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Effects of 17β-Estradiol on the Plasminogen Activator System in Vascular Smooth Muscle Cells Treated with Lysophophatidylcholine

Yoon BK1,2Kang YH3Oh WJ3Lee DY1Kim DK4Kessel B5Kang CD6.

J Menopausal Med. 2020 Apr;26(1):9-17. doi: 10.6118/jmm.19005.

 

Abstract

OBJECTIVES:

When administered soon after menopause, hormone therapy can prevent coronary heart diseases in women. To explore the mechanism underlying the cardioprotective actions of estrogen, we investigated the effects of 17β-estradiol (17β-E₂) on the plasminogen activator system using cultured vascular smooth muscle cells (VSMCs).

METHODS:

VSMCs were isolated from rat aortas. Protein expression of plasminogen activator inhibitor type 1 (PAI-1) and tissue-type plasminogen activator (t-PA) were evaluated using Western blotting and enzyme-linked immunosorbent assay, respectively. The enzyme activity of PAI-1 in a conditioned medium was assessed via reverse fibrin overlay zymography and that of t-PA was assessed via fibrin overlay zymography. Gene expression was quantified using real-time reverse transcription-polymerase chain reaction.

RESULTS:

Following pre-treatment for 24 hours, 17β-E₂ suppressed both protein expression and enzyme activity of PAI-1 stimulated by lysophosphatidylcholine (lysoPC) in a significant and dose-dependent manner at a near physiological concentration. Moreover, 17β-E₂ (10⁻⁷ M) inhibited PAI-1 gene expression, and ICI 182,780-a specific estrogen receptor antagonist-blocked the effects of 17β-E₂ on the PAI-1 protein. 17β-E₂ did not affect t-PA secretion but significantly enhanced free t-PA activity through reduced binding to PAI-1. Furthermore, 17β-E₂ suppressed intracellular reactive oxygen species production and nuclear factor-κB-mediated transcription.

CONCLUSIONS:

In VSMCs stimulated with lysoPC, 17β-E₂ reduced PAI-1 expression through a non-receptor-mediated mechanism via antioxidant activity as well as a receptor-mediated mechanism; however, it did not alter t-PA secretion. Of note, 17β-E₂ suppressed PAI-1 activity and concurrently enhanced t-PA activity, suggesting a beneficial influence on fibrinolysis.