT) (p<0.05) in diseased rats. On treating animals with extract, the oxidative stress was decreased with increase in antioxidants and the marker enzymes were found to maintain the normal level. This effect of extract was found to be dose dependent. In conclusion, Fenton mixture developed the oxidative stress in isolated perfused rat kidney and Nelumbo nucifera flowers extract exhibits antioxidant activity in that oxidatively stressed isolated perfused kidney.