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Article ; Online: Lipid Peroxidation as the Mechanism Underlying Polycyclic Aromatic Hydrocarbons and Sunlight Synergistic Toxicity in Dermal Fibroblasts.

Larnac, Eloïse / Montoni, Alicia / Haydont, Valérie / Marrot, Laurent / Rochette, Patrick J

International journal of molecular sciences

2024 Volume 25, Issue 3

Abstract: Light and atmospheric pollution are both independently implicated in cancer induction and premature aging. Evidence has been growing more recently on the toxic synergy between light and pollutants. Polycyclic aromatic hydrocarbons (PAHs) originate from ... ...

Abstract	Light and atmospheric pollution are both independently implicated in cancer induction and premature aging. Evidence has been growing more recently on the toxic synergy between light and pollutants. Polycyclic aromatic hydrocarbons (PAHs) originate from the incomplete combustion of organic matter. Some PAHs, such as the Benzo[a]pyrene (BaP), absorb ultraviolet A (UVA) wavelengths and can act as exogenous chromophores, leading to synergistic toxicity through DNA damage and cytotoxicity concomitant to ROS formation. In this study, we shed light on the mechanism underlying the toxic synergy between PAHs and UVA. Using dermal fibroblasts co-exposed to UVA and BaP, we have demonstrated that the photosensitization reaction causes mortality, which is most likely caused by ROS accumulation. We have shown that these ROS are concentrated in the lipids, which causes an important induction of lipid peroxidation and malondialdehyde, by-products of lipid peroxidation. We have also shown the accumulation of bulky DNA damage, most likely generated by these by-products of lipid peroxidation. To our knowledge, this study represents the first one depicting the molecular effects of photo-pollution on dermal skin.
MeSH term(s)	Lipid Peroxidation ; Polycyclic Aromatic Hydrocarbons/toxicity ; Reactive Oxygen Species ; Ultraviolet Rays ; Sunlight/adverse effects ; Benzo(a)pyrene ; Fibroblasts
Chemical Substances	Polycyclic Aromatic Hydrocarbons ; Reactive Oxygen Species ; Benzo(a)pyrene (3417WMA06D)
Language	English
Publishing date	2024-02-05
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms25031905
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Article ; Online: Age-related evolutions of the dermis: Clinical signs, fibroblast and extracellular matrix dynamics.

Haydont, Valérie / Bernard, Bruno A / Fortunel, Nicolas O

Mechanisms of ageing and development

2018 Volume 177, Page(s) 150–156

Abstract: Ageing is today a major societal concern that is intrinsically associated with the increase of life expectancy. Outside the context of severe degenerative diseases that affect the elderly populations, normal visible signs of ageing, notably skin sagging ... ...

Abstract	Ageing is today a major societal concern that is intrinsically associated with the increase of life expectancy. Outside the context of severe degenerative diseases that affect the elderly populations, normal visible signs of ageing, notably skin sagging and wrinkles, influence the social and individual perception of peoples. Accordingly, there is a strong demand for researches on skin ageing. Deciphering the cellular and molecular processes of skin evolution through ageing is thus an active scientific domain, at the frontier of tissue developmental and ageing biology. The focus of the present article is to provide an overview of the current knowledge concerning the evolution of dermis characteristics at different life stages, from intra-uterine to post-natal life. The description will integrate stage-specific and age-related changes in dermis characteristics at the tissue, cell, and molecular levels.
MeSH term(s)	Aging/metabolism ; Aging/pathology ; Animals ; Dermis/metabolism ; Dermis/pathology ; Extracellular Matrix/metabolism ; Extracellular Matrix/pathology ; Humans ; Skin Aging
Language	English
Publishing date	2018-03-13
Publishing country	Ireland
Document type	Journal Article ; Review
ZDB-ID	183915-9
ISSN	1872-6216 ; 0047-6374
ISSN (online)	1872-6216
ISSN	0047-6374
DOI	10.1016/j.mad.2018.03.006
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Article ; Online: Transcriptome profiling of human papillary and reticular fibroblasts from adult interfollicular dermis pinpoints the 'tissue skeleton' gene network as a component of skin chrono-ageing.

Haydont, Valérie / Neiveyans, Véronique / Fortunel, Nicolas O / Asselineau, Daniel

Mechanisms of ageing and development

2019 Volume 179, Page(s) 60–77

Abstract: Interactions between extracellular matrix (ECM) and fibroblasts are essential for maintaining dermis integrity, and are subject to ageing. The ötissue skeleton' network connects ECM to the nucleus and DNA, impacting nuclear shape and gene expression. In ... ...

Abstract	Interactions between extracellular matrix (ECM) and fibroblasts are essential for maintaining dermis integrity, and are subject to ageing. The ötissue skeleton' network connects ECM to the nucleus and DNA, impacting nuclear shape and gene expression. In a previous Mech Ageing Dev publication, we have presented a transcriptomic study of papillary (Fp) and reticular (Fr) fibroblasts, with a main focus on Fp ageing. As shown here, ageing affects ötissue skeleton' transcripts, even more clearly in Fr than in Fp. Accordingly, using circular index measurement, we show that nuclear shape is affected by ageing in both cell fractions.
MeSH term(s)	Adult ; Aging ; Dermis/pathology ; Extracellular Matrix/metabolism ; Fibroblasts/metabolism ; Gene Expression Profiling/methods ; Gene Expression Regulation ; Gene Regulatory Networks ; Humans ; Skin/pathology ; Skin Aging/pathology ; Transcription, Genetic ; Transcriptome
Language	English
Publishing date	2019-01-11
Publishing country	Ireland
Document type	Journal Article
ZDB-ID	183915-9
ISSN	1872-6216 ; 0047-6374
ISSN (online)	1872-6216
ISSN	0047-6374
DOI	10.1016/j.mad.2019.01.003
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Article ; Online: NRF2 Shortage in Human Skin Fibroblasts Dysregulates Matrisome Gene Expression and Affects Collagen Fibrillogenesis.

Salamito, Mélanie / Gillet, Benjamin / Syx, Delfien / Vaganay, Elisabeth / Malbouyres, Marilyne / Cerutti, Catherine / Tissot, Nicolas / Exbrayat-Héritier, Chloé / Perez, Philippe / Jones, Christophe / Hughes, Sandrine / Malfait, Fransiska / Haydont, Valérie / Jäger, Sibylle / Ruggiero, Florence

The Journal of investigative dermatology

2022 Volume 143, Issue 3, Page(s) 386–397.e12

Abstract: NRF2 is a master regulator of the antioxidative response that was recently proposed as a potential regulator of extracellular matrix (ECM) gene expression. Fibroblasts are major ECM producers in all connective tissues, including the dermis. A better ... ...

Abstract	NRF2 is a master regulator of the antioxidative response that was recently proposed as a potential regulator of extracellular matrix (ECM) gene expression. Fibroblasts are major ECM producers in all connective tissues, including the dermis. A better understanding of NRF2-mediated ECM regulation in skin fibroblasts is thus of great interest for skin homeostasis maintenance and aging protection. In this study, we investigate the impact of NRF2 downregulation on matrisome gene expression and ECM deposits in human primary dermal fibroblasts. RNA-sequencing‒based transcriptome analysis of NRF2 silenced dermal fibroblasts shows that ECM genes are the most regulated gene sets, highlighting the relevance of the NRF2-mediated matrisome program in these cells. Using complementary light and electron microscopy methods, we show that NRF2 deprivation in dermal fibroblasts results in reduced collagen I biosynthesis and impacts collagen fibril deposition. Moreover, we identify ZNF469, a putative transcriptional regulator of collagen biosynthesis, as a target of NRF2. Both ZNF469 silenced fibroblasts and fibroblasts derived from Brittle Corneal Syndrome patients carrying variants in ZNF469 gene show reduced collagen I gene expression. Our study shows that NRF2 orchestrates matrisome expression in human skin fibroblasts through direct or indirect transcriptional mechanisms that could be prioritized to target dermal ECM homeostasis in health and disease.
MeSH term(s)	Humans ; NF-E2-Related Factor 2/genetics ; NF-E2-Related Factor 2/metabolism ; Extracellular Matrix/metabolism ; Collagen/metabolism ; Collagen Type I/genetics ; Collagen Type I/metabolism ; Gene Expression ; Fibroblasts/metabolism ; Cells, Cultured
Chemical Substances	NF-E2-Related Factor 2 ; Collagen (9007-34-5) ; Collagen Type I
Language	English
Publishing date	2022-09-20
Publishing country	United States
Document type	Journal Article
ZDB-ID	80136-7
ISSN	1523-1747 ; 0022-202X
ISSN (online)	1523-1747
ISSN	0022-202X
DOI	10.1016/j.jid.2022.07.034
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Article ; Online: Fibroblasts from the Human Skin Dermo-Hypodermal Junction are Distinct from Dermal Papillary and Reticular Fibroblasts and from Mesenchymal Stem Cells and Exhibit a Specific Molecular Profile Related to Extracellular Matrix Organization and Modeling.

Haydont, Valérie / Neiveyans, Véronique / Perez, Philippe / Busson, Élodie / Lataillade, JeanJacques / Asselineau, Daniel / Fortunel, Nicolas O

Cells

2020 Volume 9, Issue 2

Abstract: Human skin dermis contains fibroblast subpopulations in which characterization is crucial due to their roles in extracellular matrix (ECM) biology. This study investigates the properties of fibroblasts localized at the frontier of deep dermis and ... ...

Abstract	Human skin dermis contains fibroblast subpopulations in which characterization is crucial due to their roles in extracellular matrix (ECM) biology. This study investigates the properties of fibroblasts localized at the frontier of deep dermis and hypodermis, i.e., dermo-hypodermal junction fibroblasts (F-DHJ), which were compared to intermediate reticular dermis (Fr) and superficial papillary dermis (Fp) fibroblasts. F-DHJ differed from Fr and Fp cells in their wider potential for differentiation into mesodermal lineages and in their absence of contractility when integrated in a three-dimensional dermal equivalent. The transcriptomic profile of F-DHJ exhibited specificities in the expression of genes involved in ECM synthesis-processing and "tissue skeleton" organization. In accordance with transcriptome data, ECM proteins, notably Tenascin C, distributions differed between the reticular dermis and the dermo-hypodermal junction areas, which was documented in normal adult skin. Finally, genome-wide transcriptome profiling was used to evaluate the molecular proximity of F-DHJ with the two dermal fibroblast populations (Fp and Fr) and with the mesenchymal stem cells (MSCs) corresponding to five tissue origins (bone marrow, fat, amnion, chorion, and cord). This comparative analysis classified the three skin fibroblast types, including F-DHJ, as a clearly distinct group from the five MSC sample origins.
MeSH term(s)	Adolescent ; Adult ; Aged ; Aging/physiology ; Biomarkers/metabolism ; Cell Differentiation ; Cell Lineage ; Cell Proliferation ; Cell Shape ; Dermis/cytology ; Extracellular Matrix/metabolism ; Female ; Fibroblasts/cytology ; Fibroblasts/metabolism ; Humans ; Mesenchymal Stem Cells/cytology ; Mesenchymal Stem Cells/metabolism ; Middle Aged ; Models, Biological ; Phenotype ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Tenascin/metabolism ; Transcriptome/genetics ; Young Adult
Chemical Substances	Biomarkers ; RNA, Messenger ; Tenascin
Language	English
Publishing date	2020-02-05
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2661518-6
ISSN	2073-4409 ; 2073-4409
ISSN (online)	2073-4409
ISSN	2073-4409
DOI	10.3390/cells9020368
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Article ; Online: Genome-wide profiling of adult human papillary and reticular fibroblasts identifies ACAN, Col XI α1, and PSG1 as general biomarkers of dermis ageing, and KANK4 as an exemplary effector of papillary fibroblast ageing, related to contractility.

Haydont, Valérie / Neiveyans, Véronique / Zucchi, Hélène / Fortunel, Nicolas O / Asselineau, Daniel

Mechanisms of ageing and development

2018 Volume 177, Page(s) 157–181

Abstract: Deciphering the characteristics of dermal fibroblasts is critical to further understand skin ageing. We have conducted a genome-wide transcriptomic characterization of papillary (Fp) and reticular (Fr) fibroblasts extracted from human skin samples ... ...

Abstract	Deciphering the characteristics of dermal fibroblasts is critical to further understand skin ageing. We have conducted a genome-wide transcriptomic characterization of papillary (Fp) and reticular (Fr) fibroblasts extracted from human skin samples corresponding to younger and older adult ages. From this screen, biomarkers suitable for the assessment of chronological ageing were identified, and extrapolated to the context of photo-damaged skin. In particular, KANK4, ACAN, Col XI α1, and PSG1, were expressed at an increased level in both chronologically-aged and photo-damaged skin. Notably, analysis focused on Fp identified significant transcriptional signatures associated with ageing, which included transcripts related to extracellular matrix, focal adhesion points, and cytoskeleton, thus suggesting functional consequences on tissue structure. At a cellular level, an increased contractility was identified as a property of aged Fp. Accordingly, further investigations were conducted on the KN motif and ankyrin repeat-containing protein 4 (KANK4) to explore its possible function as an original effector involved in the acquisition of aged properties in Fp, notably their increased contractility. We show that KANK4 down-modulation using siRNA led to increased Rho pathway activity, thereby reducing their contractility. As a proof-of-principle, the present study shows that targeting KANK4 was efficient to attenuate aged Fp characteristics.
MeSH term(s)	Adolescent ; Adult ; Aged ; Aggrecans/metabolism ; Carrier Proteins/metabolism ; Cellular Senescence ; Collagen Type X/metabolism ; Female ; Fibroblasts/metabolism ; Fibroblasts/pathology ; Genome-Wide Association Study ; Humans ; Middle Aged ; Skin Aging
Chemical Substances	ACAN protein, human ; Aggrecans ; Carrier Proteins ; Collagen Type X
Language	English
Publishing date	2018-06-18
Publishing country	Ireland
Document type	Journal Article
ZDB-ID	183915-9
ISSN	1872-6216 ; 0047-6374
ISSN (online)	1872-6216
ISSN	0047-6374
DOI	10.1016/j.mad.2018.06.003
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Article ; Online: Rho/ROCK pathway as a molecular target for modulation of intestinal radiation-induced toxicity.

Haydont, V / Bourgier, C / Vozenin-Brotons, M-C

The British journal of radiology

2007 Volume 80 Spec No 1, Page(s) S32–40

Abstract: More than half of cancer patients are treated with radiation therapy. Despite its high therapeutic index, radiation therapy can cause disabling injuries to normal tissues, especially in long-term survivors. Thus, one of the great challenges of modern ... ...

Abstract	More than half of cancer patients are treated with radiation therapy. Despite its high therapeutic index, radiation therapy can cause disabling injuries to normal tissues, especially in long-term survivors. Thus, one of the great challenges of modern radiation therapy is to increase tolerance of normal tissue to ionizing radiation in order to improve the quality of life of cancer survivors and/or enhance local control using dose escalation. The physiopathological aspects of normal tissue toxicity have been widely explored; however, none of these descriptive findings has led to the development of effective therapeutic strategies. Several empirical treatments have also been used in clinical trials (superoxide dismutase, pentoxifylline-tocopherol); however, the results are still controversial, and their mechanisms of action have not been clearly defined. The recent development of high-throughput biological approaches will contribute greatly to the characterization of the molecular pathways associated with normal tissue toxicity and the identification of specific and effective molecular targets for therapeutic interventions using already known or new pharmacological compounds. In this paper, we will discuss recent advances made in the characterization of one of the most serious complications of radiation therapy, late intestinal toxicity, using molecular profiling. We will focus on the involvement of the Rho/ROCK pathway in the development and maintenance of late radiation enteropathy. The role of the Rho/ROCK pathway in tissue response to radiation injury will be reviewed, as well as therapeutic perspectives.
MeSH term(s)	Animals ; Connective Tissue Growth Factor ; Fibrosis/etiology ; Fibrosis/physiopathology ; Humans ; Immediate-Early Proteins/physiology ; Intercellular Signaling Peptides and Proteins/physiology ; Intestines/pathology ; Intestines/radiation effects ; Intracellular Signaling Peptides and Proteins/antagonists & inhibitors ; Intracellular Signaling Peptides and Proteins/physiology ; Protein-Serine-Threonine Kinases/antagonists & inhibitors ; Protein-Serine-Threonine Kinases/physiology ; Radiation Injuries/drug therapy ; Radiation Injuries/physiopathology ; Signal Transduction/drug effects ; Signal Transduction/physiology ; rho-Associated Kinases
Chemical Substances	CCN2 protein, human ; Immediate-Early Proteins ; Intercellular Signaling Peptides and Proteins ; Intracellular Signaling Peptides and Proteins ; Connective Tissue Growth Factor (139568-91-5) ; Protein-Serine-Threonine Kinases (EC 2.7.11.1) ; rho-Associated Kinases (EC 2.7.11.1)
Language	English
Publishing date	2007-06-18
Publishing country	England
Document type	Journal Article ; Review
ZDB-ID	2982-8
ISSN	1748-880X ; 0007-1285
ISSN (online)	1748-880X
ISSN	0007-1285
DOI	10.1259/bjr/58514380
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Article: Maintenance of radiation-induced intestinal fibrosis: cellular and molecular features.

Haydont, Valérie / Vozenin-Brotons, Marie-Catherine

World journal of gastroenterology

2007 Volume 13, Issue 19, Page(s) 2675–2683

Abstract: Recent advances in cell and molecular radiobiology clearly showed that tissue response to radiation injury cannot be restricted to a simple cell-killing process, but depends upon continuous and integrated pathogenic processes, involving cell ... ...

Abstract	Recent advances in cell and molecular radiobiology clearly showed that tissue response to radiation injury cannot be restricted to a simple cell-killing process, but depends upon continuous and integrated pathogenic processes, involving cell differentiation and crosstalk between the various cellular components of the tissue within the extracellular matrix. Thus, the prior concept of primary cell target in which a single-cell type (whatever it's epithelial or endothelial cells) dictates the whole tissue response to radiation injury has to be replaced by the occurrence of coordinated multicellular response that may either lead to tissue recovery or to sequel development. In this context, the present review will focus on the maintenance of the radiation-induced wound healing and fibrogenic signals triggered by and through the microenvironment toward the mesenchymal cell compartment, and will highlight how sequential and sustained modifications in cell phenotypes will in cascade modify cell-to-cell interactions and tissue composition.
MeSH term(s)	Cell Communication ; Cell Differentiation/radiation effects ; Fibrosis ; Humans ; Intestines/pathology ; Intestines/radiation effects ; Mesoderm/pathology ; Mesoderm/radiation effects ; Radiation Injuries/pathology ; Radiotherapy/adverse effects
Language	English
Publishing date	2007-06-06
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2185929-2
ISSN	2219-2840 ; 1007-9327
ISSN (online)	2219-2840
ISSN	1007-9327
DOI	10.3748/wjg.v13.i19.2675
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Zs.A 6039: Show issues

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Article: Specific signals involved in the long-term maintenance of radiation-induced fibrogenic differentiation: a role for CCN2 and low concentration of TGF-beta1.

Haydont, Valérie / Riser, Bruce L / Aigueperse, Jocelyne / Vozenin-Brotons, Marie-Catherine

American journal of physiology. Cell physiology

2008 Volume 294, Issue 6, Page(s) C1332–41

Abstract: The fibrogenic differentiation of resident mesenchymal cells is a key parameter in the pathogenesis of radiation fibrosis and is triggered by the profibrotic growth factors transforming growth factor (TGF)-beta1 and CCN2. TGF-beta1 is considered the ... ...

Abstract	The fibrogenic differentiation of resident mesenchymal cells is a key parameter in the pathogenesis of radiation fibrosis and is triggered by the profibrotic growth factors transforming growth factor (TGF)-beta1 and CCN2. TGF-beta1 is considered the primary inducer of fibrogenic differentiation and is thought to control its long-term maintenance, whereas CCN2 is considered secondary effector of TGF-beta1. Yet, in long-term established fibrosis like that associated with delayed radiation enteropathy, in situ TGF-beta1 deposition is low, whereas CCN2 expression is high. To explore this apparent paradox, cell response to increasing doses of TGF-beta1 was investigated in cells modeling initiation and maintenance of fibrosis, i.e., normal and fibrosis-derived smooth muscle cells, respectively. Activation of cell-specific signaling pathways by low TGF-beta1 doses was demonstrated with a main activation of the Rho/ROCK pathway in fibrosis-derived cells, whereas the Smad pathway was mainly activated in normal cells. This leads to subsequent and cell-specific regulation of the CCN2 gene. These results suggested a specific profibrotic role of CCN2 in fibrosis-initiated cells. Furthermore, the modulation of CCN2 expression by itself and the combination of TGF-beta1 and CCN2 was investigated in fibrosis-derived cells. In fibrosis-initiated cells CCN2 triggered its autoinduction; furthermore, low concentration of TGF-beta1-potentiated CCN2 autoinduction. Our findings showed a differential requirement and action of TGF-beta1 in the fibrogenic response of normal vs. fibrosis-derived cells. This study defines a novel Rho/ROCK but Smad3-independent mode of TGF-beta signaling that may operate during the chronic stages of fibrosis and provides evidence of both specific and combinatorial roles of low TGF-beta1 dose and CCN2.
MeSH term(s)	Aged ; Cell Differentiation/radiation effects ; Cells, Cultured ; Colon/enzymology ; Colon/metabolism ; Colon/pathology ; Colon/radiation effects ; Connective Tissue Growth Factor ; Enzyme Activation ; Fibrosis ; Humans ; Ileum/metabolism ; Ileum/pathology ; Immediate-Early Proteins/genetics ; Immediate-Early Proteins/metabolism ; Intercellular Signaling Peptides and Proteins/genetics ; Intercellular Signaling Peptides and Proteins/metabolism ; Intestinal Diseases/etiology ; Intestinal Diseases/metabolism ; Intestinal Diseases/pathology ; Middle Aged ; Myocytes, Smooth Muscle/metabolism ; Myocytes, Smooth Muscle/pathology ; Myocytes, Smooth Muscle/radiation effects ; Radiation Injuries/etiology ; Radiation Injuries/metabolism ; Radiation Injuries/pathology ; Radiotherapy/adverse effects ; Recombinant Proteins/metabolism ; Signal Transduction/radiation effects ; Smad Proteins/metabolism ; Transcriptional Activation ; Transforming Growth Factor beta1/metabolism ; rho-Associated Kinases/metabolism
Chemical Substances	CCN2 protein, human ; Immediate-Early Proteins ; Intercellular Signaling Peptides and Proteins ; Recombinant Proteins ; Smad Proteins ; Transforming Growth Factor beta1 ; Connective Tissue Growth Factor (139568-91-5) ; rho-Associated Kinases (EC 2.7.11.1)
Language	English
Publishing date	2008-04-09
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	392098-7
ISSN	1522-1563 ; 0363-6143
ISSN (online)	1522-1563
ISSN	0363-6143
DOI	10.1152/ajpcell.90626.2007
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Article: Inhibition of Rho kinase modulates radiation induced fibrogenic phenotype in intestinal smooth muscle cells through alteration of the cytoskeleton and connective tissue growth factor expression.

Bourgier, C / Haydont, V / Milliat, F / François, A / Holler, V / Lasser, P / Bourhis, J / Mathé, D / Vozenin-Brotons, M-C

Gut

2005 Volume 54, Issue 3, Page(s) 336–343

Abstract: Background: Late radiation enteritis in humans is associated with accumulation of extracellular matrix and increased connective tissue growth factor (CTGF) expression that may involve intestinal muscular layers.: Aims: We investigated the molecular ... ...

Abstract	Background: Late radiation enteritis in humans is associated with accumulation of extracellular matrix and increased connective tissue growth factor (CTGF) expression that may involve intestinal muscular layers. Aims: We investigated the molecular pathways involved in maintenance of radiation induced fibrosis by gene profiling and postulated that alteration of the Rho pathway could be associated with radiation induced fibrogenic signals and CTGF sustained expression. Patients and methods: Ileal biopsies from individuals with and without radiation enteritis were analysed by cDNA array, and primary cultures of intestinal smooth muscle cells were established. Then, the effect of pharmacological inhibition of p160 Rho kinase, using Y-27632, was studied by real time reverse transcription-polymerase chain reaction, western blot, and electrophoretic mobility shift assay. Results: Molecular profile analysis of late radiation enteritis showed alterations in expression of genes coding for the Rho proteins. To investigate further the involvement of the Rho pathway in intestinal radiation induced fibrosis, primary intestinal smooth muscle cells were isolated from radiation enteritis. They retained their fibrogenic differentiation in vitro, exhibited a typical cytoskeletal network, a high constitutive CTGF level, increased collagen secretory capacity, and altered expression of genes coding for the Rho family. Rho kinase blockade induced a simultaneous decrease in the number of actin stress fibres, alpha smooth muscle actin, and heat shock protein 27 levels. It also decreased CTGF levels, probably through nuclear factor kappaB inhibition, and caused decreased expression of the type I collagen gene. Conclusion: This study is the first showing involvement of the Rho/Rho kinase pathway in radiation fibrosis and intestinal smooth muscle cell fibrogenic differentiation. It suggests that specific inhibition of Rho kinase may be a promising approach for the development of antifibrotic therapies.
MeSH term(s)	Actins/metabolism ; Adult ; Aged ; Aged, 80 and over ; Amides/pharmacology ; Cell Differentiation ; Cells, Cultured ; Connective Tissue Growth Factor ; Cytoskeleton/metabolism ; Cytoskeleton/radiation effects ; DNA-Binding Proteins/metabolism ; Enteritis/enzymology ; Enteritis/etiology ; Enteritis/pathology ; Enzyme Inhibitors/pharmacology ; Female ; Fibrosis/etiology ; Fibrosis/pathology ; Gene Expression Profiling/methods ; Humans ; Ileum/pathology ; Immediate-Early Proteins/metabolism ; Intercellular Signaling Peptides and Proteins/metabolism ; Intracellular Signaling Peptides and Proteins ; Male ; Middle Aged ; Muscle, Smooth/metabolism ; Muscle, Smooth/pathology ; Muscle, Smooth/radiation effects ; NF-kappa B/pharmacology ; Protein-Serine-Threonine Kinases/antagonists & inhibitors ; Protein-Serine-Threonine Kinases/physiology ; Pyridines/pharmacology ; Radiation Injuries/enzymology ; Radiation Injuries/etiology ; Radiation Injuries/pathology ; Reverse Transcriptase Polymerase Chain Reaction/methods ; Signal Transduction ; rho GTP-Binding Proteins/physiology ; rho-Associated Kinases
Chemical Substances	Actins ; Amides ; CCN2 protein, human ; DNA-Binding Proteins ; Enzyme Inhibitors ; Immediate-Early Proteins ; Intercellular Signaling Peptides and Proteins ; Intracellular Signaling Peptides and Proteins ; NF-kappa B ; Pyridines ; Y 27632 (138381-45-0) ; Connective Tissue Growth Factor (139568-91-5) ; Protein-Serine-Threonine Kinases (EC 2.7.11.1) ; rho-Associated Kinases (EC 2.7.11.1) ; rho GTP-Binding Proteins (EC 3.6.5.2)
Language	English
Publishing date	2005-02-09
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	80128-8
ISSN	1468-3288 ; 0017-5749
ISSN (online)	1468-3288
ISSN	0017-5749
DOI	10.1136/gut.2004.051169
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