Correlation matrixes representing the Pearson's correlation coefficient between pairs of data on mRNA expression in intestinal explants. Murine intestinal fibrosis was induced by the heterotopic transplant of colonic tissue in mice receiving a daily intraperitoneal injection of the autophagy stimulator rapamycin (1.25 mg/kg mice), the autophagy inhibitor 3-methyladenine (3-MA, 10 mg/kg mice) or their vehicle (DMSO 1%, n = 5 each). The intestinal grafts resected seven days after transplantation were analyzed with regard to their mRNA expression of genes involved in i) fibrosis Correlation matrixes representing the Pearson's correlation coefficient between pairs of data on mRNA expression in intestinal explants. Murine intestinal fibrosis was induced by the heterotopic transplant of colonic tissue in mice receiving a daily intraperitoneal injection of the autophagy stimulator rapamycin (1.25 mg/kg mice), the autophagy inhibitor 3-methyladenine (3-MA, 10 mg/kg mice) or their vehicle (DMSO 1%, n = 5 each). The intestinal grafts resected seven days after transplantation were analyzed with regard to their mRNA expression of genes involved in (i) fibrosis and epithelial to mesenchymal transition; and (ii) inflammation: the general macrophage marker (F4/80), pro-inflammatory (red), anti-inflammatory (blue) and pro-fibrotic (purple) molecules. The data were organized taking as references: (i) the fibroblast marker vimentin (in all groups); and (ii) the macrophage marker which correlates with a higher number of markers of inflammation in each experimental group (vehicle: CD163; rapamycin: F4/80, 3-MA: CD16). The pairwise comparison heatmaps were performed using the online resource available at http://www.heatmapper.ca, and interpreted according to its authors' instructions [7].

Correlation matrixes representing the Pearson's correlation coefficient between pairs of data on mRNA expression in intestinal explants. Murine intestinal fibrosis was induced by the heterotopic transplant of colonic tissue in mice receiving a daily intraperitoneal injection of the autophagy stimulator rapamycin (1.25 mg/kg mice), the autophagy inhibitor 3-methyladenine (3-MA, 10 mg/kg mice) or their vehicle (DMSO 1%, n = 5 each). The intestinal grafts resected seven days after transplantation were analyzed with regard to their mRNA expression of genes involved in i) fibrosis Correlation matrixes representing the Pearson's correlation coefficient between pairs of data on mRNA expression in intestinal explants. Murine intestinal fibrosis was induced by the heterotopic transplant of colonic tissue in mice receiving a daily intraperitoneal injection of the autophagy stimulator rapamycin (1.25 mg/kg mice), the autophagy inhibitor 3-methyladenine (3-MA, 10 mg/kg mice) or their vehicle (DMSO 1%, n = 5 each). The intestinal grafts resected seven days after transplantation were analyzed with regard to their mRNA expression of genes involved in (i) fibrosis and epithelial to mesenchymal transition; and (ii) inflammation: the general macrophage marker (F4/80), pro-inflammatory (red), anti-inflammatory (blue) and pro-fibrotic (purple) molecules. The data were organized taking as references: (i) the fibroblast marker vimentin (in all groups); and (ii) the macrophage marker which correlates with a higher number of markers of inflammation in each experimental group (vehicle: CD163; rapamycin: F4/80, 3-MA: CD16). The pairwise comparison heatmaps were performed using the online resource available at http://www.heatmapper.ca, and interpreted according to its authors' instructions [7].

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We recently observed reduced autophagy in Crohn’s disease patients and an anti-inflammatory effect of autophagy stimulation in murine colitis, but both anti- and pro-fibrotic effects are associated with autophagy stimulation in different tissues, and fibrosis is a frequent complication of Crohn’s disease. Thus, we analyzed the effects of pharmacolo...

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... In a murine model of intestinal fibrosis, it has been demonstrated that autophagy inhibition aggravates, while autophagy stimulation prevents, fibrosis. 39 In addition to the 'in vitro' observation that fibroblast autophagy, and its pharmacological regulation, affects collagen degradation, the tissue inflammatory response that accompanied fibrosis was significantly affected by autophagy modulation. In particular, in mice treated with the autophagy inhibitor, an increased expression of pro-inflammatory mediators and pro-fibrogenic 40 CD16 + -M2 macrophage infiltration was observed, while stimulation of autophagy with rapamycin increased the expression in mice of anti-inflammatory mediators and infiltration of macrophages with a regulatory/anti-inflammatory profile. ...
... In particular, in mice treated with the autophagy inhibitor, an increased expression of pro-inflammatory mediators and pro-fibrogenic 40 CD16 + -M2 macrophage infiltration was observed, while stimulation of autophagy with rapamycin increased the expression in mice of anti-inflammatory mediators and infiltration of macrophages with a regulatory/anti-inflammatory profile. 39 Thus, autophagy, macrophages and related cytokine modulation seem to play a relevant role in the fibrogenic process. This view is reinforced by the observations made in the murine model of TNBS chronic colitis. ...
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Background and Aims Intestinal fibrosis is a common complication of inflammatory bowel diseases. Medical treatment of intestinal fibrosis is an unmet therapeutic need. CD147 overexpression can induce myofibroblast differentiation associated with extracellular matrix deposition, favouring the development of fibrosis. To understand whether CD147 may promote intestinal fibrosis, we analysed its expression and blocked its function by using its specific inhibitor AC-73 [3-{2-[([1,1’-biphenyl]-4-ylmethyl) amino]-1-hydroxyethyl} phenol] in the murine TNBS [trinitrobenzenesulfonic acid]-chronic colitis model associated with intestinal fibrosis. Methods TNBS chronic colitis was induced by weekly intrarectal administration of escalating doses of TNBS. Ethanol-treated and untreated mice were used as controls. Separated groups of TNBS, ethanol-treated or untreated mice received AC-73 or vehicle administered intraperitoneally from day 21 to day 49. At day 49, mice were killed, and colons collected for histological analysis, protein and RNA extraction. CD147, α-SMA and activated TGF-β1 protein levels, CD147/ERK/STAT3 signalling pathway and autophagy were assessed by Western blot, collagen and inflammatory/fibrogenic cytokines mRNA tissue content by quantitative PCR. Results In mice with chronic TNBS colitis, CD147 protein level increased during fibrosis development in colonic tissue, as compared to control mice. CD147 inhibition by AC-73 treatment reduced intestinal fibrosis, collagen and cytokine mRNA tissue content, without significant modulation of activated TGF-β1 protein tissue content. AC-73 inhibited CD147/ERK1/2 and STAT3 signalling pathway activation and induced autophagy. Conclusions CD147 is a potential new target for controlling intestinal fibrosis and its inhibitor, AC-73, might represent a potential new anti-fibrotic therapeutic option in IBD.
... Total RNA from cells and murine and human tissues was isolated using direct-zol RNA MiniPrep Plus R2072 from ZymoResearch according to the manufacturer's instructions. Mice tissue and intestinal resections were homogenated with TRI Reagent ® (ZymoResearch, Irvine, CA, USA), using the GentleMACS Dissociator (Milteny Biotech, Gladbach, Germany) as previously described [36]. cDNA was obtained from previously isolated RNA by reverse transcription PCRusing the the PrimeScript RT reagent Kit (Takara Bitechnology, Dalian, China). ...
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Intestinal epithelial cells (IECs) constitute a defensive physical barrier in mucosal tissues and their disruption is involved in the etiopathogenesis of several inflammatory pathologies, such as Ulcerative Colitis (UC). Recently, the succinate receptor SUCNR1 was associated with the activation of inflammatory pathways in several cell types, but little is known about its role in IECs. We aimed to analyze the role of SUCNR1 in the inflammasome priming and its relevance in UC. Inflammatory and inflammasome markers and SUCNR1 were analyzed in HT29 cells treated with succinate and/or an inflammatory cocktail and transfected with SUCNR1 siRNA in a murine DSS model, and in intestinal resections from 15 UC and non-IBD patients. Results showed that this receptor mediated the inflammasome, priming both in vitro in HT29 cells and in vivo in a murine chronic DSS-colitis model. Moreover, SUNCR1 was also found to be involved in the activation of the inflammatory pathways NFкB and ERK pathways, even in basal conditions, since the transient knock-down of this receptor significantly reduced the constitutive levels of pERK-1/2 and pNFкB and impaired LPS-induced inflammation. Finally, UC patients showed a significant increase in the expression of SUCNR1 and several inflammasome components which correlated positively and significantly. Therefore, our results demonstrated a role for SUCNR1 in basal and stimulated inflammatory pathways in intestinal epithelial cells and suggested a pivotal role for this receptor in inflammasome activation in UC.
... Our study also confirmed that proinflammatory markers were not correlated with fibrosis-associated markers, and preventive intervention did not achieve better anti-fibrotic efficacy. Defects in autophagy have been linked with fibrosis in different organs, especially in the intestine, and several mutated autophagy genes are associated with the development of structuring and penetrating complications in IBD [33,34]. Of note, the interaction between autophagy and EMT has been reported in other studies, with a recent study showing that autophagy inhibition induces EMT via the p62/SQSMT1-NFκB-Snail2 pathway in alveolar epithelial cells and promotes local myofibroblast differentiation through paracrine signals [23]. ...
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... Other studies observed that autophagy seems to inhibit intestinal fibrosis by modulating the function of the innate immune system and the mesenchymal activity. [3,12] (7) Balance intestinal epithelial cells (ISCs) regeneration. The critical role of autophagy in maintaining ISC functions under different physiological conditions has been discovered only in recent years. ...
... Autophagy, a 'self-digestion' process that aims to reuse cellular components to adapt to starvation and stress, participates in many aspects of biology [29]. Indeed, studies have demonstrated that autophagy is a potential target in preventing fibrosis [28,30]. Fibroblast proliferation and migration play a crucial role in arthrofibrosis. ...
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Post-surgery arthrofibrosis is one of the most restrictive factors in the development of intra-articular surgery and has presented tremendous obstacles for most orthopaedic surgeons. Tanshinone IIA (Tan IIA), a key active ingredient of Den-shen, has been used to treat fibrosis-related diseases, such as pulmonary, hepatic and myocardial fibrosis. In the present study, we aimed to investigate the effects of Tan IIA on post-surgery arthrofibrosis in vivo and in vitro. Histological analysis indicated that topical application of Tan IIA (10 mg/mL) could significantly alleviate postsurgery arthrofibrosis in rabbits. Immunohistochemistry results showed that proliferating cell nuclear antigen (PCNA) and tubulin protein expression was inhibited, whereas LC3 was activated in vivo. In vitro, EdU and flow cytometry assays demonstrated that Tan IIA could inhibit fibroblast proliferation by arresting cells in G2 phase. Scratch, Transwell and cytoskeleton protein immunofluorescence assays revealed that fibroblast migration was attenuated. Interestingly, LC3 immunofluorescence staining and transmission electron microscopy indicated that autophagy flux could be induced in fibroblasts by Tan IIA. However, the inhibitory effects of Tan IIA against fibroblast proliferation and migration were partially restored when fibroblast autophagy was suppressed after combined treatment with the autophagy inhibitor 3-methyladenine (3-MA). Finally, the expression of p-mTOR was suppressed in a dose-dependent manner after Tan IIA treatment but partially restored when Tan IIA treatment was combined with 3-MA intervention. The inhibitory effect of Tan IIA against fibroblast proliferation and migration may be related to autophagy induction mediated by the PI3K and AMPK-mTOR signaling pathway.
... Ye et al. revealed that ursodeoxycholic acid displayed anti-fibrotic role in hepatic stellate cells involving autophagy inhibition, and rapamycin was found to impair its treatment effect [12]. But there are some different views that the progression of several fibrotic diseases can be alleviated by activation of autophagy, such as intestinal fibrosis [13,14], myocardial fibrosis [15], peritoneal fibrosis [16], and idiopathic pulmonary fibrosis [17][18][19]. However, the role of autophagy in the process of conjunctival fibrosis has not been studied. ...
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New findings: What is the central question of this study? Does the AGEs-RAGE axis mediate myocardial fibrosis in heart failure? What is the main finding and its importance? The AGEs-RAGE axis is involved in the pathogenesis of myocardial fibrosis through CFs activation induced by autophagy in heart failure. Inhibiting AGEs-RAGE axis attenuates dysfunction in the heart and attenuates myocardial fibrosis in mice with TAC via suppressing CFs activation. Abstract: Background: Heart failure is the end stage of cardiovascular diseases, and is a critical medical condition that poses an important therapeutic challenge for physicians due to its high morbidity and mortality. Myocardial fibrosis is part of the remodeling process that occurs in heart failure. Many studies have shown that advanced glycation end products (AGEs) and receptor for advanced glycation end products (RAGE) are implicated in fibrosis and autophagy, but the mechanism remains unclear. In this study, we elucidate the mechanism by which the AGEs-RAGE axis mediates activation of cardiac fibroblasts (CFs) in heart failure. Methods and results: We used C57BL/6J wild-type (WT) mice to establish a model of heart failure by transverse aortic constriction (TAC). After 6 weeks of treatment, relevant indicators were detected. In mice subjected to TAC, AGEs were upregulated compared with sham mice. Inhibition of RAGE resulted in functional cardiac protection with reduced hypertrophy and fibrosis in mice after TAC. Of note, autophagy mediated the activation of CFs that transformed to myofibroblasts and contributed to fibrosis. In vitro, CFs were obtained from neonatal Sprague-Dawley rats and treated with AGEs-BSA and short hairpin RNA (shRNA) for RAGE, which to verify the results in vivo. Conclusions: These results suggest that the AGEs-RAGE axis is involved in the pathogenesis of myocardial fibrosis through CF activation induced by autophagy in heart failure. Inhibiting the AGEs-RAGE axis attenuates dysfunction in the heart and attenuates myocardial fibrosis in mice with TAC via suppressing CF activation. This article is protected by copyright. All rights reserved.
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Intestinal fibrosis is an important complication of inflammatory bowel disease (IBD). In the course of the development of fibrosis, certain parts of the intestine become narrowed, significantly destroying the structure and function of the intestine and affecting the quality of life of patients. Chronic inflammation is an important initiating factor of fibrosis. Unfortunately, the existing anti-inflammatory drugs cannot effectively prevent and alleviate fibrosis, and there is no effective anti-fibrotic drug, which makes surgical treatment the mainstream treatment for intestinal fibrosis and stenosis. Mesenchymal stem cells (MSCs) are capable of tissue regeneration and repair through their self-differentiation, secretion of cytokines, and secretion of extracellular vesicles. MSCs have been shown to play an important therapeutic role in the fibrosis of many organs. However, the role of MSC in intestinal fibrosis largely remained unexplored. This review summarizes the mechanism of intestinal fibrosis, including the role of immune cells, TGF-β, and the gut microbiome and metabolites. Available treatment options for fibrosis, particularly, MSCs are also discussed.
Chapter
The major distinguishing feature of fibrosis is significant deposition of collagen and other extracellular matrix (ECM) proteins, which can result in scarring if sufficiently excessive. Fibrosis affects many tissue types, and thus contributes to a broad group of diseases which, with few exceptions, continues to lack specific therapy. It has been estimated that nearly 45% of deaths in the developed world are caused by fibroproliferative diseases, which contribute to cardiovascular disease, pulmonary, renal, gut and liver fibrosis, and scleroderma (Bitterman and Henke in Chest 99:81S–84S, [1]). Fibroblasts are the most common stromal cell type of the connective tissues found in the body, and are the primary source of ECM in physiological conditions, i.e. in the absence of disease. The conversion of fibroblasts or similar stromal cells to myofibroblasts is a principal mediator of pathological fibrosis in many tissue types, and frequently occurs in response to ongoing tissue injury and chronic inflammation. While the fibrotic response can occur in response to existing disease, the phenotype conversion of fibroblasts to myofibroblasts due to transient stress or damage may lead to the initiation of long-term fibrotic disease (Bagchi et al. in BMC Biol 14:21, [2]). Inflammation has been found to be a critical inducer of fibrosis, with immune cells generating a variety of growth factors and cytokines that play critical roles in fibroblast activation and subsequent tissue remodelling and fibrosis. A common cellular response to stress stimuli such as inflammation is autophagy, and recent studies have tightly linked the activation or inhibition of autophagy with fibrotic diseases in myriad tissues. Here, we discuss the inter-relationship of these pathways to provide insight into their potential as therapeutic targets in fibrotic disease.