Elsevier

Pathophysiology

Volume 11, Issue 3, December 2004, Pages 153-158
Pathophysiology

Mechanical stretch induces matrix metalloproteinase 1 production in human hepatic stellate cells

https://doi.org/10.1016/j.pathophys.2004.07.003Get rights and content

Abstract

In increasing portal blood flow, hepatic stellate cells (HSC) may be lengthened in response to mechanical stretch stimulation and their function may be changed. However, little is known about the influence of mechanical stretch on hepatic stellate cells. We examined production of matrix metalloproteinases (MMP), tissue inhibitors of metalloproteinases (TIMP), and extracellular matrix by hepatic stellate cells to investigate the relationship between mechanical stretch and hepatic fibrosis. LI90 cells, human hepatic stellate cells, were stretched cyclically using the Flexer cell strain unit. Concentrations of MMP1, MMP2, TIMP1, TIMP2, type I collagen C-telopeptide (1CTP), procollagen III propeptide (PIIIP), and hyaluronic acid in culture supernatants were determined. MMP1, MMP2, and TIMP1 mRNA expression was measured by reverse transcription-polymerase chain reaction (RT-PCR). In stretched LI90 cells, concentration of MMP1 showed an increase relative to unstretched cells, but concentrations of MMP2, TIMPl, and TIMP2 showed a decrease. MMP1/TIMP1 ratio and MMP1 mRNA expression showed an increase in stretched cells. Our finding suggested that in the early phase of portal hypertension, hepatic stellate cells increase production of MMPl and decrease production of TIMP1 and TIMP2, activated by mechanical stretch.

Introduction

Hepatic stellate cells (HSC), which correspond to pericytes surrounding capillaries, are located within the subendothelial space of Disse, are characterized by long, branching processes and take part in the pathogenesis of liver disease [1], [2]. Hepatic stellate cells store retinyl palmitate, deliver retinol to extrahepatic tissues, regulate sinusoidal microcirculation, synthesize extracellular matrix proteins, and participate in fibrotic scarring in chronic hepatic diseases.

In chronic hepatic diseases, especially liver cirrhosis, portal hypertension is one of the most important complications. The pressure gradient within the portal venous system is a product of vascular resistance (backward flow theory) [3], [4] and portal blood flow (forward flow theory) [5], [6]. Increasing portal blood flow, HSC may be lengthened in response to mechanical stretch stimulation and their function may be changed.

Hepatic fibrosis results from a relative imbalance between synthesis and degradation of matrix proteins. In liver injury, both the amount and relative composition of the hepatic extracellular matrix are altered. Matrix metalloproteinases (MMP) have a function in the turnover of extracellular matrix components such as collagens, proteoglycans, elastin, laminin, fibronectin, and other glycoproteins, while tissue inhibitors of metalloproteinases (TIMP) are low-molecular weight proteins that inhibit MMP in a 1:1 molar ratio [7], [8]. Iredale et al. reported that TIMP1 expression increased relative to MMP1 and TIMP1 expression and preceded procollagen 1 expression in liver after bile duct ligation and carbon tetrachloride administration [9]. Herbst et al. reported that HSC expressed increased encoding of TIMP1 and TIMP2 mRNA in liver fibrosis and in vitro, TGF-β1 enhanced expression of TIMP1 and MMP2 mRNA [10]. HSC, then, produce MMP and TIMP, shifting their balance in many kinds of liver injury.

MMP1, also termed interstitial collagenase, degrades collagens I, II, III, VII, and X as well as gelatins, entactin, aggrecan, and link protein. MMP2, or gelatinase A, degrades gelatins, collagens I, IV, V, VII, X, and XI, fibronectin, laminin, aggrecan, elastin, large tenascin C, and vitronectin. MMP2 also degrades β-amyloid protein precursor, thus showing β-secretase-like activity [8]. Type I and type III collagen accounts for about 85% of all extracellular matrix protein in the liver. Okazaki et al. reported that collagenase activity increased and MMP1 played an important role in experimental hepatic fibrosis [11].

Previous studies have revealed a relationship between mechanical stretch and matrix metalloproteinases and tissue inhibitors of metalloproteinases in many kinds of cultured cells. However, little is known about the influence of mechanical stretch stimulation on production of MMP and TIMP in HSC, which play an important role in hepatic fibrosis. Therefore, we examined production of MMP, TIMP, and extracellular matrix by HSC to investigate the relationship between mechanical stretch and hepatic fibrosis.

Section snippets

Cell culture

We used a cultured human hepatic stellate cell line, LI90 (provided by Dr. Tomokazu Matsuura), in this study. This cell line, established by Murakami et al. from an epithelioid hemangioendothelioma arising in human liver, is well characterized as consisting of HSC-like cells that lack characteristics of endothelial cells or macrophages [12]. LI90 cells had a polygonal shape, had well developed α-smooth muscle actin filaments in their cytoplasm, and produced various connective tissue components,

Morphological change of stretched LI90 cells

In phase-contrast microscopic studies, LI90 cells stretched for 24 h were narrower than unstretched cells and showed a finer network of processes (Fig. 1).

Concentration of MMP1, MMP2, TIMP1, and TIMP2 in culture supernatants of LI90 cells

The concentration of MMP1 in culture supernatants showed an increase in stretched as opposed to unstretched LI90 cells, while in contrast the concentrations of MMP2, TIMP1, and TIMP2 showed a decrease in stretched cells compared to unstretched cells: MMP1 in unstretched cells, 197.8 ± 23.0 ng/ml, and stretched cells, 224.8 ± 13.9 ng/ml (p < 0.05);

Discussion

Portal hypertension, which results from increased intrahepatic resistance to blood flow after liver injury, is a major complication of scarring in the liver. Von Leeuwen et al. reported Disse's space was significantly increased in patients with chronic active hepatitis (CAH), CAH in transition to cirrhosis, and cirrhosis compared with near normal subjects [15]. In the hyperdynamic portal circulation and increase Disse's space, HSC may be exposed to mechanical stress caused by congested blood

References (21)

There are more references available in the full text version of this article.

Cited by (27)

  • Lipid based nanocarriers for effective drug delivery and treatment of diabetes associated liver fibrosis

    2021, Advanced Drug Delivery Reviews
    Citation Excerpt :

    MMPs are the enzymes associated with ECM degradation and TIMP1 has MMP inhibiting activity. Hence, it is essential to maintain balanced MMP-TIMP levels for efficient ECM remodeling [35]. Among several types of MMPs like MMP-1, MMP-2, MMP-8, MMP-9, MMP-10; MMP-9 has a central role in ECM homeostasis [36].

  • Anesthetic Considerations for Fontan-Associated Liver Disease and the Failing Fontan Circuit

    2020, Journal of Cardiothoracic and Vascular Anesthesia
    Citation Excerpt :

    The study investigators concluded that this subgroup of patients had considerable liver fibrosis, most likely present even before completion of the Fontan procedure, and that the associated portal fibrosis in their series probably reflected the multifactorial etiology of liver fibrosis in the SV patient population.48 One possible mechanism of hepatic fibrosis in pre-Fontan patients, even at the stage 2 Glenn surgery, is hepatic venous blood flow reversal caused by atrial contraction, leading to intrahepatic reflux, associated hepatic vein distention, stromal stretch, and compression of adjacent hepatocytes.24,49 All patients with the Fontan circulation require regular screening of the liver with age-appropriate laboratory investigations and imaging.9

  • Combined hepatocellular-cholangiocarcinoma after tetralogy of Fallot repair: a case report and review of literature

    2020, Pathology Research and Practice
    Citation Excerpt :

    In their review of several studies, Rockey et al. wrote that the mechanisms responsible for fibrogenesis are likely to involve several inducers released from hepatic stellate cells [15]. Goto et al. showed that the stimuli from the stretching and compression of hepatic stellate cells may be a potential inducer of liver fibrosis [16]. In a murine model, Simonetto et al. demonstrated that hepatic congestion led to sinusoidal thrombosis and strain, which in turn promoted hepatic fibrosis via the release of fibronectin hepatic stellate cells [17].

  • Multiscale computational model of fluid flow and matrix deformation in decellularized liver

    2016, Journal of the Mechanical Behavior of Biomedical Materials
    Citation Excerpt :

    Lobule displacements yielded matrix strain predictions ranging from 1.7% to 6.1% (Sections S.2, S.3). To put these values in context, hepatic stellate cells have exhibited gene expression changes with application of cyclic 10% strain (Takashi et al., 2004). Thus, the ~5% strain observed in the present model is in the range of biologically meaningful mechanical stretch.

  • Molecular pathogenesis of hepatic fibrosis and current therapeutic approaches

    2011, Chemico-Biological Interactions
    Citation Excerpt :

    Among the cells involved in hepatic ECM degradation are HSC, neutrophils, and macrophages. MMPs are the main enzymes responsible for ECM degradation and TIMPs have the ability to inhibit MMPs [70]. Therefore, regulation of the MMP-TIMP balance is crucial for efficient ECM remodeling.

View all citing articles on Scopus
View full text