Angiotensin II induces contraction and proliferation of human hepatic stellate cells

doi:10.1016/S0016-5085(00)70368-4

Gastroenterology

Volume 118, Issue 6, June 2000, Pages 1149-1156

https://doi.org/10.1016/S0016-5085(00)70368-4 Get rights and content

Abstract

Background & Aims: Circulating levels of angiotensin II (ANGII), a powerful vasoconstrictor factor, are frequently increased in chronic liver diseases. In these conditions, hepatic stellate cells (HSCs) proliferate and acquire contractile properties. This study investigated the presence of receptors for ANGII and the effects of ANGII in human HSCs activated in culture. Methods: The presence of ANGII receptors was assessed by binding studies. The effects of ANGII on intracellular calcium concentration ([Ca²⁺]_i), cell contraction, and cell proliferation were also assessed. Results: Binding studies showed the presence of ANGII receptors of the AT1 subtype. ANGII elicited a marked dose-dependent increase in [Ca²⁺]_i and cell contraction. Moreover, ANGII stimulated DNA synthesis and increased cell number. All these effects were totally blocked by losartan and reduced by nitric oxide donors or prostaglandin E₂. The effects of ANGII were barely detectable in quiescent cells (2 days in culture), suggesting that phenotypic transformation of HSCs is associated with a marked increase in the effects of ANGII. Conclusions: ANGII induces contraction and is mitogenic for human-activated HSCs by acting through AT1 receptors. These results suggest that activated HSCs are targets of the vasoconstrictor action of ANGII in the intrahepatic circulation.

GASTROENTEROLOGY 2000;118:1149-1156

Section snippets

Isolation and culture of human and rat HSCs

Human HSCs were isolated from fragments of normal liver tissue obtained from patients with liver metastases undergoing liver resection (n = 4) and from a liver explant of a patient with a metabolic disease undergoing liver transplantation (n = 1). Methods of cell isolation have been described in detail previously.⁶ Nutritional status was normal in all patients. No patient had ascites or edema or had fluid restriction before the surgical procedure. Liver function test results were normal in all

Receptor binding

The binding of [¹²⁵I]ANGII to activated human HSCs was time and temperature dependent (data not shown). Maximal specific binding was obtained after 60 minutes at 22.25°C. [¹²⁵I]ANGII specifically bound to human HSCs with a K_d of 0.25 ± 0.03 nmol/L and a B_max of 13.6 ± 0.7 fmol/well. The number of ANGII binding sites per cell was 19,804 ± 630 (mean number of cells/well, 4.14 ± 0.07 × 10⁵) (Figure 1A).

. (A) Scatchard plot of [¹²⁵I]ANGII binding studies to human activated HSCs. K_d, 0.25 nmol/L; B_max

Discussion

This study shows that human HSCs activated in culture have receptors for ANGII that mediate cell contraction and proliferation, effects similar to those reported for VSMCs and mesangial cells.12, 19, 21, 22 Two different types of ANGII receptors have been described. The AT1 receptors are present in most mesenchymal cells and mediate most of the biological effects of ANGII, including the increase in [Ca²⁺]_i, cell contraction, and proliferation.18, 23 These effects can be blocked by specific AT1

Acknowledgements

The authors thank Esther Tobías for excellent technical assistance and Drs. Josep Fuster, Juan Carlos García-Valdecasas, Luis Grande, and Antonio Lacy for providing liver tissues for isolation of HSCs. Losartan and PD123,319 were a kind gift from Merck Sharpe & Dohme and Research Biochemicals International, respectively.

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^☆: Address Requests for reprints to: Pere Ginès, M.D., Liver Unit, Hospital Clínic, Villarroel 170, 08036-Barcelona, Spain. Fax: (34) 93-451-52-72.

^☆☆: Supported by a grant from the Dirección General de Enseñanza Superior e Investigación Técnica (SAF99/0014) and the Secretaría de Estado para Universidades e Investigación (PM96-0058), a fellowship grant from the Consell Interdepartamental de Recerca I Innovació Tecnológica (CIRIT) (to R.B.), and a grant from the Fundació Clínic per a la Recerca Biomèdica (to M.N.G. and E.G.-R.).

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Liver, Pancreas, and Biliary TractAngiotensin II induces contraction and proliferation of human hepatic stellate cells☆,☆☆

Abstract

Section snippets

Isolation and culture of human and rat HSCs

Receptor binding

Discussion

Acknowledgements

Hepatology

Gastroenterology

Hepatology

Kidney Int

Hepatology

J Hepatol

Gastroenterology

J Hepatol

Desmin and actin in the identification of Ito cells and in monitoring their evolution to myofibroblasts in experimental liver fibrosis

Virchows Arch B Cell Pathol

Activation-dependent contractility of rat hepatic lipocytes in culture and in vivo

J Clin Invest

The contraction of hepatic stellate (Ito) cells stimulated with vasoactive substances. Possible involvement of endothelin-1 and nitric oxide in the regulation of the sinusoidal tonum

Eur J Biochem

Fat-storing cells as liver-specific pericytes. Spatial dynamics of agonist-stimulated intracellular calcium transients

J Clin Invest

Renin-angiotensin-aldosterone system and the renal regulation of sodium, potassium, and blood pressure homeostasis

Cellular mechanisms mediating rat renal microvascular constriction by angiotensin II

J Clin Invest

Characterization of angiotensin II receptor subtypes in human glomeruli and mesangial cells

Am J Physiol

Glomerular mesangial cells: electro-physiology and regulation of contraction

Physiol Rev

Effect of drug infusion on the splanchnic circulation. I. Angiotensin infusion in normal and cirrhotic subjects

Proc Soc Exp Biol Med

Effect of angiotensin-II blockade on systemic and hepatic hemodynamics and on the renin-angiotensin-aldosterone system in cirrhosis with ascites

Eur J Clin Invest

Liver, Pancreas, and Biliary Tract
Angiotensin II induces contraction and proliferation of human hepatic stellate cells☆,☆☆