Berberine Inhibits Glioma Cell Migration and Invasion by Suppressing TGF-β1/COL11A1 Pathway and Enhances Chemosensitivity

Background: The expression of collagen type XI alpha 1 chain (COL11A1) is up-regulated in many cancers, affecting the risk of metastasis, drug resistance, and poor prognosis. Berberine is an isoquinoline alkaloid present in many traditional Chinese medicines and it has been shown to reduce collagen accumulation in pulmonary fibrosis, diabetic nephropathy and arthritis. However, its effect on COL11A1 in glioma needs to be further elucidated.Methods: Western blot was performed to detect the expression level of COL11A1 in several glioma cells, and siCOL11A1 was performed to investigate the effect of COL11A1 on the migration and invasion ability of glioma cells. CCK-8, wound healing experiment and transwell experiment were performed to detect the effect of berberine on the proliferation, migration and invasion of glioma cells. The xenografts experiment in nude mice was performed to test the effect of berberine on inhibiting glioma in vivo.Results: Our results showed that COL11A1 is highly expressed in glioma cell lines and associated with migration, invasion, and chemoresistance of glioma cells. Knocking down COL11A1 caused decreased expression of MMPs, Snail, and MGMT. Berberine could inhibit the migration and invasion of glioma cells by suppressing the TGF-β1/COL11A1 pathway and changes actin cytoskeleton arrangement. In addition, berberine increased the chemosensitivity of glioma cells to temozolomide, which may be related to the down-regulation of Snail and MGMT proteins caused by berberine. High-throughput sequencing of xenografts in nude mice also showed that berberine inhibited the expression of COL11A1 in vivo.Conclusions: Our results suggest that berberine that targets COL11A1 to inhibit glioma migration, invasion and chemoresistance, may serve as a promising candidate for the development of anti-glioma drugs in the future.

Lycopene Ameliorates Liver Inflammation and Redox Status in Mice Exposed to Long-Term Cigarette Smoke

Cigarette smoke (CS) is the major cause of preventable death worldwide, and it can also cause damage to extrapulmonary organs, such as the liver, mainly due the generation of reactive oxygen species (ROS). The liver is an essential organ for human survival since it is mainly responsible for the body metabolism and among other things and it is the place where many endogenous and exogenous substances undergo biological transformation. Lycopene is a nonprovitamin A carotenoid found in red fruits and vegetables, and its role as a potent antioxidant is well known. In this study, we hypothesized that lycopene could protect mouse liver against long-term CS exposure. Thirty C57BL/6 mice were exposed to twelve cigarette smoke (12 cigarettes per day) for 60 days and pretreated with 25 mg/kg/day or 50 mg/kg/day of lycopene via orogastric gavage. After euthanasia, the hepatic tissue was collected for histopathological, antioxidant defense, oxidative stress, inflammatory, and collagen deposition analysis. Our analysis demonstrated that lycopene results in a suitable outcome to ameliorate the pathological changes, inflammatory and antioxidant profile in a mouse model of long-term CS exposure, and collagen accumulation in the hepatic extracellular matrix. This study demonstrates for the first time that supplementation of lycopene can be a possible pharmacological tool for the treatment of hepatic damage caused by exposure to long-term CS.

Baicalein Alleviates Fibrosis and Inflammation in Scleroderma Through Regulating B Cell Abnormalities

Background Scleroderma (Systemic sclerosis, SSc) is an autoimmune disorder characterized by multisystem extensive fibrosis, vascular changes and immunological dysregulation. B cell abnormalities play an essential role in the fibrotic pathogenesis of scleroderma by promoting autoantibodies and cytokines release as well as modulating the inflammatory processes. Baicalein, a phenolic flavonoid derived from the Chinese herb Scutellaria baicalensis Georgi, has been used in the treatment of the pathological processes of various fibrotic and inflammatory diseases. Here, we aimed to investigate the effect of baicalein on the major pathological characteristics of SSc both in vitro and in vivo, including fibrosis, B cell abnormalities and inflammation. Methods The effect of baicalein on collagen accumulation and expression of fibrogenic markers in human dermal fibroblasts were analyzed. The antifibrotic features of baicalein and its mechanisms were investigated in the bleomycin (BLM)-induced dermal fibrosis mice model by histologic examination, hydroxyproline assay, enzyme-linked immunosorbent assay and flow cytometry. Results 5-120 µM baicalein significantly abrogated total collagen deposition, decreased soluble collagen secretion, and inhibited the expression of various fibrogenesis molecules, including collagen(COL)1A1, COL1A2, COL3A1, connective tissue growth factor (CTGF), fibronectin, transforming growth factor β1 (TGF-β1) and alpha-smooth muscle actin (α-SMA) in both TGF β1- and PDGF-induced human CCC-ESF-1 dermal fibroblasts. In BLM-induced dermal fibrosis mice model, 25-100 mg/kg baicalein markedly attenuated dermal thickness and collagen accumulation in dose-dependent manners. Baicalein reduced the proportion of B220+ B cells, while increased the percentage of CD27+ memory B cells in the spleen of BLM-induced mice. Consistent with reversing B cell abnormalities, baicalein treatment potently attenuated serum levels of cytokines(IL-1β, IL-2, IL-4, IL6, IL-17A, TNF-α), chemokines (MCP-1, MIP-1β) and autoantibodies (anti-Scl-70, anti-PM-Scl, anti-centromeres and anti-dsDNA). Conclusions Baicalein inhibits TGF-β1 and PDGF- mediated ECM accumulation in human dermal fibroblasts and alleviates the development of experimental dermal fibrosis by reversing B cell abnormalities, reducing autoantibody production and ameliorating inflammation. Baicalein may have the potential to be further developed as a therapeutic candidate against SSc.

Apelin expression deficiency in mice contributes to vascular stiffening by extracellular matrix remodeling of the aortic wall

Numerous recent studies have shown that in the continuum of cardiovascular diseases, the measurement of arterial stiffness has powerful predictive value in cardiovascular risk and mortality and that this value is independent of other conventional risk factors, such as age, cholesterol levels, diabetes, smoking, or average blood pressure. Vascular stiffening is often the main cause of arterial hypertension (AHT), which is common in the presence of obesity. However, the mechanisms leading to vascular stiffening, as well as preventive factors, remain unclear. The aim of the present study was to investigate the consequences of apelin deficiency on the vascular stiffening and wall remodeling of aorta in mice. This factor freed by visceral adipose tissue, is known for its homeostasic role in lipid and vascular metabolisms, or again in inflammation. We compared the level of metabolic markers, inflammation of white adipose tissue (WAT), and aortic wall remodeling from functional and structural approaches in apelin-deficient and wild-type (WT) mice. Apelin-deficient mice were generated by knockout of the apelin gene (APL-KO). From 8 mice by groups, aortic stiffness was analyzed by pulse wave velocity measurements and by characterizations of collagen and elastic fibers. Mann–Whitney statistical test determined the significant data (p < 5%) between groups. The APL-KO mice developed inflammation, which was associated with significant remodeling of visceral WAT, such as neutrophil elastase and cathepsin S expressions. In vitro, cathepsin S activity was detected in conditioned medium prepared from adipose tissue of the APL-KO mice, and cathepsin S activity induced high fragmentations of elastic fiber of wild-type aorta, suggesting that the WAT secretome could play a major role in vascular stiffening. In vivo, remodeling of the extracellular matrix (ECM), such as collagen accumulation and elastolysis, was observed in the aortic walls of the APL-KO mice, with the latter associated with high cathepsin S activity. In addition, pulse wave velocity (PWV) and AHT were increased in the APL-KO mice. The latter could explain aortic wall remodeling in the APL-KO mice. The absence of apelin expression, particularly in WAT, modified the adipocyte secretome and facilitated remodeling of the ECM of the aortic wall. Thus, elastolysis of elastic fibers and collagen accumulation contributed to vascular stiffening and AHT. Therefore, apelin expression could be a major element to preserve vascular homeostasis.

OGR1-dependent regulation of the allergen-induced asthma phenotype

The proton-sensing receptor, ovarian cancer G protein-coupled receptor (OGR1) has been shown to be expressed in airway smooth muscle (ASM) cells and capable of promoting ASM contraction in response to decreased extracellular pH. OGR1 knockout mice (OGR1KO) are reported to be resistant to asthma features induced by inhaled allergen. We recently described certain benzodiazepines as OGR1 activators capable of mediating both pro-contractile and pro-relaxant signaling in ASM cells. Here we assess the effect of treatment with the benzodiazepines lorazepam or sulazepam on the asthma phenotype, in wild type (WT) and OGR1KO mice subjected to inhaled house dust mite (HDM; Dermatophagoides pteronyssius) challenge for three weeks. In contrast to previously published reports, both WT and OGR1KO mice developed significant allergen-induced lung inflammation and airway hyperresponsiveness (AHR). In WT mice, treatment with sulazepam (a Gs-biased OGR1 agonist), but not lorazepam (a balanced OGR1 agonist), prevented allergen-induced AHR, although neither drug inhibited lung inflammation. The protection from development of AHR conferred by sulazepam was absent in OGR1KO mice. Treatment of WT mice with sulazepam also resulted in significant inhibition of HDM-induced collagen accumulation in the lung tissue. These findings suggest OGR1 expression is not a requirement for development of the allergen-induced asthma phenotype, but OGR1 can be targeted by the Gs-biased OGR1 agonist sulazepam (but not the balanced agonist lorazepam) to protect from allergen-induced AHR, possibly mediated via suppression of chronic bronchoconstriction and airway remodeling in the absence of effects on airway inflammation.

Exosomes From ADSCs Attenuate Bleomycin-Induced Skin Fibrosis And Oxidative Stress In Scleroderma Via Circ-Zfyve9 Delivery

Background: Systemic sclerosis (SSc) is autoimmune trait affecting several organs, which is identified by thickening of dermis and connective tissue affected by collagen accumulation, as well as vascular injuries inducing hypoxia. Methods: In this investigation, adipose-derived stem cells (ADSCs) were separated from the ADSC exosomes and a bleomycin-induced SSc mouse model was constructed. We employed high-throughput sequencing to study abnormal expression of circular RNAs (circRNAs) in SSc skin tissues with or without ADSC exosome treatment. The regulatory mechanism and targets were studied using bioinformatics analysis, luciferase reporting analysis, angiogenic differentiation experiments, and RT-qPCR detection. Results: ADSC exosome treatment prevented dermal thickening and fibrosis in bleomycin-induced scleroderma. In addition, circ-Zfyve9 was demonstrated to have an important function in ADSC exosome-mediated skin tissue protection. GPX4 and miR-135 were shown to be circ-Zfyve9 downstream targets. Overexpressing miR-135 or downregulating GPX4 reversed circ-Zfyve9 promotion effects rupon angiopoiesis by promoting lipidosome ROS in EPCs under hypoxic conditions. Overexpressing miR-135 or downregulating GPX4 reversed the circ-Zfyve9 inhibition effect on fibrosis in myofibroblasts under hypoxic conditions. Overexpressing circ-Zfyve9 increased the therapeutic effect of ADSC exosomes. Conclusions: Taken together, the present study results show that the exosomes from ADSCs attenuate bleomycin-induced skin fibrosis and oxidative stress in scleroderma via circ-Zfyve9 delivery.

Aesculetin Inhibits TLR4 and EGFR-Mediated Airway Mucus Hypersecretion and Thickening Induced by Urban Particulate Matter

Objectives uPM10 is microscopic particles of solid or liquid matter suspended in the air. Particulates are the most harmful form of air pollution due to their ability to penetrate deep into the lungs, blood streams and brain, causing diverse health problems. Mucus hypersecretion is a pathological symptom evoked in the bronchi and bronchioles. Excess mucus production can narrow the airways, limit airflow, and accelerate a decline in lung function. Aesculetin, a major component of Sancho tree and Chicory, is known to have antioxidant and anti-inflammatory effects in the vascular and immune system. However, its effect on mucus hypersecretion has been poorly understood. Methods Mice were orally administrated with 10 mg/kg aesculetin and exposed to 6 μg/ml uPM10 for 8 weeks. Airway thickening and collagen accumulation were stained with H&E staining and Masson trichrome staining. The ROS production and mucus hypersecretion were examined with DHE staining, Alcian blue, periodic acid schiff staining and Western blotting. Bronchial epithelial BEAS-2B cells were treated 1–20 μM aesculetin, 20 μg/ml OxPAPC, and 1μM Erlotinib with 2 μg/ml uPM10 for 8 h or 48 h. The protein induction of TLR4, EGFR, and MUC5AC were measured. Results Exposure of mice to uPM10 enhanced total leukocyte number in the BALF with increase in neutrophils and lymphocytes, which was reversed by oral administration of 10 mg/kg aesculetin. In addition, aesculetin attenuated airway thickening along with collagen accumulation, mucus hypersecretion and ROS production elevated by uPM10 inhalation. Nontoxic aesculetin at the concentrations of 1–20 μM inhibited the induction of collagen proteins and MUC5AC in BEAS-2B cells promoted by 2 μg/ml uPM10. On the other hand, uPM10 accelerated the induction of TLR4 and EGFR in BEAS-2B cells, which dampened by 20 μM aesculetin. Both receptors inhibitor reduced the MUC5AC induction in BEAS-2B cells stimulated by uPM10. Conclusions These results demonstrate that aesculetin ameliorated airway thickening and mucus hypersecretion caused by uPM10 inhalation. Therefore, aesculetin maybe a promising agent treating progressive respiratory disorders elicited by urban microparticulates. Funding Sources This work was supported by the BK21 FOUR(Fostering Outstanding Universities for Research, 4220200913807) funded by the National Research Foundation of Korea (NRF).

Imaging Techniques for the Study of Fibrosis in Atrial Fibrillation Ablation: From Molecular Mechanisms to Therapeutical Perspectives

Atrial fibrillation (AF) is the most prevalent form of cardiac arrhythmia. It is often related to diverse pathological conditions affecting the atria and leading to remodeling processes including collagen accumulation, fatty infiltration, and amyloid deposition. All these events generate atrial fibrosis, which contribute to beget AF. In this scenario, cardiac imaging appears as a promising noninvasive tool for monitoring the presence and degree of LA fibrosis and remodeling. The aim of this review is to comprehensively examine the bench mechanisms of atrial fibrosis moving, then to describe the principal imaging techniques that characterize it, such as cardiac magnetic resonance (CMR) and multidetector cardiac computed tomography (MDCT), in order to tailor atrial fibrillation ablation to each individual.