Hsp47 Gene Research Articles

Growth performance, hematological profile, and related genes expression in goldfish (Carassius auratus) fed on rosmarinic acid-enriched diets and subjected to ambient ammonia

Recently, supplementing fish diets with polyphenols has attracted more attention because of their great nutritional role. The unionized ammonia is one of the critical water parameters that adversely affects the fish performance and welfare status. Hence, the present perspective outlines the potential value of dietary rosmarinic acid (RA) in boosting growth, hematological, and immune biomarkers, besides related-genes expressions post-exposure of goldfish (Carassius auratus) to ambient ammonia. Fish (n = 240) were separated into four groups; RA0 (the control), RA400, RA600, and RA800 received 0, 400, 600, and 800 mg RA/kg feed, respectively. After 56 days, counts of white and red blood cells and growth indices (weight gain, specific growth rate, feed intake, and feed conversion ratio) were evaluated. After that, the fish per each treatment were exposed to 0.5 mg/L of unionized ammonia for three hours. After the challenging time, many biomarkers were measured including immune response (ACH50, total immunoglobulin, and lysozymes activity), digestion (proteases activity), and genes expression involving MUC, HSP27, HSP30, HSP47, TNFa, Lp1, and Fas genes plus the anti-inflammatory cytokines (IL6, IL8, and IL10). The findings of the current research revealed that dietary RA intervention fostered growth performance, improved blood picture (p < 0.05), and enhanced (p < 0.05) the expression of genes, which are responsible for modulating immunity and anti-inflammatory status in a dose-dependent manner. The present study concluded that dietary RA is a successful natural feed supplement for goldfish with an optimal level of 600 mg RA/kg feed. However, it verifies antitoxic and anti-inflammatory properties against ambient ammonia toxicity. It also has a promising influence on the growth, hematological, and immunity and anti-inflammatory related genes expression of goldfish, C. auratus.

HSP47 levels determine the degree of body adiposity

Adiposity varies among individuals with the influence of diverse physiological, pathological, environmental, hormonal, and genetic factors, but a unified molecular basis remains elusive. Here, we identify HSP47, a collagen-specific chaperone, as a key determinant of body adiposity. HSP47 expression is abundant in adipose tissue; increased with feeding, overeating, and obesity; decreased with fasting, exercise, calorie restriction, bariatric surgery, and cachexia; and correlated with fat mass, BMI, waist, and hip circumferences. Insulin and glucocorticoids, respectively, up- and down-regulate HSP47 expression. In humans, the increase of HSP47 gene expression by its intron or synonymous variants is associated with higher body adiposity traits. In mice, the adipose-specific knockout or pharmacological inhibition of HSP47 leads to lower body adiposity compared to the control. Mechanistically, HSP47 promotes collagen dynamics in the folding, secretion, and interaction with integrin, which activates FAK signaling and preserves PPARγ protein from proteasomal degradation, partly related to MDM2. The study highlights the significance of HSP47 in determining the amount of body fat individually and under various circumstances.

Single Nucleotide Polymorphisms of the MEFV Gene E148Q Are Highly Associated With Disease Phenotype in Crohn's Disease.

Single nucleotide polymorphisms (SNPs) of the MEFV gene may modify inflammatory bowel disease (IBD) activity. The prevalence of MEFV gene SNPs in IBD patients and their involvement in IBD pathophysiology remains unclear. We analyzed 12 MEFV gene SNPs in peripheral leukocytes of Japanese IBD patients (Crohn's disease [CD]: 69 patients, ulcerative colitis: 32 patients) by polymerase chain reaction using next-generation DNA sequencing and evaluated their prevalence and association with the disease characteristics. Inflammasome activity and mature interleukin (IL)-1β and IL-18 production were evaluated in peripheral blood mononuclear cells obtained from CD patients stimulated with lipopolysaccharides and adenosine triphosphate, and compared between those with and without the E148Q SNP. COL1A1 and HSP47 gene expression was analyzed in CCD-18Co cells costimulated with IL-1β and other inflammatory cytokines. The prevalence of MEFV gene SNPs in IBD patients was similar to that in the human gene database. E148Q was the most common SNP. Compared with CD patients without E148Q, those with E148Q had a significantly greater frequency of the stricture phenotype, and their peripheral blood mononuclear cells exhibited significantly higher IL-1β and IL-18 levels and higher caspase-1 activity. IL-1β and IL-17A synergistically increased COL1A1 and HSP47 gene expression. MEFV gene SNPs, including E148Q, modify the behavior of CD. IL-1β and IL-18 are produced through enhanced caspase-1 activity in monocytes of CD patients with E148Q. IL-1β promotes gene expression of fibrosis-related genes by cooperating with IL-17A in myofibroblasts. Therefore, E148Q might be a disease-modifying gene associated with the fibrostenosis phenotype in CD patients.

Molecular Characterization of Hsp47 in Grass Carp (Ctenopharyngodon idella) and Its Correlation with Type I Collagen in Response to Fish Aerobic Exercise

Heat shock protein 47 (Hsp47) is a collagen-specific molecular chaperone that is indispensable for molecular maturation of collagen. In this study, hsp47 and hsp47-like cDNAs were cloned and characterized in grass carp (Ctenopharyngodon idella). The cDNAs were 1212 and 1218 base pairs long, respectively, and included an open reading frame encoding 403 and 405 amino acids. The molecular phylogeny based on the deduced amino acid sequences indicated that the correct sequences of the hsp47 and hsp47-like cDNA were obtained and the deduced proteins clustered distinctly into teleost clades. Primary structure analysis and characterization of Hsp47 and Hsp47-like shared the basic structure and biofunctions of Hsp47 in vertebrates. The spatial pattern of gene expression revealed that hsp47 and hsp47-like were relatively ubiquitous in different tissues and highly expressed in heart and skin. The expression levels of hsp47 and hsp47-like and type I collagen mRNAs varied similarly in different tissues. Type I collagen content increased significantly with the increase of water velocity in the muscle of grass carp in response to aerobic exercise. Among the gene expression changes of hsp47, hsp47-like, col1a1 and col1a2 in muscle that occurred in response to aerobic exercise, the change of type I collagen was most strongly correlated with hsp47 expression. Additionally, col1a1 showed the highest correlation with hsp47-like and col1a2 showed the highest correlation with hsp47. These findings suggest that grass carp Hsp47 and Hsp47-like are closely related to type I collagen synthesis. This study firstly suggests fish aerobic exercise can improve type I collagen content and Hsp47 gene expression in muscle of grass carp.

Transcriptional and epigenetic effects of Vitis vinifera L. leaf extract on UV-stressed human dermal fibroblasts.

Adverse environmental conditions such as UV radiation induce oxidative and aging events leading to severe damage to human skin cells. Natural products such as plant extracts have been implicated in the skin anti-oxidant and anti-aging cellular protection against environmental stress. Moreover, environmental factors have been shown to impact chromatin structure leading to altered gene expression programs with profound changes in cellular functions. In this study, we assessed the in vitro effect of a leaf extract from Vitis vinifera L. on UV-stressed primary human dermal fibroblasts, focusing on gene expression and DNA methylation as an epigenetic factor. Expression analysis of two genes known to be implicated in skin anti-aging, SIRT1and HSP4, demonstrated significant induction in the presence of the extract under normal or UVA conditions. In addition, DNA methylation profiling of SIRT1 and HSP47 promoters showed that the V. vinifera L. extract induced changes in the DNA methylation pattern of both genes that may be associated with SIRT1 and HSP47 gene expression. Our study shows for the first time transcriptional and DNA methylation alterations on human skin fibroblasts exposed to UV stress and suggest a protective effect of a V. vinifera extract possibly through transcriptional regulation of critical skin anti-aging genes.

Temperature changes and marine fish species (Epinephelus coioides and Sparus aurata): Role of oxidative stress biomarkers in toxicological food studies

The increase of seawater temperature as a result of global climate variation elucidates a major challenge for marine organisms survival in addition to consumers safety. Spotted grouper (Epinephelus coioides) and Seabream (Sparus aurata) were collected in water with different temperature variations at Suez Canal and Alexandria (Suez and Abu Qir bay) in Egypt with the aim to assess expression levels of heat shock proteins such as HSP47, HSP70 and HSP90 genes in addition to antioxidants value through enzymes activity: Glutathione-S-Transferase (GST) and Glutathione Peroxidase (GPx). Research results revealed that expression of the HSP47, HSP70a and HSP90 genes increased in marine fishes tissues collected from Suez Canal, with higher water temperature (23:28ºC), compared with those collected from Alexandria (19:24°C) whereas the content of GPx and GST decreased. Our results show alteration of the marker examined suggesting that the increase of heat shock protein genes expression levels of fish collected from Suez Canal might be exposed mainly to thermal oxidative stress response more than those collect from Alexandria. The increase of heat shock protein-related genes expression could be considered as a factor in prohibiting the heat shock transcription factor that may lead to stimulation of heat-inducible genes in addition to heat acclimation. Thus, warming of water is also likely to alter the composition and abundance of food resources, e.g. fish muscles, available to higher trophic level consumers.

Abstract 2374: Mir-29/Hsp47 regulate breast cancer progression by modulating the ECM transcription network

Abstract The extracellular matrix (ECM), an essential component of tumor microenvironment, play crucial roles in cancer development and progression. Using gene co-expression network analysis, we have identified an ECM transcription network from hundreds of human breast cancer. In this report, we show how the molecular chaperone protein Hsp47 functions as a nodal hub in regulating an ECM gene transcription network. We found that Hsp47 expression was activated during breast cancer development and progression. Hsp47 silencing reprogrammed human breast cancer cells to form growth-arrested and/or non-invasive structures in 3D cultures, and to limit tumor growth in xenograft assays by reducing deposition of collagen and fibronectin. Co-expression network analysis also showed that levels of microRNA-29b and 29c were inversely correlated with expression of Hsp47 and ECM network genes in human breast cancer tissues. We found that miR-29 repressed expression of Hsp47 along with multiple ECM network genes. Ectopic expression of miR-29b suppressed malignant phenotypes of breast cancer cells in 3D culture. Clinically, increased expression of Hsp47 and reduced levels of miR-29b and 29c were associated with poor survival outcomes in breast cancer patients. Our results show that Hsp47 is regulated by miR-29 during breast cancer development and progression, and that increased Hsp47 expression promotes cancer progression in part by enhancing deposition of ECM proteins. Citation Format: Jieqing Zhu, Gaofeng Xiong, Ren Xu. Mir-29/Hsp47 regulate breast cancer progression by modulating the ECM transcription network. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2374. doi:10.1158/1538-7445.AM2015-2374

Overexpression of HSP47 in esophageal squamous cell carcinoma: clinical implications and functional analysis.

Several biomarkers of esophageal squamous cell carcinoma (ESCC) have been explored to improve the prognosis of this disease. One of these, the 47-kDa heat shock protein (HSP47), has been screened as a potential biomarker by genomic profiling and is known to be overexpressed in some malignant diseases. In this study, we explored the role and evaluated the prognostic value of HSP47 expression in ESCC. The function of this protein was analyzed by assaying proliferation, wound healing, and colony formation in an HSP47-knockdown ESCC line. The prognostic implication of HSP47 expression was analyzed by immunohistochemical staining in 157 surgical specimens. HSP47 expression level and other clinical variables were analyzed using multivariate Cox proportional hazards models. Silencing of the HSP47 gene in the ESCC cell line inhibited cell proliferation and colony formation. HSP47 was highly expressed in ESCC tissue samples, compared with normal esophageal tissues. The level of immunohistochemical staining of HSP47 and pathologic stage were significantly correlated with overall and recurrence-free survival, as shown by multivariate analysis (P = 0.014 and 0.044, respectively). We found that overexpression of HSP47 is associated with poor prognosis in patients with ESCC and that this is consistent with the function of HSP47 in terms of increased cell proliferation and colony formation. These results suggest that HSP47 is a potential prognostic biomarker for ESCC and merits further research for novel diagnostic and therapeutic applications.

Chaperone Hsp47 Drives Malignant Growth and Invasion by Modulating an ECM Gene Network

The extracellular matrix (ECM) is a determining factor in the tumor microenvironment that restrains or promotes malignant growth. In this report, we show how the molecular chaperone protein Hsp47 functions as a nodal hub in regulating an ECM gene transcription network. A transcription network analysis showed that Hsp47 expression was activated during breast cancer development and progression. Hsp47 silencing reprogrammed human breast cancer cells to form growth-arrested and/or noninvasive structures in 3D cultures, and to limit tumor growth in xenograft assays by reducing deposition of collagen and fibronectin. Coexpression network analysis also showed that levels of microRNA(miR)-29b and -29c were inversely correlated with expression of Hsp47 and ECM network genes in human breast cancer tissues. We found that miR-29 repressed expression of Hsp47 along with multiple ECM network genes. Ectopic expression of miR-29b suppressed malignant phenotypes of breast cancer cells in 3D culture. Clinically, increased expression of Hsp47 and reduced levels of miR-29b and -29c were associated with poor survival outcomes in breast cancer patients. Our results show that Hsp47 is regulated by miR-29 during breast cancer development and progression, and that increased Hsp47 expression promotes cancer progression in part by enhancing deposition of ECM proteins.

P078. Evaluating the efficacy of potential drugs for intestinal fibrosis using precision-cut tissue slices

Intestinal fibrosis (IF) is a common complication in Crohn's disease. Currently, there are no drugs registered to treat IF and the sole therapy is intestinal resection. Transforming growth factor (TGF)-beta and platelet-derived growth factor (PDGF) play a key role in IF and are the main targets for potential treatment. Recently, we developed a novel model for the early onset of IF using precision-cut intestinal slices (PCIS). Our objective was to investigate the antifibrotic effect of some potential antifibrotic compounds, including TGF-beta and PDGF-pathway inhibitors, by using the murine PCIS fibrosis model. Murine PCIS were incubated up to 48 h. The viability was assessed by evaluating the ATP content of the PCIS. Gene expression of the fibrosis markers pro-collagen 1a1 (Col1a1), heat shock protein 47 (Hsp47) and fibronectin (Fn2) were determined by qPCR. The effects of antifibrotic drugs mainly inhibiting the TGF-beta pathway: valproic acid (VPA), tetrandrine (Tet), pirfenidone (Pir), and LY2109761 (LY) and mainly inhibiting the PDGF pathway: imatinib (Ima), sorafenib (Sor), and sunitinib (Sun) were determined at the maximal non-toxic concentrations. Murine PCIS remained viable up to 48 h of incubation and showed increased gene expression of the fibrosis markers (Col1a1, 0.6; Hsp47, 4.0 and Fn2, 4.4 fold). After 48 h, VPA and Tet down-regulated Hsp47 gene expression 2.0 and 1.7 fold, respectively. Furthermore, Fn2 gene expression was also decreased 2.1 fold by Tet. Meanwhile, Pir decreased Col1a1, Hsp47, and Fn2 gene expression 2.2, 1.5, and 1.2 fold, respectively. All investigated markers of fibrosis were down-regulated by LY (Col1a1, 9.0; Hsp47, 1.9 and Fn2, 2.7 fold). Sun decreased the expression of Col1a1, 1.6; Hsp47, 3.3 and Fn2, 2.3 fold, while Sor only down-regulated Hsp47, 1.3 fold. In contrast, Ima did not affect the expression of fibrosis markers. From the compounds studied, the TGF-beta-inhibitors; Tet, Pir, and LY and only one PDGF-inhibitor, Sun, showed potential antifibrotic effect on gene expression of fibrosis markers in murine PCIS. Thus, PCIS is a promising model to evaluate the antifibrotic effect of potential drugs for intestinal fibrosis.

Deletion of the Collagen-specific Molecular Chaperone Hsp47 Causes Endoplasmic Reticulum Stress-mediated Apoptosis of Hepatic Stellate Cells

Chronic liver injury, often caused by alcoholism and viral hepatitis, causes liver fibrosis via the induction of collagen production. In liver fibrosis, hepatic stellate cells (HSCs) are activated and transform into myofibroblasts, which actively produce and secrete collagen into the extracellular matrix. Hsp47 (heat shock protein 47) is a collagen-specific molecular chaperone that is essential for the maturation and secretion of collagen. Here, we used the Cre-LoxP system to disrupt the Hsp47 gene in isolated HSCs from Hsp47 floxed mice. Immature type I procollagen accumulated and partially aggregated in Hsp47-KO HSCs. This accumulation was augmented when autophagy was inhibited, which induced expression of the endoplasmic reticulum (ER) stress-inducible proteins BiP (immunoglobulin heavy chain-binding protein) and Grp94 (94-kDa glucose-regulated protein). The inhibition of autophagy in Hsp47-KO HSCs also induced CHOP (CCAAT/enhancer-binding protein homologous protein), which is an ER stress-induced transcription factor responsible for apoptosis. These data suggest that apoptosis is induced through ER stress by procollagen accumulation in Hsp47-KO HSCs when autophagy is inhibited. Thus, Hsp47 could be a promising therapeutic target in liver fibrosis.

The Influence of Selected Flavonoids from the Leaves of Cirsium palustre (L.) Scop. on Collagen Expression in Human Skin Fibroblasts

Ten flavonoids belonging to the subclasses of flavones, flavanones and aurones were isolated from methanolic extract of Cirsium palustre leaves after multistep chromatographic separation. Their structures were elucidated with spectroscopic methods. All compounds, except for luteolin 7-O-glucoside, were isolated for the first time. Four compounds-eriodictyol 7-O-glucoside (6), 6-hydroxyluteolin 7-O-glucoside (11), scutellarein 7-O-glucoside (12) and pedalitin (14)-were tested for their effect on collagen expression in normal human dermal fibroblasts. Among them, compound 11 at 40 μM and compound 14, at all concentrations used (1, 20, 40 μM), significantly enhanced the level of total collagen secreted into the medium. Furthermore, compound 11 significantly stimulated type I collagen expression, whereas compound 14 activated type I and III collagen expression at the mRNA level, depending on concentration. MMP-2 activity was inhibited by all study compounds, with the greatest effect recorded with compound 14 at 20 μM. The lack of effect on collagen content in the medium of compound 6- and compound 12-treated cells, besides an increase in COL1A1 and COL1A2 expression, might be caused by diminished expression of HSP47 gene, resulting in decreased procollagen secretion. Future study of compounds 11 and 14 for their potential therapeutic use in conditions connected with collagen biosynthesis deficiency is required.

Precision-cut liver slices as a model for the early onset of liver fibrosis to test antifibrotic drugs

Induction of fibrosis during prolonged culture of precision-cut liver slices (PCLS) was reported. In this study, the use of rat PCLS was investigated to further characterize the mechanism of early onset of fibrosis in this model and the effects of antifibrotic compounds. Rat PCLS were incubated for 48h, viability was assessed by ATP and gene expression of PDGF-B and TGF-β1 and the fibrosis markers Hsp47, αSma and Pcol1A1 and collagen1 protein expressions were determined. The effects of the antifibrotic drugs imatinib, sorafenib and sunitinib, PDGF-pathway inhibitors, and perindopril, valproic acid, rosmarinic acid, tetrandrine and pirfenidone, TGFβ-pathway inhibitors, were determined. After 48h of incubation, viability of the PCLS was maintained and gene expression of PDGF-B was increased while TGF-β1 was not changed. Hsp47, αSma and Pcol1A1 gene expressions were significantly elevated in PCLS after 48h, which was further increased by PDGF-BB and TGF-β1. The increased gene expression of fibrosis markers was inhibited by all three PDGF-inhibitors, while TGFβ-inhibitors showed marginal effects. The protein expression of collagen 1 was inhibited by imatinib, perindopril, tetrandrine and pirfenidone. In conclusion, the increased gene expression of PDGF-B and the down-regulation of fibrosis markers by PDGF-pathway inhibitors, together with the absence of elevated TGF-β1 gene expression and the limited effect of the TGFβ-pathway inhibitors, indicated the predominance of the PDGF pathway in the early onset of fibrosis in PCLS. PCLS appear a useful model for research of the early onset of fibrosis and for testing of antifibrotic drugs acting on the PDGF pathway.

The molecular chaperone Hsp47 is essential for cartilage and endochondral bone formation

Heat shock protein 47 kDa (Hsp47) is considered as a molecular chaperone essential for the correct folding of type I and type IV procollagen in the ER. However, the function of Hsp47 for other types of procollagen and its importance for chondrogenesis have never been elucidated. To examine the function of Hsp47 in cartilage formation and endochondral ossification, we conditionally inactivated the Hsp47 gene in chondrocytes using Hsp47 floxed mice and mice carrying a chondrocyte-specific Col2a1-Cre transgene. Hsp47 conditional null mutant mice died just before or shortly after birth, and exhibited severe generalized chondrodysplasia and bone deformities with lower levels of type II and type XI collagen. Second-harmonic generation (SHG) analysis and electron microscopy revealed the accumulation of misaligned type I collagen molecules in the intervertebral discs and a substantial decrease in type II collagen fibers, respectively. Whole-mount skeletal staining showed no calcified region in the vertebral bodies of sacral vertebrae, and revealed that the endochondral bones were severely twisted and shortened. These results demonstrate that Hsp47 is indispensable for well-organized cartilage and normal endochondral bone formation.

IL-17 Promotes HSP47 Expression and Intestinal Fibrosis in Crohn's Disease

investigate the role of IL-17 in HSP47 expression and the potential for IL-17 as a new therapeutic target for intestinal fibrosis in CD. Methods: In Vivo study, serum levels of HSP47 in CD, UC and normal subjects were measured by enzyme-linked immunosorbent assay (ELISA). Localization of HSP47 positive cells in the intestine were evaluated by immunohistochemistry (IHC) using intestinal surgical specimens of patients with CD and UC. Gene expressions of HSP47, TGF-β-1, IL-13, HSP70 and IL-17 in the intestinal tissues were examined by quantitative real-time PCR (qRT-PCR). In Vitro study, gene expression of HSP47 in NIH3T3 cells after stimulation with TGF-β-1 or IL-17 was determined by qRT-PCR. Results: (1) Serum levels of HSP47 in patients with CD were significantly higher than in patients with UC and normal subjects. (2) IHC study revealed that HSP47 positive cells were mainly observed in mesenchymal site, submucosa and fibrostenotic site, and that the number of HSP47 positive cells were greater in the intestinal tissues of patients with CD than those with UC. (3) Gene expressions of HSP47, TGF-β-1 and IL-17 were significantly higher in the intestinal tissues of patients with CD than those with UC, while there were no significant differences of HSP70 and IL-13 gene expressions in the intestinal tissues between patients with CD and UC. (4) Gene expression of HSP47 in NIH3T3 cells were up-regulated by IL-17 stimulation as well as TGF-β-1. In addition, p38MAPK inhibitor suppressed HSP47 gene expression in NIH3T3 cells stimulated with IL-17. Conclusion: Our data suggest that IL-17 might be involved in intestinal fibrosis of CD by up-regulation of HSP47 expression in intestinal fibroblasts through p38MAPK pathway. IL-17 could be a new therapeutic target for not only intestinal inflammation but also intestinal fibrosis in CD.

炭酸ガスレーザー照射と象牙芽細胞の石灰化誘導

Reparative dentin is formed in the dental pulp in response to various external stimuli such as caries and abrasion. Carbon dioxide laser has been used to accelerate the formation of reparative dentin on the exposed pulp surface. However, the mechanism by which carbon dioxide laser irradiation stimulates mineralization in direct pulp capping treatment is not fully understood. We examined the effects of carbon dioxide laser irradiation on mineralization in rat dental pulp cells. Rat dental pulp cells were irradiated with a carbon dioxide laser at an output power of 2 W for 20, 40, and 60 s and were cultured in media containing ascorbic acid and β-glycerophosphate. Cell viability was examined 24 h after laser irradiation by a modified MTT assay. Alizarin Red S staining was performed 10 days after laser irradiation. The amounts of collagen secreted from the cells after irradiation were quantified following Sirius Red staining. The expression levels of collagen type I and HSP47, collagen-binding stress protein, were analyzed by real-time PCR. HSP47 protein expression was examined by Western blotting. The cell viability was not affected by laser irradiation at 2 W for up to 40 s. However, it was significantly decreased by 20% at 60 s. The amount of mineralization after 10 days of irradiation at 2 W for 40 s was significantly increased in comparison to the other conditions. The extracellular collagen production was significantly increased, by 73% on day 2 and 38% on day 4, after laser irradiation. Although collagen type I gene expression was not changed by laser irradiation, HSP47 gene and protein expression was induced within 12 h and 24 h, respectively. These results suggested that carbon dioxide laser irradiation stimulated mineralization in dental pulp cells by increasing HSP47 expression.

Carbon dioxide laser irradiation stimulates mineralization in rat dental pulp cells

To examine the effect of carbon dioxide laser irradiation on mineralization in dental pulp cells. Rat dental pulp cells were irradiated with a carbon dioxide laser at 2 W output power for 20, 40 and 60 s, and were cultured in ascorbic acid and beta-glycerophosphate containing media. Cell viability was examined 24 h after laser irradiation by a modified MTT assay. Alizarin Red S staining was performed 10 days after laser irradiation. The amounts of secreted collagen from the cells after irradiation were quantified following Sirius Red staining. The expression levels of collagen type I and HSP47, collagen-binding stress protein, were analysed by real-time PCR. HSP47 protein expression was examined by Western blotting. Statistical analysis was performed using one-way analysis of variance (anova) followed by the Tukey's multiple comparison test. The cell viability was not affected by laser irradiation at 2 W for up to 40 s. However, it was significantly decreased by 20% at 60 s (P < 0.05). The amount of mineralization after 10 days of irradiation at 2 W for 40 s was significantly increased in comparison to the other conditions (P < 0.05). The extracellular collagen production was significantly increased by 73% on day 2 and 38% on day 4 after laser irradiation (P < 0.05). Although collagen type I gene expression was not changed by laser irradiation, HSP47 gene and protein expression was induced within 12 and 24 h, respectively. These results suggested that carbon dioxide laser irradiation stimulated mineralization in dental pulp cells. The laser irradiation also increased HSP47 expression but not collagen gene expression.

The effect of recombinant HSP47siRNA on the pathological scar in nude mice: in vivo study

To study the significance of HSP47 gene in the development of pathological scar. The nude mice were used to reconstruct animal model of pathological scar. 16 days later, the mixture of recombinant HSP47siRNA and liposome was injected into the pathological scar in experimental group. In the control group, 0.25 ml PBS was injected intraperitoneally. 7 days after injection, the specimens were collected for detection of mRNA of HSP47, the collagen and for immunohistochemical study. In the control and experimental group, the collagen content was (91.71 +/- 1.24)% and (82.12 +/- 4.79)%, respectively; the expression of HSP47mRNA was 1 042 862.01 +/- 604 194.36 and 306 123.68 +/- 105 857.08, respectively; the expression of collagen I mRNA was 10 228 614.70 +/- 2 532 879.04 and 6 011 841.97 +/- 2 886 897.17, respectively;the scar volume was (255.60 +/- 21.34) mm3 and (132.99 +/- 24.06) mm3, respectively. All the above results showed significant difference between the two groups (P < 0.05). The collagen production can be reduced through suppression of the expression of HSP47 gene. It indicates that HSP47 gene enhance the development of keloid and could be used as the target of treatment.

Examination of the stress-induced expression of the collagen binding heat shock protein, hsp47, in Xenopus laevis cultured cells and embryos

HSP47 is an endoplasmic reticulum (ER)-resident molecular chaperone involved in collagen production. This study examined the stress-induced pattern of hsp47 gene expression in Xenopus cultured cells and embryos. Sequence analysis revealed that protein encoded by the hsp47 cDNA exhibited 70–77% identity with fish, avian and mammalian HSP47. In A6 kidney epithelial cells hsp47 mRNA and HSP47 were present constitutively and inducible by heat shock but not ER stressors including tunicamycin and A23187, both of which enhanced BiP mRNA. Furthermore A23187 treatment inhibited constitutive accumulation of hsp47 mRNA and retarded heat-induced accumulation of hsp47 and hsp70 mRNA. Interestingly, hsp47 gene expression but not hsp70 or BiP mRNA accumulation was enhanced by treatment with a procollagen-specific stressor, β-aminopropionitrile. In Xenopus embryos hsp47 mRNA was present constitutively throughout development. In tailbud embryos hsp47 mRNA was enriched in tissues associated with collagen production including notochord, somites and head region. Heat shock-induced accumulation of hsp47 mRNA was enhanced primarily in embryonic tissues already exhibiting hsp47 mRNA accumulation. These studies suggest that the pattern of Xenopus hsp47 gene expression is similar to hsp70 in response to heat shock but also displays unique features including a response to a procollagen-specific stressor and preferential expression in collagen-containing tissues.

Xantholipin, a Novel Inhibitor of HSP47 Gene Expression Produced by Streptomyces sp.

ChemInformVolume 34, Issue 43 Natural Products Xantholipin, a Novel Inhibitor of HSP47 Gene Expression Produced by Streptomyces sp. Yuichi Terui, Yuichi Terui Med. Res. Lab., Taisho Pharm. Co. Ltd., Omiya, Saitama 330, JapanSearch for more papers by this authoret al. et al., et al. et al. Med. Res. Lab., Taisho Pharm. Co. Ltd., Omiya, Saitama 330, JapanSearch for more papers by this author Yuichi Terui, Yuichi Terui Med. Res. Lab., Taisho Pharm. Co. Ltd., Omiya, Saitama 330, JapanSearch for more papers by this authoret al. et al., et al. et al. Med. Res. Lab., Taisho Pharm. Co. Ltd., Omiya, Saitama 330, JapanSearch for more papers by this author First published: 30 September 2003 https://doi.org/10.1002/chin.200343188Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article. Volume34, Issue43October 28, 2003 RelatedInformation