scholarly journals Structural Changes and Detection of Liver Fibrosis–Related Protein Levels in Coculture of Alveolar Echinococcosis-Protoscoleces and Human Hepatic Stellate Cells

2021 ◽  
Author(s):  
DEping cao ◽  
Emad Shamsan ◽  
Bofan Jiang ◽  
Zhang Yaogang ◽  
Mustafa Abdo Saif Dehwah
Keyword(s):  
Smooth Muscle ◽  
Liver Fibrosis ◽  
Hepatic Stellate Cells ◽  
Structural Changes ◽  
Alveolar Echinococcosis ◽  
Smooth Muscle Actin ◽  
Stellate Cells ◽  
Enzyme Linked Immunosorbent Assay ◽  
Muscle Actin ◽  
Related Protein

Abstract BackgroundEchinococcus multilocularis is a causative agent of human alveolar echinococcosis (AE). AE leads to cirrhosis in several organs, such as the liver, triggering severe conditions, including hepatic failure and encephalopathy. The main purpose of this study is to explore the interaction between treated hepatic stellate cells and AE-protoscoleces (AE-PSCs). The results of this study will be provided experimental basis for revealing the mechanisms of hepatic fibrosis after AE infection.MethodsWe investigated the role of alveolar echinococcosis-protoscoleces (AE-PSCs) in liver fibrosis and structural changes and liver fibrosis-related protein expression in a coculture of PSCs and human hepatic stellate cells (HSCs). Structural changes were detected by transmission electron microscopy, whereas liver fibrosis-related proteins, collagen I, alpha-smooth muscle actin, and osteopontin levels were measured by western blotting and enzyme-linked immunosorbent assay. ResultsPSCs exhibited morphological changes, specifically changes in shape, and showed slight changes in the cytoplasmic membrane, whereas structural modifications were observed in HSCs. Additionally, western blotting and enzyme-linked immunosorbent assay revealed that PSCs treated in vitro with HSC-LX2 showed significantly increased collagen-Ⅰ, α-smooth muscle actin, and osteopontin expression levels after 3–4 days of incubation in a coculture system. AE-PSCs induced liver fibrosis by inducing extracellular matrix expression and HSC activation.ConclusionsThese results provide insight into the pathogenesis of echinococcosis- induced hepatic fibrosis and introduce therapeutic targets and diagnostic criteria for managing echinococcosis-dependent liver fibrosis.

Dynamics of Sept4 expression in fibrotic livers of mice infected with Schistosoma japonicum

Parasitology ◽  
2011 ◽  
Vol 138 (8) ◽  
pp. 1003-1010 ◽  
Author(s):  
Y. N. DUAN ◽  
H. Y. QIAN ◽  
Y. W. QIN ◽  
D. D. ZHU ◽  
X. X. HE ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Liver Fibrosis ◽  
Mouse Model ◽  
Schistosoma Japonicum ◽  
Hepatic Stellate Cells ◽  
Immunohistochemical Staining ◽  
Smooth Muscle Actin ◽  
Stellate Cells ◽  
Muscle Actin ◽  
Qrt Pcr

SUMMARYIn order to investigate the dynamics of Septin4 (Sept4) expression and its function in the formation of fibrotic livers in mice infected with Schistosoma japonicum, we constructed the mouse model of S. japonicum egg-induced liver fibrosis for 24 weeks. Immunohistochemical staining, qRT-PCR and Western blot were used to detect the expression of Sept4 and α-smooth muscle actin (α-SMA). We found Sept4 localized in the perisinusoidal space where hepatic stellate cells (HSCs) distribute in the periphery of circumoval granulomas and the portal venule. The expression of Sept4 and α-SMA had a similar significant tendency of an up-regulation to a peak at 12 weeks post-infection (p.i.) followed by a down-regulation. At 24 weeks p.i. both were at a low level. These results suggest that Sept4 and α-SMA may interact together in HSCs. Based on this evidence, we hypothesize that Sept4 seems to be involved in the formation of inflammatory granulomata and subsequent liver fibrosis by regulating HSCs activation.

The hepatitis C virus core protein indirectly induces alpha-smooth muscle actin expression in hepatic stellate cells via interleukin-8

2010 ◽  
Vol 52 (5) ◽  
pp. 635-643 ◽  
Author(s):  
Sophie Clément ◽  
Stéphanie Pascarella ◽  
Stéphanie Conzelmann ◽  
Carmen Gonelle-Gispert ◽  
Kévin Guilloux ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Hepatitis C ◽  
Hepatic Stellate Cells ◽  
Core Protein ◽  
Smooth Muscle Actin ◽  
Stellate Cells ◽  
Muscle Actin ◽  
Alpha Smooth Muscle Actin ◽  
Virus Core ◽  
Actin Expression

scholarly journals Stellate cells in livers of dogs with portal vein hypoperfusion

2018 ◽  
Vol 74 (1) ◽  
pp. 6017-2018
Author(s):  
MAŁGORZATA SOBCZAK-FILIPIAK ◽  
JÓZEF SZAREK ◽  
MICHAŁ CZOPOWICZ ◽  
MAREK GALANTY ◽  
IZABELLA DOLKA ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Liver Fibrosis ◽  
Portal Vein ◽  
Lipid Droplets ◽  
Smooth Muscle Actin ◽  
Stellate Cells ◽  
Portosystemic Shunt ◽  
Muscle Actin ◽  
Healthy Dogs ◽  
Cytoplasmic Lipid Droplets

Hepatic stellate cells play a crucial role in the development of liver fibrosis. In a damaged liver, stellate cells undergo activation, which is manifested as a change of their phenotype: differentiation of stellate cells to myofibroblast-type cells, expression of alpha-Smooth Muscle Actin, their proliferation and a reduction in the size of cytoplasmic lipid droplets. The aim of this study was to determine the number and morphology of stellate cells in the canine liver affected by congenital portosystemic shunt (PSS) and portal vein hypoplasia – hepatic microvascular dysplasia (PVH-HMD). The material for investigation were archived paraffin blocks with liver samples collected supravitally from six dogs with PSS, six dogs with PVH-HMD and six healthy dogs. On the HE-stained sections, the number of stellate cells per 100 hepatocytes was counted (Sztark method) and the diameter of veins in the hepatic triads was measured (light microscope Olympus BX 43, SC30 camera, CellSens Entry 2011 Olympus). In addition, the diameter of lipid droplets in stellate cells was measured (computed image analysis system LUCIA 4.21). The results were analysed statistically (the Kruskal-Wallis test followed by Dunn’s post-hoc procedure; significance level (α) at 0.05; Statistica 12 StatSoft Inc.). The degree of liver fibrosis was determined (Masson’s method of slide stain; Scheuer scale). The liver samples from the dogs with PSS and PVH-HMD were stained immunohistochemically with Monoclonal Mouse Anti-Human Smooth Muscle Actin (α-SMA), clone 1A4, antibodies (DAKO). Portosystemic shunt and primary portal vein hypoplasia in the dog results in a reduction in the diameter of portal vein branches and in insufficient portal blood flow through the liver. In the material investigated, this was particularly evident in the animals affected by PSS: such dogs had a significantly smaller diameter of the veins than did the healthy dogs (p<0.001) or the dogs with PVH-HMD (p=0.023). Fibrosis and the expression of α-SMA were stronger in the dogs with PSS than in those with PVH-HMD. Moreover, the dogs with PSS had a significantly higher average number of stellate cells than the healthy animals (p=0.007) did. However, the examination of the material revealed an enlargement of cytoplasmic lipid droplets: the dogs with PSS had a significantly larger diameter of lipid vacuoles in the cytoplasm of stellate cells than did the healthy animals (p<0.001) or the dogs with PVH-HMD (p=0.043); the dogs with PVH-HMD had lipid droplets with a significantly larger diameter than the healthy animals (p<0.001) did. Hypoperfusion of the liver and the accompanying regressive lesions in hepatocytes result mainly in an increased number of stellate cells and stronger expression of α-SMA, while cytoplasmic lipid droplets in the stellate cells are not reduced in size. The present study indicates the need for detailed analyses of clinical cases and warrants further comprehensive studies of comparative hepatopathology because it demonstrates differences between humans and dogs in the morphological indicators of hepatic stellate cell activation in chronic liver damage.

c-Myb modulates transcription of the α-smooth muscle actin gene in activated hepatic stellate cells

2000 ◽  
Vol 278 (2) ◽  
pp. G321-G328 ◽  
Author(s):  
Martina Buck ◽  
Dong Joon Kim ◽  
Karl Houglum ◽  
Tarek Hassanein ◽  
Mario Chojkier
Keyword(s):  
Smooth Muscle ◽  
Hepatic Stellate Cells ◽  
Cell Activation ◽  
Stellate Cell ◽  
Smooth Muscle Actin ◽  
Stellate Cells ◽  
Dominant Negative ◽  
Muscle Actin ◽  
Muscle Actin Gene ◽  
High Level

Expression of α-smooth muscle actin (α-SMA) defines the phenotype of activated (myofibroblastic) hepatic stellate cells. These cells, but not quiescent stellate cells, have a high level of α-SMA and c-Myb expression, as well as increased c-Myb-binding activities to the proximal α-SMA E box. Therefore, we analyzed the role of c-Myb in α-SMA transcription and stellate cell activation. Activated primary rat stellate cells displayed a high expression of the −724 and −271 α-SMA/luciferase (LUC) chimeric genes, which contain c-Myb binding sites (−223/−216 bp). α-SMA/LUC minigenes with mutation (−219/−217 bp), truncation (−224 bp), or deletion (−191 bp) of the c-Myb binding site were not efficiently transcribed. Transfection of wild-type c-Myb into quiescent stellate cells, which do not express endogenous c-Myb, induced a ∼10-fold stimulation of −724 α-SMA/LUC expression. Conversely, expression of either a dominant-negative c-Myb basic domain mutant (Cys43 → Asp) or a c-Myb antisense RNA blocked transcription from the −724 α-SMA/LUC or −271 α-SMA/LUC in activated cells. Moreover, transfection of c- myb antisense, but not sense, RNA inhibited both expression of the endogenous α-SMA gene and stellate cell activation, whereas transfection of c- myb stimulated α-SMA expression in quiescent stellate cells. These findings suggest that c-Myb modulates the activation of stellate cells and that integrity of the redox sensor Cys43in c-Myb is required for this effect.

scholarly journals LincRNA-p21 Inhibits the Wnt/β-Catenin Pathway in Activated Hepatic Stellate Cells via Sponging MicroRNA-17-5p

2017 ◽  
Vol 41 (5) ◽  
pp. 1970-1980 ◽  
Author(s):  
Fujun Yu ◽  
Yong Guo ◽  
Bicheng Chen ◽  
Liang Shi ◽  
Peihong Dong ◽  
...  
Keyword(s):  
Liver Fibrosis ◽  
Hepatic Stellate Cells ◽  
Noncoding Rna ◽  
Type I Collagen ◽  
Smooth Muscle Actin ◽  
Stellate Cells ◽  
Type I ◽  
Muscle Actin ◽  
Pathway Activity ◽  
Long Intergenic Noncoding Rna

Background/Aims: It is known that the activation of hepatic stellate cells (HSCs) is a pivotal step in the initiation and progression of liver fibrosis. Aberrant activated Wnt/β-catenin pathway is known to accelerate the development of liver fibrosis. microRNAs (miRNAs)-mediated Wnt/β-catenin pathway has been reported to be involved in HSC activation during liver fibrosis. However, whether long noncoding RNAs (lncRNAs) regulate Wnt/β-catenin pathway during HSC activation still remains unclear. Methods: Long intergenic noncoding RNA-p21 (lincRNA-p21) expression was detected in Salvianolic acid B (Sal B)-treated cells. Effects of lincRNA-p21 knockdown on HSC activation and Wnt/β-catenin pathway activity were analyzed in Sal B-treated cells. In lincRNA-p21-overexpressing cells, effects of miR-17-5p on HSC activation and Wnt/β-catenin pathway activity were examined. Results: LincRNA-p21 expression was up-regulated in HSCs after Sal B treatment. In primary HSCs, lincRNA-p21 expression was down-regulated at Day 5 relative to Day 2. Sal B-inhibited HSC activation including the reduction of cell proliferation, α-smooth muscle actin (α-SMA) and type I collagen was inhibited by lincRNA-p21 knockdown. Sal B-induced Wnt/β-catenin pathway inactivation was blocked down by loss of lincRNA-p21. Notably, lincRNA-p21, confirmed as a target of miR-17-5p, suppresses miR-17-5p level. Lack of the miR-17-5p binding site in lincRNA-p21 prevents the suppression of miR-17-5p expression. In addition, the suppression of HSC activation and Wnt/β-catenin pathway induced by lincRNA-p21 overexpression was almost inhibited by miR-17-5p. Conclusion: We demonstrate that lincRNA-p21-inhibited Wnt/β-catenin pathway is involved in the effects of Sal B on HSC activation and lincRNA-p21 suppresses HSC activation, at least in part, via miR-17-5p-mediated-Wnt/β-catenin pathway.

scholarly journals De novo expression of glutamine synthetase during transformation of hepatic stellate cells into myofibroblast-like cells

1998 ◽  
Vol 335 (3) ◽  
pp. 697-700 ◽  
Author(s):  
Johannes G. BODE ◽  
Thorsten PETERS-REGEHR ◽  
Axel M. GRESSNER ◽  
Dieter HÄUSSINGER
Keyword(s):  
Enzyme Activity ◽  
Smooth Muscle ◽  
Glutamine Synthetase ◽  
Western Blot ◽  
Hepatic Stellate Cells ◽  
De Novo ◽  
Smooth Muscle Actin ◽  
Stellate Cells ◽  
Cell Protein ◽  
Muscle Actin

The expression of glutamine synthetase (GS) was studied in cultured quiescent hepatic stellate cells (HSC) and during their transformation into myofibroblast-like cells. GS mRNA was detectable in quiescent HSC (1-day culture); however, the enzyme protein was not expressed, as assessed by Western blot analysis, immunocytochemistry and the absence of detectable enzyme activity. Similar findings were obtained after 2 days of culture; in addition, the mRNA levels had dropped by about 70%, but they increased again thereafter during the process of HSC transformation in culture, as indicated by the expression of α-smooth-muscle actin. In parallel with the accumulation of α-smooth-muscle actin, GS was expressed, as shown by Western blot analysis and immunocytochemistry, and enzyme activity increased from undetectable levels in quiescent cells to 0.13±0.01 µmol/h per mg of cell protein within 7–14 days. This value compares with GS activity in liver parenchymal cells of 0.57±0.03 µmol/h per mg of cell protein. The findings suggest that activation of HSC results in the de novo expression of GS protein and activity, and this may serve as another marker of HSC transformation.

Increased contractility of hepatic stellate cells in cirrhosis is mediated by enhanced Ca2+-dependent and Ca2+-sensitization pathways

2011 ◽  
Vol 300 (6) ◽  
pp. G1010-G1021 ◽  
Author(s):  
Masateru Iizuka ◽  
Takahisa Murata ◽  
Masatoshi Hori ◽  
Hiroshi Ozaki
Keyword(s):  
Smooth Muscle ◽  
Hepatic Stellate Cells ◽  
Smooth Muscle Actin ◽  
Stellate Cells ◽  
Regulatory Proteins ◽  
Muscle Actin ◽  
Rock Inhibitor ◽  
Duct Ligation

Activation of hepatic stellate cells (HSCs) results in cirrhosis and portal hypertension due to intrahepatic resistance. Activated HSCs increase their contraction after receptor agonist stimulation; however, the signaling pathways for the regulation of contraction are not fully understood. The aim of this study was to elucidate the change in contractile mechanisms of HSCs after cirrhotic activation. The expression pattern of contractile regulatory proteins was analyzed with quantitative RT-PCR and Western blotting. The phosphorylation levels of myosin light chain (MLC), 17-kDa PKC-potentiated protein phosphatase 1 inhibitor protein (CPI-17), and MLC phosphatase targeting subunit 1 (MYPT1) after endothelin-1 (ET-1) stimulation in culture-activated HSCs were measured using phosphorylation-specific antibodies. In vivo-activated HSCs were isolated from rats subjected to bile duct ligation and repeated dimethylnitrosoamine injections. HSCs showed increased expression of not only α-smooth muscle actin, but also the contractile regulatory proteins MLC kinase (MLCK), Rho kinase 2 (ROCK2), and CPI-17 during HSC activation in vitro. In culture-activated HSCs, ET-1 increased phosphorylation of CPI-17 at Thr18, which was markedly inhibited by the PKC inhibitor Ro-31–8425. ET-1 induced phosphorylation of MYPT1 at Thr853, which was suppressed by the ROCK inhibitor Y-27632. ET-1 induced sustained phosphorylation of MLC at Thr18/Ser19, which was inhibited by both Ro-31–8425 and Y-27632. Consistent with the data obtained from the in vitro study, HSCs isolated from cirrhotic rats showed increased expression of α-smooth muscle actin, MLCK, CPI-17, and ROCK2 compared with HSCs from nontreated rats. Furthermore, MLC phosphorylation in in vivo-activated HSCs was increased, according to enhanced phosphorylation of CPI-17 and MYPT1 in the presence of ET-1. These results suggest that activated HSCs may participate in constriction of hepatic sinusoids in the cirrhotic liver through both Ca2+-dependent (MLCK pathway) and Ca2+-sensitization mechanism (CPI-17 and MYPT1 pathways).

Can hepatic stellate cells express alpha-smooth muscle actin in normal human liver?

2001 ◽  
Vol 21 (4) ◽  
pp. 293-294 ◽  
Author(s):  
Sébastien Lepreux ◽  
Liliane Dubuisson ◽  
Brigitte Le Bail ◽  
Alexis Desmoulière ◽  
Charles Balabaud ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Hepatic Stellate Cells ◽  
Human Liver ◽  
Smooth Muscle Actin ◽  
Stellate Cells ◽  
Muscle Actin ◽  
Alpha Smooth Muscle Actin ◽  
Normal Human Liver ◽  
Normal Human
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