Prerna Raina

Evaluating the anti-inflammatory potential of Tectaria cicutaria L. rhizome extract in vitro as well as in vivo

ETHNOPHARMACOLOGICAL RELEVANCE The rhizome of Tectaria cicutaria has been used in the folklore system of Indian traditional medicine (Ayurveda) for the treatment of various disorders such as rheumatic pain, chest complaints, burns, sprain, poisonous bites, tonsilitis, toothache, gum complaints, cuts and wounds. The present work has for the first time tried to elucidate the anti-inflammatory potential of aqueous extract of Tectaria cicutaria rhizome (TCRaq) in vitro as well as in vivo. MATERIALS AND METHODS Anti-inflammatory potential of TCRaq was analyzed in vivo in carrageenan induced rat paw edema model. Serum antioxidant status in TCRaq-treated as well as untreated control rodents was measured by oxygen radical absorbance capacity (ORAC) assay. In vitro experiments for analyzing the anti-inflammatory potential of TCRaq were performed on murine macrophage cell line, RAW 264.7. Analysis of nitric oxide release in RAW 264.7 cells was done by Griess reaction. RT-PCR and western blotting experiment was performed to analyze the expression of iNOS. Expression of COX-2 and NFκB proteins was evaluated by western blotting. RESULTS TCRaq significantly reduced the paw volume in Sprague-Dawley rats at a dose of 200mg/kg body weight, which was comparable with the standard diclofenac treatment. The rats treated with TCRaq showed a significant increase in the serum antioxidant levels compared to the untreated control animals. TCRaq was able to reduce the nitric oxide (NO) levels in RAW 264.7 cells that had been stimulated with lipopolysaccharide (LPS). This was accompanied by a corresponding decrease in iNOS expression at mRNA and protein level. Interestingly, TCRaq was found to decrease the expression of COX-2 as well as the nuclear translocation of NFκB in RAW 264.7 cells. CONCLUSION Our study signifies the anti-inflammatory potential of Tectaria cicutaria and scientifically validates its traditional use in inflammatory conditions.

Evaluation of subacute toxicity of methanolic/aqueous preparation of aerial parts of O. sanctum in Wistar rats: Clinical, haematological, biochemical and histopathological studies.

ETHNOPHARMACOLOGICAL RELEVANCE Ocimum sanctum, commonly known as Holy Basil or Tulsi has been used in Ayurveda as a demulcent, stimulant, expectorant; in the treatment of bronchitis, skin infections, malaria, diarrhoea, dysentery, arthritis, gastric and inflammatory disorders. We have previously shown that methanolic/aqueous extract of O. sanctum did not induce genotoxicity and other toxic effects in acute oral toxicity study. In the present report, we have performed sub-acute toxicity of methanolic/aqueous preparation of O. sanctum in Wistar rats to evaluate whether it induced any chronic toxic effects. MATERIALS AND METHODS In subacute toxicity study, animals received O. sanctum extract (OSE) by oral gavage at the doses of 250, 500 and 1000 mg/kg/day (n=5/group/sex) for 28 days. At the end of the study, the animals were sacrificed and evaluated for the effect of OSE on clinical, haematological, biochemical and histopathological parameters. RESULTS The rats treated with OSE did not show any change in body weight, food and water consumption, motor activity, sensory reactivity and foot splay measurements. There were no significant changes in haematological, pathological and biochemical parameters; and histopathology of tissues (liver, kidney, spleen, heart, and testis/ovary) among rats of either sex. OSE at a dose of 1000 mg/kg showed significant increase of Mean corpuscular hemoglobin (MCH) (19.8 ± 0.8; 18.7 ± 0.5) and mean corpuscular hemoglobin concentration (MCHC) (41.8 ± 1.1; 39.3 ± 0.7) in male and female rats in comparison to their respective controls (MCH: 17.7 ± 0.3; 17.4 ± 0.3; MCHC: 37.8 ± 0.5; 36.1 ± 0.2). Urine parameters (appearance, blood, nitrate, leucocyte, glucose, ketone, pH, protein and specific gravity) in both the male and female rats were comparable to their respective controls. In addition, no changes were observed in the vital organs of rats at macroscopic and microscopic levels. CONCLUSIONS Our results showed that oral administration of OSE was not toxic to male and female Wistar rats upto the highest dose tested, thereby suggesting its clinical usefulness.

Triphala, regulates adipogenesis through modulation of expression of adipogenic genes in 3T3-L1 Cell line

Background: Triphala, an Ayurvedic polyherbal formulation, is used for the treatment of various diseases including obesity. Objective: The present study was planned to evaluate the anti-adipogenic potential of aqueous extract of Triphala (TPaq) using 3T3-L1 adipocyte cell line model. Methods: The effect of aqueous extract of Triphala (TPaq) was tested on the viability of 3T3- L1 cells by MTT assay. The cells were treated with a cocktail of dexamethasone (DEX), isobutylmethylxanthine (IBMX) and insulin to induce adipogenesis. The cells were treated either with the induction cocktail or with the cocktail containing different concentrations (1, 10 and100 μg/ml) of TPaq. Intracellular lipid content was analyzed using Oil O Red stain and was quantified after extracting with isopropanol at 500 nm wavelength. The expression of early (PPAR-γ and C/EBP-α) and late (GLUT4 and FAS) phase adipogenic genes was studied by real time PCR. Results: TPaqdid not affect the viability of 3T3-L1 cell line. Interestingly, TPaqinduced a concentration dependant decrease in the intracellular lipid content and expression of both early and late phase adipogenic genes. This decrease was statistically significant compared to cells treated with only induction cocktail. Conclusion: These results suggested that Triphala regulated lipid accumulation by down regulating expression of adipogenic genes, resulting into prevention of adipogenesis. Abbreviations used: TPaq: Aqueous extract of Triphala; DMEM: Dulbeccos Modified Eagles medium; FBS: Fetal Bovine Serum; IBMX: Isobutyl methylxanthine; DMX: Dexamethasone; MTT: [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide] assay; PPARγ: Peroxisome proliferator-activated receptor; C/EBP:Enhancer binding protein α, FAS:Fatty acid synthase; Glut-4: Glucose phosphate transporter 4.

Cinnamaldehyde, cinnamic acid, and cinnamyl alcohol, the bioactives of cinnamomum cassia exhibit HDAC8 inhibitory activity : an in vitro and in silico study

Background: The altered expression of histone deacetylase family member 8 (HDAC8) has been found to be linked with various cancers, thereby making its selective inhibition a potential strategy in cancer therapy. Recently, plant secondary metabolites, particularly phenolic compounds, have been shown to possess HDAC inhibitory activity. Objective: In the present work, we have evaluated the potential of cinnamaldehyde (CAL), cinnamic acid (CA), and cinnamyl alcohol (CALC) (bioactives of Cinnamomum) as well as aqueous cinnamon extract (ACE), to inhibit HDAC8 activity in vitro and in silico. Materials and Methods: HDAC8 inhibitory activity of ACE and cinnamon bioactives was determined in vitro using HDAC8 inhibitor screening kit. Trichostatin A (TSA), a well-known anti-cancer agent and HDAC inhibitor, was used as a positive control. In silico studies included molecular descriptor Analysis molecular docking absorption, distribution, metabolism, excretion, and toxicity prediction, density function theory calculation and synthetic accessibility program. Results: Pharmacoinformatics studies implicated that ACE and its Bioactives (CAL, CA, and CALC) exhibited comparable activity with that of TSA. The highest occupied molecular orbitals and lowest unoccupied molecular orbitals along with binding energy of cinnamon bioactives were comparable with that of TSA. Molecular docking results suggested that all the ligands maintained two hydrogen bond interactions within the active site of HDAC8. Finally, the synthetic accessibility values showed that cinnamon bioactives were easy to synthesize compared to TSA. Conclusion: It was evident from both the experimental and computational data that cinnamon bioactives exhibited significant HDAC8 inhibitory activity, thereby suggesting their potential therapeutic implications against cancer. Abbreviations used: ACE: Aqueous Cinnamon Extract; DFT: Density Function Theory; CAL: Cinnamaldehyde; CA: Cinnamic Acid; CALC: Cinnamyl Alcohol; MW: Molecular Weight; ROTBs: Rotatable Bonds; ROF: Lipinskis Rule of Five; TSA: Trichostatin A; PDB: Protein Data Bank; RMSD: Root Mean Square Deviation; HBA: Hydrogen Bond Acceptor; HBD: Hydrogen Bond Donor; ADMET: Absorption, Distribution, Metabolism, Excretion and Toxicity; FO: Frontier Orbital; HOMOs: Highest Occupied Molecular Orbitals; LUMOs: Lowest Unoccupied Molecular Orbitals; BE: Binding Energy.

Nardostachys jatamansi root extract modulates the growth of IMR-32 and SK-N-MC neuroblastoma cell lines through MYCN mediated regulation of MDM2 and p53

Aim: The present study evaluated the effect of ethanolic extract of Nardostachys jatamansi roots (NJet) on MYCN mediated regulation of expression of MDM2 and p53 proteins in neuroblastoma cell lines, IMR-32 and SK-N-MC. Materials and Methods: The effect of NJet on cell viability was determined by MTT; and on growth kinetics was evaluated by trypan blue dye exclusion method and soft agar assay. The expression of p53, MDM2 and MYCN proteins in response to NJet treatment was evaluated by immunoblotting. Results: NJet decreased the viability of neuroblastoma cells without affecting the viability of non-cancerous, HEK-293 cells. It altered the growth kinetics of the cancer cells in a dose-dependent manner. NJet down regulated the expression of MYCN and MDM2 proteins with a simultaneous increase in the expression of tumor suppressor protein p53. Conclusions: The present data demonstrated that NJet regulated the growth of IMR-32 and SK-N-MC through reduction in MYCN expression that lead to down regulation of MDM2 protein and increase in p53 expression. These preliminary results warrant further in depth studies to explore the therapeutic potential of Nardostachys jatamansi in the management of neuroblastoma. Abbreviations used: NJet: N. jatamansie thanolic extract; MTT: 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenylthiazolium bromide; FBS: Fetal bovine serum; FITC: Fluorescein isothiocyanate