Indigenous wisdom of a Kwatha to treat NASH: An insight into the mechanism.

ETHNOPHARMACOLOGICAL RELEVANCE: Nonalcoholic fatty liver disease (NAFLD) is the most common severe liver disease globally, progressing further into nonalcoholic steatohepatitis (NASH) and hepatocellular carcinoma (HCC). Vasaguduchyadi Kwatha (VK) is an Ayurvedic formulation traditionally used to treat liver diseases and other metabolic complications. This study is an ethnopharmacological approach to unravel this indigenous remedy. AIM OF THE STUDY: We aimed to discover the probable mechanism of action of VK against NASH in this study, using network pharmacology, molecular docking, in vitro study, and preclinical investigation. METHODS AND RESULTS: Among the 55 components identified, 10 were confirmed based on mass, elution charecteristics, MS/MS analysis data, and fragmentation rules. Computational study indicated 92 targets involved in the central pathways of NASH, out of which only 15 targets and 9 VK constituents have significant docking scores. In vitro and in vivo analysis results showed that VK significantly reduces weight gain and improves insulin sensitivity, dyslipidemia, steatohepatitis and overall histological features of NASH compared to saroglitazar (SGZR). CONCLUSION: Our detailed study yielded three signalling pathways related to NASH on which VK has maximum effect, bringing up a probable alternative treatment for NASH.

Antiviral Activity of Cinchona officinalis, a Homeopathic Medicine, against COVID-19.

BACKGROUND: Coronavirus disease 2019 (COVID-19) is a potentially fatal disease caused by the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Several studies have shown that hydroxychloroquine (HCQ) significantly inhibits SARS-CoV-2 infections in vitro. OBJECTIVE: Since the phytoconstituents of Cinchona officinalis (CO) are similar to those of HCQ, the objective of this study was to test the antiviral potential of different homeopathic formulations of CO. METHODS: An analysis of the molecular composition of CO was carried out using ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry, followed by a detailed docking study. The constituents of CO were docked against various targets of SARS-CoV-2, and the binding potential of the phytoconstituents was compared and quantified. The ligand with the lowest Glide docking score is considered to have the best binding affinity. The cytotoxicity of several homeopathic formulations, including CO mother tincture (CO-MT), was also checked on VeroE6 cells. A known antiviral, remdesivir, was used as a positive control for the in vitro assays to evaluate the effects of CO-MT against SARS-CoV-2-infected VeroE6 cells. RESULTS: Molecular docking studies showed that constituents of CO exhibited binding potential to various targets of SARS-CoV-2, including Mpro, PLpro, RdRp, nucleocapsid protein, ACE2 (in host) and spike protein. Quinoline, one of the constituents of CO, can potentially bind the spike protein of SARS-CoV-2. Quinic acid showed better binding capabilities with Mpro, PLpro RdRp, nucleocapsid protein and ACE2 (allosteric site) than other constituents. Quinidine exhibited better binding to ACE2. Compared to HCQ, other phytoconstituents of CO had the equivalent potential to bind the RNA-dependent RNA polymerase, nucleocapsid protein, Mpro, PLpro and spike protein of SARS-CoV-2. In vitro assays showed that homeopathic CO-MT was not cytotoxic and that CO-MT and remdesivir respectively caused 89% and 99% inhibition of SARS-CoV-2 infection in VeroE6 cells. CONCLUSION: Based on this in silico and in vitro evidence, we propose CO-MT as a promising antiviral medicine candidate for treating COVID-19. In vivo investigation is required to clarify the therapeutic potential of CO-MT in COVID-19.

Meal Ingestion of Ceraceomyces tessulatus Strain BDM-X (Agaricomycetes) Protects against Nonalcoholic Steatohepatitis in Mice.

Nonalcoholic steatohepatitis (NASH) is becoming the most common cause of hepatocellular carcinoma (HCC) in developed countries. Oxidative stress plays a major role in the pathogenesis of NASH due to steatosis; hence, novel therapeutic approaches might include natural antioxidants. Ceraceomyces tessulatus strain Basidiomycetes-X (BDM-X), a novel edible mushroom, possesses potent antioxidant activity. This study aimed to investigate the hepato-protective effect of C. tessulatus BDM-X in a novel NASH-HCC mouse model. To prepare this animal model, 2-day-old C57BL/6J male pups were exposed to low-dose streptozotocin (STZ); at 4 weeks of age, they were randomly divided into two groups. The NASH group (NASH) received a high-fat diet (HFD32) up to 14 weeks of age; the C. tessulatus BDM-X group (BDM-X) received HFD32 up to age 10 weeks, followed by HFD32 + 20% BDM-X (percent weight per weight in the diet) up to age 14 weeks. Mice not treated with STZ and fed a normal diet served as a control group. We found that C. tessulatus BDM-X improved serum aminotransferase levels as well as histopathological features such as steatosis, inflammatory foci, and pericellular fibrosis in NASH mice. Hepatic protein expression of sterol regulatory element binding protein isoform SREBP-1 and peroxisome proliferator-activated receptor PPARα was significantly increased in NASH mice. C. tessulatus BDM-X treatment normalized the expression of both proteins. Our data suggest that C. tessulatus BDM-X may protect the liver against lipogenesis in NASH-HCC mice.

Evaluation of a natural compound extracted from Dolichandrone atrovirens as a novel antioxidant agent using Caenorhabditis elegans.

The compound methyl cinnamoyl catalpol (DAM-1) was isolated from the methanol extract of Dolichandrone atrovirens. Studies have already reported the antioxidant activity of Dolichandrone atrovirens bark extract, but till date the antioxidant activity of the isolated compound DAM-1, remains unexplored. The endogenous process of reactive oxygen species generation which leads to various degenerative diseases, can be broken down using these exogenous moieties from plant origin, herein this study we sought to evaluate the antioxidant potential of the DAM-1 compound using Caenorhabditis elegans (C. elegans), which is the primary model to study the antioxidant activity of compounds. Cytotoxicity assay results showed that DAM-1 treatment in the concentration of 10, 25 and 50 µg/ml has shown 100%, 91%, and 50% survival respectively with overall p<0.0001 (treatment v/s control group). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide-Formazan (MTT) assay results showed that treatment had better survival rates than the control group at different time intervals i.e. 48 h, and 72 h with p<0.01. Mechanosensation (behavioral study) as well as in vivo study results showed that at 0 h, 10 µg/ml of DAM-1 treatment showed a better anti-oxidative activity than the control group, 25 and 50 µg/ml of DAM-1 treated groups with p<0.001 but at 2.5 h incubation with 10, 25, 50 µg/ml of DAM-1 showed an increased anti-oxidative activity than the control group with p<0.001. Thermoresistance assay confirmed that the treatment group had more survival than control group with p<0.001. Absorption study of DAM-1 in C. elegans has shown that the absorption of the drug increases up to 180 mins with a slight decrease after 360 mins and then constant absorption up to 1440 mins. This study paves the way towards the initiative to explore the pharmacological role of DAM-1 in various oxidative stress mediated diseases at molecular levels and the absorption study points out its potential role which could be utilized in the metabolomics and proteomics analysis of this compound in other studies.