scholarly journals High-content screening of Thai medicinal plants reveals Boesenbergia rotunda extract and its component Panduratin A as anti-SARS-CoV-2 agents

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Phongthon Kanjanasirirat ◽  
Ampa Suksatu ◽  
Suwimon Manopwisedjaroen ◽  
Bamroong Munyoo ◽  
Patoomratana Tuchinda ◽  
...  
Keyword(s):  
Severe Pneumonia ◽  
Inhibitory Effect ◽  
Airway Epithelial Cells ◽  
High Content Screening ◽  
Target Cells ◽  
Airway Epithelial ◽  
Plaque Reduction Assay ◽  
Boesenbergia Rotunda ◽  
First Time ◽  
Panduratin A

AbstractSince December 2019, the emergence of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has caused severe pneumonia, a disease named COVID-19, that became pandemic and created an acute threat to public health. The effective therapeutics are in urgent need. Here, we developed a high-content screening for the antiviral candidates using fluorescence-based SARS-CoV-2 nucleoprotein detection in Vero E6 cells coupled with plaque reduction assay. Among 122 Thai natural products, we found that Boesenbergia rotunda extract and its phytochemical compound, panduratin A, exhibited the potent anti-SARS-CoV-2 activity. Treatment with B. rotunda extract and panduratin A after viral infection drastically suppressed SARS-CoV-2 infectivity in Vero E6 cells with IC50 of 3.62 μg/mL (CC50 = 28.06 µg/mL) and 0.81 μΜ (CC50 = 14.71 µM), respectively. Also, the treatment of panduratin A at the pre-entry phase inhibited SARS-CoV-2 infection with IC50 of 5.30 µM (CC50 = 43.47 µM). Our study demonstrated, for the first time, that panduratin A exerts the inhibitory effect against SARS-CoV-2 infection at both pre-entry and post-infection phases. Apart from Vero E6 cells, treatment with this compound was able to suppress viral infectivity in human airway epithelial cells. This result confirmed the potential of panduratin A as the anti-SARS-CoV-2 agent in the major target cells in human. Since B. rotunda is a culinary herb generally grown in China and Southeast Asia, its extract and the purified panduratin A may serve as the promising candidates for therapeutic purposes with economic advantage during COVID-19 situation.

scholarly journals High-Content Screening of Thai Medicinal Plants Reveals Boesenbergia rotunda Extract and its Component Panduratin A as Anti-SARS-CoV-2 Agents

2020 ◽  
Author(s):  
Phongthon Kanjanasirirat ◽  
Ampa Suksatu ◽  
Suwimon Manopwisedjaroen ◽  
Bamroong Munyoo ◽  
Patoomratana Tuchinda ◽  
...  
Keyword(s):  
Severe Pneumonia ◽  
Inhibitory Effect ◽  
Economic Advantage ◽  
High Content Screening ◽  
Plaque Reduction Assay ◽  
Reduction Assay ◽  
Thai Medicinal Plants ◽  
Boesenbergia Rotunda ◽  
First Time ◽  
Panduratin A

Abstract Since December 2019, the emergence of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has caused severe pneumonia, a disease named COVID-19, that became pandemic and created an acute threat to public health. The effective therapeutics are in urgent need. Here, we developed a high-content screening for the antiviral candidates using fluorescence-based SARS-CoV-2 nucleoprotein detection in Vero E6 cells coupled with plaque reduction assay. Among 122 Thai natural products, we found that Boesenbergia rotunda extract and its phytochemical compound, panduratin A, exhibited the potent anti-SARS-CoV-2 activity. Treatment with B. rotunda extract and panduratin A after viral infection drastically suppressed SARS-CoV-2 infectivity in Vero E6 cells with IC50 of 3.62 μg/mL (CC50 = 28.06 µg/mL) and 0.81 μΜ (CC50=14.71 µM), respectively. Also, the treatment of panduratin A at the pre-entry phase inhibited SARS-CoV-2 infection with IC50 of 5.30 µM (CC50=43.47 µM). Our study demonstrated, for the first time, that panduratin A exerts the inhibitory effect against SARS-CoV-2 infection at both pre-entry and post-infection phases. Since B. rotunda is a culinary herb generally grown in China and Southeast Asia, its extract and the purified panduratin A may serve as the promising candidates for therapeutic purposes with economic advantage during COVID-19 situation.

scholarly journals Infection of bovine well-differentiated airway epithelial cells by Pasteurella multocida: actions and counteractions in the bacteria–host interactions

2020 ◽  
Vol 51 (1) ◽  
Author(s):  
Ang Su ◽  
Jie Tong ◽  
Yuguang Fu ◽  
Sandy Müller ◽  
Yenehiwot Berhanu Weldearegay ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cell Layer ◽  
Airway Epithelial Cells ◽  
Infection Model ◽  
Target Cells ◽  
Respiratory Disorder ◽  
Airway Epithelial ◽  
Epithelial Cell Layer ◽  
Well Differentiated

AbstractPasteurella (P.) multocida is a zoonotic pathogen, which is able to cause respiratory disorder in different hosts. In cattle, P. multocida is an important microorganism involved in the bovine respiratory disease complex (BRDC) with a huge economic impact. We applied air–liquid interface (ALI) cultures of well-differentiated bovine airway epithelial cells to analyze the interaction of P. multocida with its host target cells. The bacterial pathogen grew readily on the ALI cultures. Infection resulted in a substantial loss of ciliated cells. Nevertheless, the epithelial cell layer maintained its barrier function as indicated by the transepithelial electrical resistance and the inability of dextran to get from the apical to the basolateral compartment via the paracellular route. Analysis by confocal immunofluorescence microscopy confirmed the intactness of the epithelial cell layer though it was not as thick as the uninfected control cells. Finally, we chose the bacterial neuraminidase to show that our infection model is a sustainable tool to analyze virulence factors of P. multocida. Furthermore, we provide an explanation, why this microorganism usually is a commensal and becomes pathogenic only in combination with other factors such as co-infecting microorganisms.

scholarly journals Mechanical stretch decreases FAK phosphorylation and reduces cell migration through loss of JIP3-induced JNK phosphorylation in airway epithelial cells

2009 ◽  
Vol 297 (3) ◽  
pp. L520-L529 ◽  
Author(s):  
Leena P. Desai ◽  
Steven R. White ◽  
Christopher M. Waters
Keyword(s):  
Cell Migration ◽  
Epithelial Cells ◽  
Mechanical Strain ◽  
Mechanical Stretch ◽  
Inhibitory Effect ◽  
Airway Epithelial Cells ◽  
Airway Epithelial ◽  
Interacting Protein ◽  
Kinase Activation ◽  
Mapk Cascades

JNK is a nonreceptor kinase involved in the early events that signal cell migration after injury. However, the linkage to early signals required to initiate the migration response to JNK has not been defined in airway epithelial cells, which exist in an environment subjected to cyclic mechanical strain (MS). The present studies demonstrate that the JNK/stress-activated protein kinase-associated protein 1 (JSAP1; also termed JNK-interacting protein 3, JIP3), a scaffold factor for MAPK cascades that links JNK activation to focal adhesion kinase (FAK), are both associated and activated following mechanical injury in 16HBE14o− human airway epithelial cells and that both FAK and JIP3 phosphorylation seen after injury are decreased in cells subjected to cyclic MS. Overexpression of either wild-type (WT)-FAK or WT-JIP3 enhanced phosphorylation and kinase activation of JNK and reduced the inhibitory effect of cyclic MS. These results suggest that cyclic MS impairs signaling of cell migration after injury via a pathway that involves FAK-JIP3-JNK.

A system to impose prescribed homogenous strains on cultured cells

2001 ◽  
Vol 91 (4) ◽  
pp. 1600-1610 ◽  
Author(s):  
Christopher M. Waters ◽  
Matthew R. Glucksberg ◽  
Eugene P. Lautenschlager ◽  
Chyh-Woei Lee ◽  
Reed M. Van Matre ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Cultured Cells ◽  
Large Strain ◽  
Mechanical Strain ◽  
Airway Epithelial Cells ◽  
Airway Epithelial ◽  
Degree Of Anisotropy ◽  
High Degree ◽  
Membrane Shape ◽  
First Time

There is presently significant interest in cellular responses to physical forces, and numerous devices have been developed to apply stretch to cultured cells. Many of the early devices were limited by the heterogeneity of deformation of cells in different locations and by the high degree of anisotropy at a particular location. We have therefore developed a system to impose cyclic, large-strain, homogeneous stretch on a multiwell surface-treated silicone elastomer substrate plated with pulmonary epithelial cells. The pneumatically driven mechanism consists of four plates each with a clamp to fix one edge of the cruciform elastomer substrate. Four linear bearings set at predetermined angles between the plates ensure a constant ratio of principal strains throughout the stretch cycle. We present the design of the device and membrane shape, the surface modifications of the membrane to promote cell adhesion, predicted and experimental measurements of the strain field, and new data using cultured airway epithelial cells. We present for the first time the relationship between the magnitude of cyclic mechanical strain and the extent of wound closure and cell spreading.

scholarly journals Ets-1 is a negative regulator of Th17 differentiation

2007 ◽  
Vol 204 (12) ◽  
pp. 2825-2835 ◽  
Author(s):  
Jacques Moisan ◽  
Roland Grenningloh ◽  
Estelle Bettelli ◽  
Mohamed Oukka ◽  
I-Cheng Ho
Keyword(s):  
Th17 Cells ◽  
Inhibitory Effect ◽  
Airway Epithelial Cells ◽  
Negative Regulator ◽  
Dependent Manner ◽  
Airway Epithelial ◽  
Mucus Production ◽  
Th Cells ◽  
Th17 Differentiation ◽  
High Level

IL-17 is a proinflammatory cytokine that plays a role in the clearance of extracellular bacteria and contributes to the pathology of many autoimmune and allergic conditions. IL-17 is produced mainly by a newly characterized subset of T helper (Th) cells termed Th17. Although the role of Th17 cells in the pathology of autoimmune diseases is well established, the transcription factors regulating the differentiation of Th17 cells remain poorly characterized. We report that Ets-1–deficient Th cells differentiated more efficiently to Th17 cells than wild-type cells. This was attributed to both low IL-2 production and increased resistance to the inhibitory effect of IL-2 on Th17 differentiation. The resistance to IL-2 suppression was caused by a defect downstream of STAT5 phosphorylation, but was not caused by a difference in the level of RORγt. Furthermore, Ets-1–deficient mice contained an abnormally high level of IL-17 transcripts in their lungs and exhibited increased mucus production by airway epithelial cells in an IL-17–dependent manner. Based on these observations, we report that Ets-1 is a negative regulator of Th17 differentiation.

scholarly journals Extracellular Vesicle-Mediated siRNA Delivery, Protein Delivery, and CFTR Complementation in Well-Differentiated Human Airway Epithelial Cells

Genes ◽  
2020 ◽  
Vol 11 (4) ◽  
pp. 351 ◽  
Author(s):  
Brajesh K. Singh ◽  
Ashley L. Cooney ◽  
Sateesh Krishnamurthy ◽  
Patrick L. Sinn
Keyword(s):  
Epithelial Cells ◽  
Airway Epithelial Cells ◽  
Target Cells ◽  
Airway Epithelial ◽  
Long Distance ◽  
Human Airway ◽  
Well Differentiated ◽  
Human Airway Epithelial Cells

Extracellular vesicles (EVs) are a class of naturally occurring secreted cellular bodies that are involved in long distance cell-to-cell communication. Proteins, lipids, mRNA, and miRNA can be packaged into these vesicles and released from the cell. This information is then delivered to target cells. Since EVs are naturally adapted molecular messengers, they have emerged as an innovative, inexpensive, and robust method to deliver therapeutic cargo in vitro and in vivo. Well-differentiated primary cultures of human airway epithelial cells (HAE) are refractory to standard transfection techniques. Indeed, common strategies used to overexpress or knockdown gene expression in immortalized cell lines simply have no detectable effect in HAE. Here we use EVs to efficiently deliver siRNA or protein to HAE. Furthermore, EVs can deliver CFTR protein to cystic fibrosis donor cells and functionally correct the Cl− channel defect in vitro. EV-mediated delivery of siRNA or proteins to HAE provides a powerful genetic tool in a model system that closely recapitulates the in vivo airways.

β-Casein(94-123)-derived peptides differently modulate production of mucins in intestinal goblet cells

2014 ◽  
Vol 82 (1) ◽  
pp. 36-46 ◽  
Author(s):  
Pascale Plaisancié ◽  
Rachel Boutrou ◽  
Monique Estienne ◽  
Gwénaële Henry ◽  
Julien Jardin ◽  
...  
Keyword(s):  
Bioactive Peptides ◽  
Opioid Peptide ◽  
Inhibitory Effect ◽  
Airway Epithelial Cells ◽  
Amino Acid Residues ◽  
Airway Epithelial ◽  
Inhibitory Peptide ◽  
Intestinal Mucus ◽  
Muc4 Expression

We recently reported the identification of a peptide from yoghurts with promising potential for intestinal health: the sequence (94-123) of bovine β-casein. This peptide, composed of 30 amino acid residues, maintains intestinal homoeostasis through production of the secreted mucin MUC2 and of the transmembrane-associated mucin MUC4. Our study aimed to search for the minimal sequence responsible for the biological activity of β-CN(94-123) by using several strategies based on (i) known bioactive peptides encrypted in β-CN(94-123), (ii) in silico prediction of peptides reactivity and (iii) digestion of β-CN(94-123) by enzymes of intestinal brush border membranes. The revealed sequences were tested in vitro on human intestinal mucus-producing HT29-MTX cells. We demonstrated that β-CN(108-113) (an ACE-inhibitory peptide) and β-CN(114-119) (an opioid peptide named neocasomorphin-6) up-regulated MUC4 expression whereas levels of the secreted mucins MUC2 and MUC5AC remained unchanged. The digestion of β-CN(94-123) by intestinal enzymes showed that the peptides β-CN(94-108) and β-CN(117-123) were present throughout 1·5 to 3 h of digestion, respectively. These two peptides raised MUC5AC expression while β-CN(117-123) also induced a decrease in the level of MUC2 mRNA and protein. In addition, this inhibitory effect was reproduced in airway epithelial cells. In conclusion, β-CN(94-123) is a multifunctional molecule but only the sequence of 30 amino acids has a stimulating effect on the production of MUC2, a crucial factor of intestinal protection.

scholarly journals Hydrogen Sulfide Inhibits TMPRSS2 in Human Airway Epithelial Cells: Implications for SARS-CoV-2 Infection

Biomedicines ◽  
2021 ◽  
Vol 9 (9) ◽  
pp. 1273
Author(s):  
Giulia Pozzi ◽  
Elena Masselli ◽  
Giuliana Gobbi ◽  
Prisco Mirandola ◽  
Luis Taborda-Barata ◽  
...  
Keyword(s):  
Hydrogen Sulfide ◽  
Epithelial Cells ◽  
Airway Epithelial Cells ◽  
Surface Proteins ◽  
Lower Airway ◽  
Target Cells ◽  
Airway Epithelial ◽  
Respiratory Epithelial Cells ◽  
Respiratory Epithelial

The COVID-19 pandemic has now affected around 190 million people worldwide, accounting for more than 4 million confirmed deaths. Besides ongoing global vaccination, finding protective and therapeutic strategies is an urgent clinical need. SARS-CoV-2 mostly infects the host organism via the respiratory system, requiring angiotensin-converting enzyme 2 (ACE2) and transmembrane protease serine 2 (TMPRSS2) to enter target cells. Therefore, these surface proteins are considered potential druggable targets. Hydrogen sulfide (H2S) is a gasotransmitter produced by several cell types and is also part of natural compounds, such as sulfurous waters that are often inhaled as low-intensity therapy and prevention in different respiratory conditions. H2S is a potent biological mediator, with anti-oxidant, anti-inflammatory, and, as more recently shown, also anti-viral activities. Considering that respiratory epithelial cells can be directly exposed to H2S by inhalation, here we tested the in vitro effects of H2S-donors on TMPRSS2 and ACE2 expression in human upper and lower airway epithelial cells. We showed that H2S significantly reduces the expression of TMPRSS2 without modifying ACE2 expression both in respiratory cell lines and primary human upper and lower airway epithelial cells. Results suggest that inhalational exposure of respiratory epithelial cells to natural H2S sources may hinder SARS-CoV-2 entry into airway epithelial cells and, consequently, potentially prevent the virus from spreading into the lower respiratory tract and the lung.

Negative feedback between secretory and cytosolic phospholipase A2 and their opposing roles in ovalbumin-induced bronchoconstriction in rats

2005 ◽  
Vol 288 (3) ◽  
pp. L523-L529 ◽  
Author(s):  
Sarit Offer ◽  
Saul Yedgar ◽  
Ouri Schwob ◽  
Miron Krimsky ◽  
Haim Bibi ◽  
...  
Keyword(s):  
Negative Feedback ◽  
Inflammatory Cells ◽  
Airway Epithelial Cells ◽  
Airway Epithelial ◽  
Membrane Phospholipids ◽  
Cytosolic Phospholipase ◽  
Inflammatory Processes ◽  
A Cell ◽  
First Time

Phospholipase A2 (PLA2) hydrolyzes cell membrane phospholipids (PL) to produce arachidonic acid and lyso-PL. The PLA2 enzymes include the secretory (sPLA2) and cytosolic (cPLA2) isoforms, which are assumed to act synergistically in production of eicosanoids that are involved in inflammatory processes. However, growing evidence raises the possibility that in airways and asthma-related inflammatory cells (eosinophils, basophils), the production of the bronchoconstrictor cysteinyl leukotrienes (CysLT) is linked exclusively to sPLA2, whereas the bronchodilator prostaglandin PGE2 is produced by cPLA2. It has been further reported that the capacity of airway epithelial cells to produce CysLT is inversely proportional to PGE2 production. This seems to suggest that sPLA2 and cPLA2 play opposing roles in asthma pathophysiology and the possibility of a negative feedback between the two isoenzymes. To test this hypothesis, we examined the effect of a cell-impermeable extracellular sPLA2 inhibitor on bronchoconstriction and PLA2 expression in rats with ovalbumin (OVA)-induced asthma. It was found that OVA-induced bronchoconstriction was associated with elevation of lung sPLA2 expression and CysLT production, concomitantly with suppression of cPLA2 expression and PGE2 production. These were reversed by treatment with the sPLA2 inhibitor, resulting in amelioration of bronchoconstriction and reduced CysLT production and sPLA2 expression, concomitantly with enhanced PGE2 production and cPLA2 expression. This study demonstrates, for the first time in vivo, a negative feedback between sPLA2 and cPLA2 and assigns opposing roles for these enzymes in asthma pathophysiology: sPLA2 activation induces production of the bronchoconstrictor CysLT and suppresses cPLA2 expression and the subsequent production of the bronchodilator PGE2.

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