scholarly journals Antifibrotic effect of Ocimum basilicum L. and linalool on arecoline-induced fibrosis in human buccal fibroblasts

2018 ◽  
Vol 3 ◽  
pp. 2057178X1876447 ◽  
Author(s):  
Pooja Adtani ◽  
Narasimhan Malathi ◽  
Kannan Ranganathan ◽  
Sivaswamy Lokeswari ◽  
Alan Mathew Punnoose
Keyword(s):  
Transforming Growth Factor Beta ◽  
In Vitro Model ◽  
Transforming Growth Factor ◽  
Oral Submucous Fibrosis ◽  
Ocimum Basilicum ◽  
Morphological Alterations ◽  
Antifibrotic Effect ◽  
Submucous Fibrosis ◽  
Vitro Model

Aim: To explore Ocimum basilicum L. (sweet basil) and linalool for their antifibrotic activity in an arecoline-induced in vitro fibrotic model. Methods: Leaf extract of O. basilicum L. (LEOB) and linalool were used as experimental agents to test their antifibrogenic activity in vitro. Half-maximal inhibitory concentration (IC50) for arecoline, ethanolic LEOB, and linalool was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. To evaluate the antifibrotic effect of ethanolic LEOB and linalool on pretreatment, that is, both the testing agents were added to the human buccal fibroblasts (HBFs) prior to induction with arecoline, and reverse transcriptase polymerase chain reaction (RT-PCR) was carried out to study the response of transforming growth factor beta (TGFβ), collagen 1 subtype A2 (COL1A2), and collagen 3 subtype A1 (COL3A1). To appreciate the morphological alterations in HBFs on treatment with arecoline, ethanolic LEOB, and linalool, Masson’s trichrome staining was performed. Results: Arecoline enhanced fibrotic activity by upregulating TGFβ1, COL1A2, and COL3A1 levels, whereas ethanolic LEOB and linalool on pretreatment significantly downregulated the increased levels of TGFβ1, COL1A2, and COL3A1 in primary HBF cell cultures. Conclusion and implication to clinic: Both ethanolic LEOB and linalool exhibited significant antifibrotic activity in an in vitro model. Further studies in an in vitro model can help attain a foundation for an herbal formulation in gel form that can be prescribed to patients diagnosed with oral submucous fibrosis for topical application. It can also be used synergistically with Western medicine.

scholarly journals Long-Term Maintenance of Viable Adult Rat Sertoli Cells Able to Establish Testis Barrier Components and Function in Response to Androgens

Cells ◽  
2021 ◽  
Vol 10 (9) ◽  
pp. 2405
Author(s):  
Hassan Kabbesh ◽  
Muhammad A. Riaz ◽  
Alexandra D. Jensen ◽  
Georgios Scheiner-Bobis ◽  
Lutz Konrad
Keyword(s):  
Tight Junctions ◽  
Transforming Growth Factor Beta ◽  
Sertoli Cells ◽  
In Vitro Model ◽  
Transforming Growth Factor ◽  
Bone Morphogenetic Protein 2 ◽  
Adult Rat ◽  
Vitro Model

A protocol for the isolation and long-term propagation of adult rat Sertoli cells (SCs) using conditional reprogramming (CR) was developed and the formation of tight junctions as an in vitro model for the blood testis barrier (BTB) was studied. Three pure primary SC lines were isolated successfully and maintained for several months without significant changes in expression levels of SC-typical markers such as SRY-box transcription factor 9 (SOX9), transferrin, clusterin, androgen receptor (AR), and GATA binding protein 1 (GATA1). In addition to AR expression, the tight junction proteins, zonula occludens-1 (ZO-1) and the junctional adhesion molecule-3 (JAM-3), were upregulated and the SC barrier integrity was enhanced by testosterone. Peritubular/myoid cells did not increase the tightness of the SC. The cytokines, interleukin-6 (IL-6), bone morphogenetic protein-2 (BMP2), and transforming growth factor beta-3 (TGF-β3), negatively affected the tightness of the SC barrier. We have established a protocol for the isolation and long-term propagation of highly pure primary adult rat SCs, which are able to respond to androgen treatments, to form tight junctions and to maintain the mRNA expression of SC-specific genes. By applying this new method, adult SCs can now be analyzed in more detail and might serve as an in vitro model for the study of many SC functions.

scholarly journals Proposal for experimental in vitro model to assess morphological alterations in erythrocytes exposed to 5.25% NaOCl

2016 ◽  
Vol 20 (4) ◽  
pp. e241-e245
Author(s):  
Roberto Arroyo Cervantes ◽  
Sergio Iván Cuin Macedo ◽  
Benigno Miguel Calderón Rojas ◽  
Diana Ened Rodríguez Zaragoza ◽  
Héctor Ruiz Reyes
Keyword(s):  
In Vitro Model ◽  
Morphological Alterations ◽  
Vitro Model

scholarly journals Metformin activity in an in vitro model of posterior capsule opacification

2018 ◽  
Vol 17 (4) ◽  
pp. 105-112
Author(s):  
Jade Marie Lasiste ◽  
Pablo Zoroquiain ◽  
Denise Miyamoto ◽  
Miguel Burnier
Keyword(s):  
Growth Factor ◽  
Western Blot ◽  
In Vitro Model ◽  
Transforming Growth Factor ◽  
Posterior Capsule Opacification ◽  
Posterior Capsule ◽  
Vitro Model ◽  
And Migration ◽  
E Cadherin

Purpose: To determine the activity of metformin in an in vitro model of posterior capsule opacification (PCO).Design: Experimental laboratory research. Methods: The HLE-B3 lens epithelial cell line was treated with PCO induction media (PCOM) supplemented with transforming growth factor-beta (TGF-β) and fibroblast growth factor (FGF). Different metformin concentrations (0-100 mM) were used. The following cellular parameters were assessed: (1) survival, using a viability assay; (2) morphology, via microscopy and image analysis; (3) migration, using the wound assay; (4) and expression of epithelial (Pax6, E-cadherin) and mesenchymal (α-smooth muscle actin or α-SMA, fibronectin) markers via Western blot. Expression of the uptake receptor SLC22A1 was evaluated in HLE-B3 and in human donor eyes with Western blot and immunohistochemistry, respectively. Statistical analysis of variance (ANOVA) with Tukey post-hoc test was done for analysis of cytotoxicity, morphology and migration data. Results: Metformin was lethal to half (LC50) of the cells at 30 mM, and a decrease in viability (P<0.05) was noted at 5 mM. LECs in PCOM treated with 1 mM metformin showed increased Pax6 and E-cadherin and decreased α-SMA and fibronectin expression. LECs in PCOM treated with metformin also maintained epithelial morphology. Migration was inhibited with 0.5 mM metformin (P<0.05). Both HLE-B3 and the lens epithelium in donor eyes were found to express SLC22A1.Conclusion: Metformin decreased survival and migration in LECs, maintaining epithelial phenotype and reducing mesenchymal marker expression. Metformin therefore has potential as an adjunct in PCO prevention.Financial Disclosures: This work was partially funded by Mitacs Canada.

scholarly journals Small Molecule “Silmitasertib” Repurposed as Inhibitor of Transforming Growth Factor Beta 1 for the Development of Therapeutics for Oral Submucous Fibrosis

2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Nezar Boreak
Keyword(s):  
Growth Factor ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Oral Submucous Fibrosis ◽  
Normal Tissues ◽  
Premalignant Condition ◽  
Submucous Fibrosis ◽  
Transformation Potential ◽  
Growth Factor Beta ◽  
Tgf Β1

Oral submucous fibrosis (OSMF) is considered a premalignant condition characterized by aggressive fibrosis of the submucosal tissues of the oral cavity reflecting its malignant transformation potential. Activation of transforming growth factor beta (TGF-β) signaling has been reported to lead increased collagen production and fibrosis. Recently, significant upregulation of TGF-β1 has been reported in OSMF as compared to normal tissues. Therefore, inhibition of the TGF-β1 may pave for the development of therapeutics of OSMF. Based on the structure-assisted drug designing, we found “silmitasertib” as potent inhibitor of TGF-β1. We suggest that this molecule can be validated and implemented for the treatment of OSMF.

scholarly journals Kasugamycin is a novel chitinase 1 inhibitor with strong antifibrotic effects on pulmonary fibrosis

2021 ◽  
Author(s):  
Jae-Hyun Lee ◽  
Chang-Min Lee ◽  
Mun-Ock Kim ◽  
Jin Wook Park ◽  
Suchitra Kamle ◽  
...  
Keyword(s):  
Growth Factor ◽  
Pulmonary Fibrosis ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Glycosyl Hydrolase ◽  
Aminoglycoside Antibiotic ◽  
Therapeutic Drug ◽  
Antifibrotic Effect

Rationale: Pulmonary fibrosis is a devastating lung disease with few therapeutic options. Chitinase 1 (CHIT1), an 18 glycosyl hydrolase family member, contributes to the pathogenesis of pulmonary fibrosis through regulation of Transforming Growth Factor (TGF)-β signaling and effector function. Therefore, CHIT1 is a potential therapeutic target of pulmonary fibrosis. Objectives: This study aimed to identify and characterize a druggable CHIT1 inhibitor with strong antifibrotic activity and minimal toxicity for therapeutic application to pulmonary fibrosis. Methods: Extensive screening of small molecule libraries identified the aminoglycoside antibiotic Kasugamycin as a potent CHIT1 inhibitor. Measurements and Main Results: Elevated levels of CHIT1 were detected in the lungs of patients with pulmonary fibrosis. In vivo bleomycin- and TGF-β-stimulated murine models of pulmonary fibrosis, Kasugamycin showed impressive anti-fibrotic effects in both preventive and therapeutic conditions. In vitro studies also demonstrated that Kasugamycin inhibits fibrotic macrophage activation, fibroblast proliferation and myofibroblast transformation. Null mutation of transforming growth factor beta associated protein 1 (TGFBRAP1), a recently identified CHIT1 interacting signaling molecule, phenocopied antifibrotic effects of Kasugamycin in in vivo lungs and in vitro fibroblasts responses. Kasugamycin inhibits physical association between CHIT1 and TGFBRAP1 with decreased levels of SMAD4 association, suggesting that antifibrotic effect of Kasugamycin is mediated through regulation of TGFBRAP1, at least in part. Conclusions: These studies demonstrate that Kasugamycin is a novel CHIT1 inhibitor with strong antifibrotic effect that can be further developed as an effective and safe therapeutic drug for pulmonary fibrosis.

Influence of cytokines and growth factors in ANG II-mediated collagen upregulation by fibroblasts in rats: role of myocytes

2004 ◽  
Vol 287 (1) ◽  
pp. H107-H117 ◽  
Author(s):  
Sagartirtha Sarkar ◽  
Elangovan Vellaichamy ◽  
David Young ◽  
Subha Sen
Keyword(s):  
In Vitro Model ◽  
Transforming Growth Factor ◽  
Cardiac Fibroblasts ◽  
Ang Ii ◽  
Tnf Α ◽  
Paracrine Action ◽  
Vitro Model ◽  
Necrosis Factor

Abnormal stiffness and altered cardiac function arising from abnormal collagen deposition occur in hypertrophy and heart failure. ANG II has been shown to play a role in this process. To evaluate the mechanism, we developed an in vitro model by subjecting fibroblasts to ANG II treatment in the presence or absence of myocytes in coculture ( 25 ). Employing this model, we demonstrated that ANG II-induced collagen gene transcription in cardiac fibroblasts was potentiated by myocyte-derived factors. In attempting to identify mechanisms of collagen upregulation and to define the role of myocytes, we found that interleukin (IL)-6, tumor necrosis factor (TNF)-α, and the transforming growth factor (TGF)-β superfamily were also involved in collagen upregulation. Collagen transcripts were increased after fibroblasts were treated with IL-6 (20–50 ng/ml) and TNF-α (0.1–0.5 ng/ml). In this study, we show that cardiomyocytes induce secretion of active TGF-β in the presence of ANG II and that a paracrine action of TGF-β subsequently induces different cytokines (IL-6) in fibroblasts, thereby promoting collagen synthesis. The cross-talk between myocytes and fibroblasts and involvement of these cytokines in the upregulation of collagen transcript levels are novel findings that may explain their possible roles in the upregulation of collagen.

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