Glycyrrhizin inhibits swine acute diarrhea syndrome coronavirus infection via the HMGB1/TLR4/JNK signal pathway

Swine acute diarrhea syndrome coronavirus (SADS-CoV), a newly discovered enteric coronavirus, is the etiological agent that causes severe clinical diarrhea and intestinal pathological damage in piglets. In this study, Vero E6 and IPI-2I cells were pretreated with different concentrations of glycyrrhizin (GLY) for 2 hours, and then infected with different concentrations of SADSCoV, aiming to investigate the inhibitory effect of GLY on SADS-CoV. Western blot and TCID50 results revealed a significantly decreased N protein expression and viral titer, indicating that GLY can inhibit the infection of SADS-CoV. Vero E6 and IPI-2I cells were pretreated with different concentrations of GLY for 2 hours and infected with SADS-CoV. Western blot results showed that when the concentration of GLY was 0.8 mmol/L, the expression of N protein decreased significantly, indicating that GLY inhibited the invasion of the virus. At first, cells were treated with 0.4 mmol/L GLY, and cell samples were collected at 2 hours, 6 hours and 12 hours after being infected with SADS-CoV for analysis, and the expression of N protein were found to be significantly reduced at all points, indicating that GLY had a significant inhibitory effect on the replication of the virus. GLY is a competitive inhibitor of high mobility group box 1 (HMGB1), and the receptors of HMGB1 mainly include TLR4 and RAGE. Based on this fact, the mutant plasmid at the key sites of HMGB1 (C45S, C106S, C45/106S) and the siRNA of the RAGE receptor were transfected to Vero E6 cells and infected with SADS-CoV, and the cell supernatant and samples were harvested. The western blot and TCID50 results showed that the expression of N protein and the virus titer were decreased, suggesting that GLY exerts its function by affecting the binding of HMGB1/TLR4/RAGE during SADS-CoV infection. To further explore the signaling pathway through which GLY functions, Vero E6 and IPI-2I cells were inoculated with SADS-CoV, and cell samples were harvested, western blot was used to detect the changes of MAPK proteins. The results showed that the protein expression levels of p-p38, p-JNK and p-ERK were up-regulated in the early and late stages, indicating that the MAPK pathway was activated by SADS-CoV infection. Vero E6 and IPI-2I were pretreated with different concentrations of GLY and TLR4 inhibitor TAK for 2 hours and infected with SADS-CoV. Protein samples were harvested and analysed by western blot which showed a decreased p-JNK and N proteins, while other proteins showed no significant changes. These results indicated that GLY and TAK regulated the phosphorylation of JNK but did not regulate the phosphorylation of p38 and ERK. Also, Vero E6 cells were treated with HMGB1 antibody, the siRNA of HMGB1 and HMGB1 mutants plasmid, and infected with SADS-CoV. Protein samples were harvested, western blot results showed that phosphorylation of JNK decreased, indicating that HMGB1 affected JNK phosphorylation. Finally, Vero E6 and IPI-2I cells were pretreated with different concentrations of JNK inhibitor SP600125 to infect SADS-CoV, western blot, TCID50 and IFA results showed that the expression of N protein and virus titer, as well as virus replication were reduced, indicating that SP600125 inhibited virus replication. In conclusion, our results revealed that GLY can inhibit in vitro replication of SADS- CoV, mainly through the HMGB1/TLR4/JNK signaling pathway. The discovery of this pathway provides theoretical support for the research of novel anti-SADS-CoV drugs.

In-silico screening of plant-derived natural compounds for their anti-COVID-19 potential

The novel coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) began in Wuhan, China, in December 2019 and quickly spread across the worldwide. It becomes a global pandemic and risk to the healthcare system of almost every nation around the world. In this study thirty natural compounds of 19 Indian herbal plants were used to analyze their binding with eight proteins associated with COVID -19. Based on the molecular docking as well as ADMET analysis, isovitexin, glycyrrhizin, sitosterol, and piperine were identified as potential herbal medicine candidates. On comparing the binding affinity with Ivermectin, we have found that the inhibition potentials of the Trigonella foenum-graecum (fenugreek), Glycyrrhiza glabra (licorice), Tinospora cordifolia (giloy) and Piper nigrum (black pepper) are very promising with no side-effects.

Screening of active ingredients from traditional Chinese medicine against the novel Coronavirus based on molecular docking

ADMEN prediction was used to perform the first round screening from Traditional Chinese Medicine Database and Analysis Platform(TCMSP). then VASARA and molecular docking were used to screen again based on targets spike glyoprotein and angiotensin converting enzyme 2. and finally the interaction between target and drug was analyzed. 425 candidate ingredients of traditional Chinese medicine were screened from TCMSP database. when targeted by Spike glycoprotein. 12 ingredients were screened. They were contained artemisia apiacea salvia miltiorrhiza bge. scutellaria baicalensis. pinellia ternate. liquorice. radixImplettri and other traditional Chinese medicine. With ACE2 as the target. 77 components of traditional Chinese medicine were screened out. including salvia miltiorrhiza bge scutellaria baicalensis. pinellia ternatc. Liquorice. radix bupleuri. ephedra and other traditional Chinese medicine. At last. salviolone and dihydrotanshinlactone were found to be the potential inhibitor.

Corona virus & Ayurveda

COVID-19 is an infectious disease caused by a newly discovered CORONAVIRUS. It's a type of Severe Acute Respiratory Syndrome. The symptoms of SARS-nCOV-2 cause dry cough, fever, Tiredness, and difficulty of breathing (severe cases). We can cure the symptoms and defects of the whole body (vata, pitta, kapha) caused by the system and its qualitative therapists.

Exploring the mechanism of Platycodonis Radix-Licorice for prevention and treatment of COVID-19 based on network pharmacology and molecular docking

Objective: To investigate the possible mechanism of Platycodonis Radix-Licorice drug pair in the intervention of COVID-19 by using network pharmacology and molecular docking technique. Methods The database TCMSP was retrieved for the chemical constituents and targets of Platycodonis Radix-Licorice drug pair. Coronavirus disease targets were screened by the Gene Cards, OMIM,TTD, PharmGkb and DrugBank database. Cytoscape 3.7.2 software was used to construct the drug-component-target network. The PPI(protein-protein interaction) network was obtained by drug-disease intersection targets, and the core genes were found through CytoNCA plug-in. Meanwhile, GO(gene ontology) analysis and KEGG(Kyoto encyclopedia of genes and genomes) pathway analysis were performed by using Bioconductor database to predict the mechanism. AutoDock Tools 1.5.6 software was used to simulate the molecular docking of the main active ingredients with the novel coronavirus key binding site protein [SARS-CoV-2 main protease(severe acute respiratory syndrome coronavirus 2 main protease, Mpro) and ACE2(angiotensin converting enzyme 2)]. Results A total of 7 active ingredients of Platycodonis Radix,92 active ingredients of Licorice,2766 drug targets, and 674 disease targets were obtained, and 67 drug-disease common targets were excavated. The key targets involved RELA,STAT1,MAPK3,TP53,MAPK1,MAPK8,STAT3,MAPK14,IL1 B and TNF by the database STRING and CytoNCA plug-in.Go enrichment analysis showed that the main functions of Platycodonis Radix-Licorice drug pair on the intervention of COVID-19 were antioxidant reaction, cell respond to chemical stress, regulation of apoptotic signaling pathways, reaction to lipopolysaccharides and reaction to bacteria-derived molecules, etc.. KEGG pathways involved Coronavirus disease-COVID-19 pathway, IL-17 signaling pathway and so on, were mainly associated with immune response, inflammation-related pathways, inhibition of viral infection, and other inhibition of cancer. The molecular docking results showed that glepidotin A,quercetin, licochalcone a and luteolin had good binding ability with Mpro and ACE2. Conclusion Platycodonis Radix-Licorice drug pair act on SARS-CoV-2 through multiple components, multiple targets, and multiple channel combination. And the main active ingredients have a fine binding ability with Mpro and ACE2. The method can provide theoretical support for the possibility of traditional Chinese medicine(TCM) against COVID-19.

HYSSOP and POLIUM could help to prevent COVID-19 in high-risk population: the results of a parallel randomized placebo-controlled field trial

This study was conducted to evaluate the effect of HYSSOP (composed of Hyssopus officinalis L., Echium amoenum Fisch & C. A. Mey and Glycyrrhiza glabra L.) and POLIUM (contained Teucrium polium L., Cuscuta epithymum Murr and Cichorium intybus L.) combined distilled herbal medicines compared to placebo in the prevention of COVID-19. This is a double-blind parallel placebo-controlled field trial conducted on 751 asymptomatic individuals whose one of the family members recently had a positive RT-PCR test for COVID-19. They were divided into three groups including POLIUM, HYSSOP and placebo using random blocks with a 1:1:1 allocation ratio. Participants received daily 5 cc (under 12 years) or 10 cc (over 12 years) of allocated oral medications for 20 days. The primary outcome was the frequency of positive RT-PCR test among participants who became symptomatic. The mean age of participants was 36.6. Nineteen participants get infected by COVID-19 during the intervention;fifteen of them belonged to the placebo and four to the POLIUM group. Fisher's exact test indicated significant differences between HYSSOP and placebo (p<0.001) as well as POLIUM and placebo (p=0.009) groups in terms of COVID-19 confirmed by PCR tests. Cox regression model adjusted for confounders illustrated that the hazard of getting infection by COVID-19 in POLIUM and HYSSOP groups decreased by 66% (OR (95% CI): 0.34 (0.12 to 0.94);p=0.038) and 93% (OR (95% CI): 0.07 (0.01to 0.56);p=0.012) respectively, compared to placebo. Oral administration of HYSSOP and POLIUM with the other supportive health care could decrease the risk of getting COVID-19.

Long-Lasting Olfactory Dysfunction in Hospital Workers Due to COVID-19: Prevalence, Clinical Characteristics, and Most Affected Odorants

Hospital workers have increased exposure risk of healthcare-associated infections due to the frontline nature of their work. Olfactory dysfunction is highly prevalent. The objectives for this investigation are to study the prevalence of long-lasting olfactory dysfunction associated with COVID-19 infection in hospital workers during the first pandemic wave, to identify clinical characteristics and associated symptomatology, and to analyze how many patients with COVID-19 infection had developed olfactory dysfunction during infection and maintained a reduced olfactory function for approximately 10 weeks after diagnosis. Between June and July of 2020, a cross-sectional study was carried out at the Hospital Central de la Cruz Roja San José and Santa Adela in Madrid, Spain. One hundred sixty-four participants were included, of which 110 were patient-facing healthcare staff and 54 were non-patient-facing healthcare staff. Participants were split into three groups, according to COVID-19 diagnosis and presence of COVID-19 related olfactory symptomatology. Participants were asked to complete a structured online questionnaire along with Sniffin’ Stick Olfactory Test measurements. In this study, 88 participants were confirmed for COVID-19 infection, 59 of those participants also reported olfactory symptomatology. The prevalence of COVID-19 infection was 11.35%, and the prevalence for olfactory dysfunction was 67.05%. Olfactory dysfunction associated with COVID-19 infection leads to long-lasting olfactory loss. Objective assessment with Sniffin’ Stick Olfactory Test points to odor identification as the most affected process. Lemon, liquorice, solvent, and rose are the odors that are worst recognized. Mint, banana, solvent, garlic, coffee, and pineapple, although they are identified, are perceived with less intensity. The findings of this study confirmed a high prevalence of SARS-CoV-2 infection among the hospital workers.

Research Progress on the Antiviral Activity of Glycyrrhizin and its Derivatives in Liquorice.

Liquorice is a traditional medicine. Triterpenoids such as glycyrrhizin and glycyrrhetinic acid are the main active constituents of liquorice. Studies have revealed that these compounds exert inhibitory effects on several viruses, including SARS-CoV-2. The main mechanisms of action of these compounds include inhibition of virus replication, direct inactivation of viruses, inhibition of inflammation mediated by HMGB1/TLR4, inhibition of ß-chemokines, reduction in the binding of HMGB1 to DNA to weaken the activity of viruses, and inhibition of reactive oxygen species formation. We herein review the research progress on the antiviral effects of glycyrrhizin and its derivatives. In addition, we emphasise the significance of exploring unknown antiviral mechanisms, structural modifications, and drug combinations in future studies.

Can Antiviral Activity of Licorice Help Fight COVID-19 Infection?

The phytotherapeutic properties of Glycyrrhiza glabra (licorice) extract are mainly attributed to glycyrrhizin (GR) and glycyrrhetinic acid (GA). Among their possible pharmacological actions, the ability to act against viruses belonging to different families, including SARS coronavirus, is particularly important. With the COVID-19 emergency and the urgent need for compounds to counteract the pandemic, the antiviral properties of GR and GA, as pure substances or as components of licorice extract, attracted attention in the last year and supported the launch of two clinical trials. In silico docking studies reported that GR and GA may directly interact with the key players in viral internalization and replication such as angiotensin-converting enzyme 2 (ACE2), spike protein, the host transmembrane serine protease 2, and 3-chymotrypsin-like cysteine protease. In vitro data indicated that GR can interfere with virus entry by directly interacting with ACE2 and spike, with a nonspecific effect on cell and viral membranes. Additional anti-inflammatory and antioxidant effects of GR cannot be excluded. These multiple activities of GR and licorice extract are critically re-assessed in this review, and their possible role against the spread of the SARS-CoV-2 and the features of COVID-19 disease is discussed.

[Acute myocardial infarction and severe hypokalaemia due to liquorice consumption during COVID-19 confinement]. / Infarto agudo de miocardio e hipopotasemia severa por consumo de regaliz durante el confinamiento por COVID-19.

Liquorice is one of the oldest known herbs with medicinal properties and comprises up to 300 active compounds. It has been used for millennia for its digestive, anti-inflammatory and anti-infective properties. However, its possible toxic effects were described only a few years ago and there is growing interest in the side effects associated with chronic consumption. The main active component of liquorice is the prodrug glycyrrhizin and its active metabolite glycyrrhetic acid. It is a rare cause of hypokalaemia due to suppression of the renin-angiotensin-aldosterone axis, causing pseudohyperaldostenonism (PHA). We describe a rare case of secondary acute myocardial infarction in a patient with chronic consumption of liquorice.

Effects of Licorice on clinical symptoms and laboratory signs in moderately ill patients with pneumonia from COVID-19: A structured summary of a study protocol for a randomized controlled trial.

OBJECTIVES: We investigate the effects of Licorice (Glycyrrhiza glabra L.) root extract, an anti-inflammatory natural medicine, compared to the usual therapeutic regimen on clinical symptoms and laboratory signs in patients with confirmed COVID-19 that are moderately ill. TRIAL DESIGN: This is a single-center, open-label, randomized, clinical trial with parallel-group design. This study is being conducted at Shahid Mohammadi Hospital, Bandar Abbas, Iran. PARTICIPANTS: Both male and female patients with ≥18 years of age (≥ 35 kg of weight), admitted at the Shahid Mohammadi Hospital, Hormozgan University of Medical Sciences, Bandar Abbas for treatment, screened for the following criteria. INCLUSION CRITERIA: 1. Confirmed diagnosis of SARS-CoV-2 infection (via polymerase chain reaction [PCR] and/or antibody test). 2. Presenting as moderate COVID-19 pneumonia (via chest computed tomography (CT) and/or X-ray) requiring hospitalization. 3. Hospitalized ≤48 hours. 4. Signing informed consent and willingness of study participant to accept randomization to any assigned treatment arm. EXCLUSION CRITERIA: 1. Underlying diseases, including chronic heart disease, chronic hypertension, severe renal failure, severe liver failure, and thyroid disorders. 2. Severe and critical COVID-19 pneumonia. 3. Use of warfarin, selective serotonin reuptake inhibitors (SSRIs), monoamine oxidase inhibitors (MAOIs), diuretics, corticosteroids, and antiarrhythmic drugs. 4. Treatment with Investigational and antiviral therapy in a clinical study within one month before randomization. 5. History of allergy to Licorice. 6. Pregnancy and breastfeeding. INTERVENTION AND COMPARATOR: Intervention group: The standard treatment regimen for COVID-19 along with a Licorice-based herbal preparation (D-Reglis ®, Irandarouk Pharmaceutical Company, Iran) at a dose of 760 mg three times a day for a period of seven days. CONTROL GROUP: The standard treatment for COVID-19 based on the Iranian Ministry of Health and Medical Education's protocol for a period of seven days. MAIN OUTCOMES: The recovery rate of clinical symptoms, including fever, dry cough, and tiredness, as well as paraclinical features, including thrombocytopenia, lymphocytopenia, and C-reactive protein, are evaluated as primary outcomes within seven days of randomization. Time to improvement of clinical and paraclinical features and length of stay in a hospital, along with the incidence of adverse reactions are also evaluated as the secondary outcomes within seven days of randomization. RANDOMIZATION: An electronic table of random numbers will be used to allocate the included participants into either control or intervention groups (in a 1:1 ratio) using the simple randomization method. BLINDING (MASKING): This is an open-label trial without blinding and placebo control. NUMBERS TO BE RANDOMIZED (SAMPLE SIZE): A total of 60 participants randomizes (30 patients allocated to the intervention group and 30 patients allocated to the control group). TRIAL STATUS: The protocol is Version 1.0, May 31, 2020. Recruitment began July 30, 2020, and is anticipated to be completed by October 30, 2020. TRIAL REGISTRATION: This clinical trial has been registered in the Iranian Registry of Clinical Trials (IRCT). The registration number is "IRCT20200506047323N2", https://www.irct.ir/trial/47990 . The registration date is 31 May 2020. FULL PROTOCOL: The full protocol is attached as an additional file, accessible from the Trials website (Additional file 1). In the interest in expediting dissemination of this material, the familiar formatting has been eliminated; this Letter serves as a summary of the key elements of the full protocol.

Identification of bioactive compounds from Glycyrrhiza glabra as possible inhibitor of SARS-CoV-2 spike glycoprotein and non-structural protein-15: a pharmacoinformatics study.

At present, the world is facing a pandemic named as COVID-19, caused by SARS-CoV-2. Traditional Chinese medicine has recommended the use of liquorice (Glycyrrhiza species) in the treatment of infections caused by SARS-CoV-2. Therefore, the present investigation was carried out to identify the active molecule from the liquorice against different protein targets of COVID-19 using an in-silico approach. The molecular docking simulation study of 20 compounds along with two standard antiviral drugs (Lopinavir and Rivabirin) was carried out with the help of Autodock vina software using two protein targets from COVID-19 i.e. spike glycoprotein (PDB ID: 6VSB) and Non-structural Protein-15 (Nsp15) endoribonuclease (PDB ID: 6W01). From the observed binding energy and the binding interactions, glyasperin A showed high affinity towards Nsp15 endoribonuclease with uridine specificity, while glycyrrhizic acid was found to be best suited for the binding pocket of spike glycoprotein and also prohibited the entry of the virus into the host cell. Further, the dynamic behavior of the best-docked molecules inside the spike glycoprotein and Nsp15 endoribonuclease were explored through all-atoms molecular dynamics (MD) simulation study. Several parameters from the MD simulation have substantiated the stability of protein-ligand stability. The binding free energy of both glyasperin A and glycyrrhizic acid was calculated from the entire MD simulation trajectory through the MM-PBSA approach and found to high binding affinity towards the respective protein receptor cavity. Thus, glyasperin A and glycyrrhizic acid could be considered as the best molecule from liquorice, which could find useful against COVID-19. Communicated by Ramaswamy H. Sarma.

Modified Bhramari Pranayama in Covid 19 Infection.

The Coronavirus (2019-Cov-2) infection Covid-19 is highly contagious caused by single stranded RNA virus (+ssRNA) with nucleocapsid and spreading widely all across the world and responsible for more than 3.6 million morbidity and 0.25 million mortality No specific treatment is available till date. The clinical symptoms are mainly upper respiratory leading to diffuse viral pneumonia and multiple organ failure involving. Kidney, Liver and Heart along with coagulopathies. During 2004 (SARS-CoV) pandemic role of nitric oxide in its management is well demonstrated. Nitric Oxide (NO) reversed pulmonary hypertension. Improved severe hypoxia and shortened the stay in ICU and ventilatory support. Nitric Oxide increased the survival rate. The genetic composition of Corona Virus (SARS-CoV) is almost similar to Covid-19, thus indicates good chances of effectiveness or enhancement in results by Nitric Oxide along with other modes in treatment of Covid-19. It has been proved by studies by serendipity humming increases NO Expression dramatically.It is estimated that humming increases the endogenous generation of nitric oxide level by 15-fold. Hypoxia in ARD Syndrome leads to blood coagulation by depression of body defence anticoagulatory and fibrolytic properties along with metabolic acidosis. If we go into hypoxic hypercapnic state no hyper coagulation takes place. Hence Bhramari by enhancing the expression of Nitric Oxide and increased Carbon dioxide by extended exhalation and alkaline pH prevents coagulopathies and morbidity due to Covid-19.