In vitro analysis of synergistic effect of honey against chemically induced hepatic insult in Balb/c mice.

Human being knows honey since thousands of years for its nutritional and medicinal values. Traditional medicine systems like Ayurveda have elaborated honey as boon for health and patient care. Synergistic effect of honey was research but to a limited extend. Honey is advised with Picrorhiza kurroa for hepatic disorders or hepato-protection in Ayurveda. Here we have examined in vitro synergistic effect of honey when given with Picrorhiza kurroa in acetaminophen-induced hepatotoxicity in Balb/c mice model. We obtained Serum and liver lysate to check levels of hepatic markers, Alkaline phosphatase and Glutathione. The level of alkaline phosphatase raised while glutathione level is reduced during hepatic offence. We observed a compensation of above markers when P. Kurrua, honey and its combination were used in acetaminophen induced hepatic toxicity. We observed a better mice weight gain in combination group (Group IV) compared to control group. This study can pave a way for future research on honey as a better adjuvant for hepatotoxic drugs and other herb-drug interactions researches.

In-depth assembly of organ and development dissected Picrorhiza kurroa proteome map using mass spectrometry

Background Picrorhiza kurroa Royle ex Benth. being a rich source of phytochemicals, is a promising high altitude medicinal herb of Himalaya. The medicinal potential is attributed to picrosides i.e. iridoid glycosides, which synthesized in organ-specific manner through highly complex pathways. Here, we present a large-scale proteome reference map of P. kurroa, consisting of four morphologically differentiated organs and two developmental stages. Results We were able to identify 5186 protein accessions (FDR < 1%) providing a deep coverage of protein abundance array, spanning around six orders of magnitude. Most of the identified proteins are associated with metabolic processes, response to abiotic stimuli and cellular processes. Organ specific sub-proteomes highlights organ specialized functions that would offer insights to explore tissue profile for specific protein classes. With reference to P. kurroa development, vegetative phase is enriched with growth related processes, however generative phase harvests more energy in secondary metabolic pathways. Furthermore, stress-responsive proteins, RNA binding proteins (RBPs) and post-translational modifications (PTMs), particularly phosphorylation and ADP-ribosylation play an important role in P. kurroa adaptation to alpine environment. The proteins involved in the synthesis of secondary metabolites are well represented in P. kurroa proteome. The phytochemical analysis revealed that marker compounds were highly accumulated in rhizome and overall, during the late stage of development. Conclusions This report represents first extensive proteomic description of organ and developmental dissected P. kurroa, providing a platform for future studies related to stress tolerance and medical applications.

Capturing Acyltransferase(s) Transforming Final Step in the Biosynthesis of a Major Iridoid Glycoside, (Picroside-II) in a Himalayan Medicinal Herb, Picrorhiza kurroa

BackgroundPicrorhiza kurroa has been reported as an age-old ayurvedic hepatoprotection to treat hepatic disorders due to the presence of iridoids such as picroside-II (P-II), picroside-I, and kutkoside. The acylation of catalpol and vanilloyl coenzyme A by acyltransferases (ATs) is critical step in P-II biosynthesis. Since accumulation of P-II occurs only in roots, rhizomes and stolons, uprooting of this critically endangered herb has been the only source of this compound. Recently, we reported that P-II acylation likely happen in roots, while stolons serve as the vital P-II storage compartment. Therefore, developing an alternate engineered platform for P-II biosynthesis require identification of P-II specific AT/s.Methods and results In that direction, egg-NOG function annotated 815 ATs from de novo RNA sequencing of tissue culture based ‘shoots-only’ system and nursery grown shoots, roots, and stolons varying in P-II content, were cross-compared in silico to arrive at ATs sequences unique and/or common to stolons and roots. Verification for organ and accession-wise upregulation in gene expression of these ATs by qPCR has shortlisted six putative ‘P-II-forming’ ATs. Further, six-frame translation, ab initio protein structure modelling and protein-ligand molecular docking of these ATs signified one MBOAT domain containing AT with preferential binding to the vanillic acid CoA thiol ester as well as with P-II., implying that this could be potential AT decorating final structure of P-II. ConclusionOrgan-wise comparative transcriptome mining coupled with reverse transcription real time qPCR and protein-ligand docking led to the identification of an acyltransferases, contributing to the final structure of P-II.

Ayurvedic Medicinal Plants Against COVID-19: An In Silico Analysis

Even after one and a half years since the outbreak of COVID-19, its complete and effective control is still far from being achieved despite vaccination drives, symptomatic management with available drugs, and wider lockdowns. This has inspired researchers to screen potential phytochemicals from medicinal plants against SARS-CoV-2, adopting a bio-informatics approach. The current study aimed to assess anti-viral activity of the phytochemicals derived from Ayurvedic medicinal plants against SARS-CoV-2 drug targets [3-chymotrypsin-like protease (3CLpro) and RNA dependent RNA polymerase (RdRp)] using validated in silico methods.3D Structures of 196 phytochemicals from three Ayurvedic plants were retrieved from PubChem and KNApSAcK databases and screened for Absorption Distribution Metabolism Excretion and Toxicity(ADMET) to predict drug-likeness. The phytochemicals were subjected to molecular docking and only three showed promise: Acetovanillonewith a binding affinity of −4.7Kcal/mol with RdRp and −4.1 Kcal/mol with 3CL pro; myrtenol with equivalent values of −4.3 Kcal/mol with RdRP and −3.2 Kcal/mol with 3CLpro; and nimbochalcin with equivalent values of −5.0Kcal/mol with RdRp and −4.9 Kcal/mol with 3CLpro. Molecular dynamics simulation (50ns) analysis was made of 3CLpro and RdRp using Autodock Vina 1.1.2 software and VMD software. After ADMET analysis, 78 phytochemicals were found suitable for molecular docking. Three, namely acetovanillone, myrtenol and nimbochalcin from Picrorhiza kurroa, Azadirachta indica and Cyperus rotundus,respectively,exhibited good binding affinity with 3CLproand RdRp of SARS-CoV-2. Interaction analysis, molecular dynamics simulations and MM-PBSA calculations were executed for two complexes, acetovanillone_RdRp and myrtenol_3CL pro.Acetovanillone_RdRpcomplex did not display any structural change after MD simulation as compared to myrtenol_3CL pro. The overall stability of acetovanillone_6NUR was 154.7 kJ/mol, and for myrtenol_1UJ1 90.5 kJ/mol. In silico analysis revealed that acetovanillone ( Picrorhiza kurroa) and myrtenol ( Cyperus rotundus) possess anti SARS-CoV-2 activity. Further studies are needed to validate their efficacy in biological models.

Green synthesis, characterization and antimicrobial activities of Copper nanoparticles from the rhizomes extract of Picrorhiza kurroa

Background: Green synthesized nanoparticles from the solvent extract of various plant parts show better biological activities as compared to parent solvent plant extract. Traditionally rhizomes of Picrorhiza kurroa are used to cure various diseases like diarrhea, fever, jaundice, eye infection, skin problems, asthma, arthritis, cancer, diabetes, gastrointestinal problems. Objectives: Present study describes the synthesis of copper nanoparticles from a hydroethanolic extract of P. kurroa rhizomes (CuNPs-Pk) and their evaluation for antimicrobial activities against gram-negative, gram-positive bacterial, and fungal strains. Methods: The solution of copper sulfate and hydroethanolic extract of rhizomes of P. kurroa was mixed with help of a magnetic stirrer at 60°C temperature for 1 h. The blue color of CuSO4.5H2O changed to brownish-black colored copper nanoparticles within 10 minutes. These nanoparticles were centrifuged at 4000 rpm for 20 min, washed with ethanol, followed by deionized water, dried, and were characterized by Ultra violet–visible (UV-Vis) absorption spectra, Fourier transform infrared spectroscopy (FTIR), Field emission scanning electron microscopy (SEM) and high-resolution transmission electron microscopy (TEM). Different concentrations of hydroethanolic extract of Picrorhiza kurroa rhizomes (HEEPk), CuNPs-Pk and copper oxide nanoparticles (bare CuO) ranging from 100-400 ppm had been studied against selected bacterial and fungal strains by using the well plate diffusion method. Ciprofloxin and fluconazole were used as standard and Dimethyl sulfoxide (DMSO) as a control for selected strains. Results: The UV–Vis spectral studies confirmed the surface plasmon resonance of green-synthesized CuNPs-Pk. The particle size was found to be 275-285 nm. FTIR analysis of biosynthesis nanoparticles have been confirm the presence of various functional groups (flavonoids, glycosides, tannins, phenols). SEM and TEM of biosynthesized nanoparticles have predicted their spheric al shape and their size (20-40 nm) and These particles have shown effective antimicrobial activities against selected pathogenic organisms viz. Escherichia coli, Staphylococcus aureus, and Aspergillus niger than that of HEEPk and bare CuO. Conclusions: The CuNPs-Pk show effective antimicrobial activities against bacterial and fungal pathogens as compared to HEEPk and bare CuO.

Antimicrobial Property of Picrorhiza kurroa Against Multi-Drug Resistant Uropathogenic Escherichia coli

As per the current scenario of weakened immune systems in humans and inadequate usage of antibiotics, the infections came out as serious medical conditions and pathogens, especially bacteria causing urinary tract infections to have gained resistance. To combat this, the bioactive and antimicrobial properties of healing herbs have gained the limelight. Likewise, Picrorhiza kurroa, a famous traditional and local drug in the higher Himalayan region, is used for antimicrobial treatments and partial sanitation of air by the tribes, including Ayurveda scientists. This study was done on the urine sample of a 78-year-old UTI patient, the successive pathological tests revealed the acute infected condition, and further microbiological analysis of the urine sample diagnosed the presence of pathogenic E. coli in large numbers, which was further detected to be multiple drug-resistant (MDR) by antibiotic sensitivity tests. The plant extract through the methanol extraction method of Picrorhiza kurroa roots was subjected to antimicrobial assay against the isolated pathogen E. coli by well diffusion method. The MDR E. coli was found to be highly sensitive against the plant extract indicating 23 mm microbial inhibition zone, which proves the description of Kedar Jadi as an antimicrobial drug mentioned in Charak Samhita; furthermore, this can lead to the preparation of drugs and addition in combating against acute MDR bacterial infections.